tag:blogger.com,1999:blog-2047344909668062522024-03-27T16:53:32.361-07:00 ARVIGEN HEALTHCAREGeneration towards Healthcare Bringing Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.comBlogger54125tag:blogger.com,1999:blog-204734490966806252.post-59528134254037988842022-09-05T10:51:00.002-07:002022-09-05T10:53:45.913-07:00Study Shows MVT-602 Be a Novel Fertility-Improving Drug<p> </p><h1 class="title" itemprop="headline" style="background-color: white; box-sizing: border-box; font-family: Inter, "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 2.1875rem; letter-spacing: -0.05em; line-height: 2.8125rem; margin: 20px 0px; padding-bottom: 30px; text-align: center; text-rendering: optimizelegibility;">MVT-602 is a Novel Fertility-Improving Drug</h1><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhbb_75SaPNpPzdofQ16lgMvE7XiLxyqo-AWISBK78eCXZq501VX4Cha12p080Qy4szXOmbB6qGZ8UkliBiskdlYZaAA4vECYnNrTguCWFv6esiW_cscZC1LFx0G3xK3CJJcYVkBp1IXpyyn49MYcz_gTFwTGX2z5xB6M3q8XINl6Rcoka64l7z2uWPkg/s646/images%20(2).jpeg" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="474" data-original-width="646" height="235" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhbb_75SaPNpPzdofQ16lgMvE7XiLxyqo-AWISBK78eCXZq501VX4Cha12p080Qy4szXOmbB6qGZ8UkliBiskdlYZaAA4vECYnNrTguCWFv6esiW_cscZC1LFx0G3xK3CJJcYVkBp1IXpyyn49MYcz_gTFwTGX2z5xB6M3q8XINl6Rcoka64l7z2uWPkg/s320/images%20(2).jpeg" width="320" /></a></div><br /><div><br /></div><div><p style="background-color: white; box-sizing: border-box; color: #606267; font-family: Inter, "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 16px; letter-spacing: -0.04em; line-height: 1.5625rem; margin: 0px 0px 25px;">FERTILITY in women with reproductive health problems can be improved using a new drug according to a recent study from researchers and clinicians at Imperial College London, London, UK.</p><p style="background-color: white; box-sizing: border-box; color: #606267; font-family: Inter, "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 16px; letter-spacing: -0.04em; line-height: 1.5625rem; margin: 0px 0px 25px;">Lead author Prof Waljit Dhillo noted that: “Reproductive health issues are common for women around the world. Infertility as a result of these conditions can cause a lot of distress.” Kisspeptin-54 (KP54), a naturally occurring form of kisspeptin, has been researched for several years to treat reproductive disorders. Previous studies had shown that kisspeptin can be used to safely stimulate reproductive hormones in women undergoing <em style="box-sizing: border-box;">in vitro</em> fertilisation treatment without causing ovarian hyperstimulation syndrome. However, in this study, the researchers investigated whether MVT-602, a novel drug that targets the kisspeptin system to stimulate reproductive hormones that affect fertility, menstruation, and sexual development, could target the kisspeptin pathway and produce a longer hormonal release than KP54.</p><p style="background-color: white; box-sizing: border-box; color: #606267; font-family: Inter, "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 16px; letter-spacing: -0.04em; line-height: 1.5625rem; margin: 0px 0px 25px;">The study was conducted from 2017 to 2019 and included 24 females aged 18–35 years who were given MVT-602; 12 were healthy volunteers and 12 were patients with polycystic ovary syndrome or hypothalamic amenorrhea. The healthy volunteers were given an injection of the KP54 and saline placebo for comparison. First, the reproductive hormone levels after receiving MVT-602 and KP54 were compared, and then the reproductive hormone levels after MVT-602 between healthy patients, patients with hypothalamic amenorrhea, and those with polycystic ovary syndrome were compared.</p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEj1PZ9DNkMzQ8BQJdrBQYGEn63PSPAQ9yUElo4tlXbaKhvMKiwkePngsydiMkUbOtH2n6FzHjhWhlU2M1gmeojY3pva84nOUWFZTGtTEH7upF07mSYFupPAUa-9NyjhhPEMsK_kXujwV4OH3AaQ5uOjx1ZPeyvskphpNPao2CSOyMleB9btlO5M3SpRiw/s678/images%20(1).jpeg" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="452" data-original-width="678" height="213" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEj1PZ9DNkMzQ8BQJdrBQYGEn63PSPAQ9yUElo4tlXbaKhvMKiwkePngsydiMkUbOtH2n6FzHjhWhlU2M1gmeojY3pva84nOUWFZTGtTEH7upF07mSYFupPAUa-9NyjhhPEMsK_kXujwV4OH3AaQ5uOjx1ZPeyvskphpNPao2CSOyMleB9btlO5M3SpRiw/s320/images%20(1).jpeg" width="320" /></a></div><br /><p style="background-color: white; box-sizing: border-box; color: #606267; font-family: Inter, "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 16px; letter-spacing: -0.04em; line-height: 1.5625rem; margin: 0px 0px 25px;"><br /></p><p style="background-color: white; box-sizing: border-box; color: #606267; font-family: Inter, "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 16px; letter-spacing: -0.04em; line-height: 1.5625rem; margin: 0px 0px 25px;">Results highlighted that patients given MVT-602 showed a longer duration of raised luteinising hormone and follicle stimulating hormone levels than when they received KP54. Fellow study author Dr Ali Abbara emphasised that: “This is the first study to show that a single dose of MVT-602 can induce a longer duration of hormonal stimulation in women than naturally occurring kisspeptin. Therefore, it reveals exciting potential to treat a range of reproductive health conditions using MVT-602 and offers women improved treatment options. However, further research is needed to fully characterise its effects in specific disorders that affect reproductive health.” The authors are hopeful that in the future, MVT-602 could be used to treat a wider range of reproductive disorders.</p></div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-49151227656971830082022-07-10T10:32:00.003-07:002022-07-10T10:32:39.098-07:00There is a cycle to cycle variation in ovarian response and pre-hCG serum progesterone level: an analysis of 244 consecutive IVF cycles<h1 class="c-article-title" data-article-title="" data-test="article-title" style="background-color: white; box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1.875rem; letter-spacing: min(max(-0.0117156rem, 4vw), -0.0390625rem); line-height: 1.2; margin: 0px 0px 16px; padding: 0px;">There is a cycle to cycle variation in ovarian response and pre-hCG serum progesterone level: an analysis of 244 consecutive IVF cycles</h1><ul class="c-article-author-list js-etal-collapsed js-no-scroll" data-component-authors-activator="authors-list" data-etal-small="3" data-etal="25" data-test="authors-list" style="background-color: white; box-sizing: inherit; color: #222222; display: inline; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; list-style: none; margin: 0px 8px 0px 0px; padding: 0px; width: 100%;"><li class="c-article-author-list__item" style="box-sizing: inherit; display: inline; margin-left: 0px; padding-right: 0px;">Sule Yildiz, </li><li class="c-article-author-list__item" style="box-sizing: inherit; display: inline; margin-left: 0px; padding-right: 0px;">Kayhan Yakin, </li><li class="c-article-author-list__item js-smaller-author-etal" style="box-sizing: inherit; display: inline; margin-left: 0px; padding-right: 0px;">Baris Ata & </li><li class="c-article-author-list__item" style="box-sizing: inherit; display: inline; margin-left: 0px; padding-right: 0px;">Ozgur Oktem<svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-email" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg> </li></ul><span style="background-color: white; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 18px;"></span><p class="c-article-info-details" data-container-section="info" style="background-color: white; box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin: 16px 0px 8px; overflow-wrap: break-word; padding: 0px; word-break: break-word;"><i data-test="journal-title" style="box-sizing: inherit;">Scientific Reports</i> <span data-test="journal-volume" style="box-sizing: inherit; font-weight: bolder;"><span class="u-visually-hidden" style="border: 0px; box-sizing: inherit; clip: rect(0px, 0px, 0px, 0px); height: 1px; margin: -1663px; overflow: hidden; padding: 0px; position: absolute !important; width: 1px;">volume</span> 10</span>, Article number: <span data-test="article-number" style="box-sizing: inherit;">15793</span> (<span data-test="article-publication-year" style="box-sizing: inherit;">2020</span>) Cite this article</p><div class="c-article-metrics-bar__wrapper u-clear-both" style="background-color: white; 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margin-right: 1em;"><img border="0" data-original-height="525" data-original-width="700" height="240" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEii3A9gJoQStS2LoDI8n03qdtZXCudvZleTZjrLJ2Ywl7UWMDesioK_qCg7386fBtDPKfhZbNedvqrVvC9GVJOg_jnkv4gGqWM3R43zIkoZNVCmyAnjfS613XRYy-AmfHnvBcUbbMtcAGyWDLwL4IVrwsPlRr5kkYFsJEgRB6bJRXxu6nKT4SYBfsMWVw/s320/navbharat-times.webp" width="320" /></a></div><br /><div><br /></div><div><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Abs1" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Abstract</h2><div class="c-article-section__content" id="Abs1-content" style="box-sizing: inherit; font-size: 18px; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">We aimed to answer one key question, that was not previously addressed as to whether serum progesterone (P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4-hCG</span> day) and its co-variates (estradiol (E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2-hCG day</span>) and the number of retrieved oocytes) of a given cycle can be predictive of the subsequent cycle when both cycles are consecutive and comparable for the stimulation protocol, gonadotropin dose and duration of stimulation. We analyzed such 244 consecutive (< 6 months) IVF cycles in 122 patients with GnRH agonist long protocol and found that P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span>, E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> and the number of retrieved oocytes significantly vary between the two cycles. Although P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> increased (ranging from 4.7 to 266.7%) in the 2nd cycle in 61 patients, E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> and the number of retrieved oocytes, which are normally positively correlated with P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> paradoxically decreased in the 41% and 37.7% respectively, of these same 61 patients. When a similar analysis was done in the 54 out of 122 patients (44.3%) in whom serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> was decreased in the 2nd cycle, the mean decrease in P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> was − 34.1 ± 23.3% ranging from − 5.26 to − 90.1%. E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> and the number of retrieved oocytes paradoxically increased in the 42.3% and 40.7% of these 54 patients respectively. P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> remained the same only in the 7 (5.7%) of these 122 patients. These findings indicate that late follicular phase serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> may change unpredictably in the subsequent IVF cycle. The changes are not always necessarily proportional with ovarian response of previous cycle suggesting that growth characteristics and steroidogenic activities of antral cohorts may exhibit considerable cycle to cycle variations.</p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjp3MYzvRfFFlpxigjZTHhQjZ1Jf6XGCG-dIqQIX6R89LAhY4XmannYrzLQ3RIOabSgVog0AEk6vA_hIsf499Z8NhVZOvOkK7fb4in24DqyvAiP_rqX7yWIC4U8u69SKgA_hMq1RpzsdpA5V0XZKqAUh0WimxvVsxhHhkMCyGxtOApvPtgTmkOnzILapA/s1280/maxresdefault.jpg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="720" data-original-width="1280" height="180" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjp3MYzvRfFFlpxigjZTHhQjZ1Jf6XGCG-dIqQIX6R89LAhY4XmannYrzLQ3RIOabSgVog0AEk6vA_hIsf499Z8NhVZOvOkK7fb4in24DqyvAiP_rqX7yWIC4U8u69SKgA_hMq1RpzsdpA5V0XZKqAUh0WimxvVsxhHhkMCyGxtOApvPtgTmkOnzILapA/s320/maxresdefault.jpg" width="320" /></a></div><br /><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;"><br /></p><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec1" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Introduction</h2><div class="c-article-section__content" id="Sec1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Serum progesterone (P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span>) level may elevate during late follicular phase of ovarian stimulation before ovulation is triggered and is considered a negative predictive factor for clinical outcome in assisted reproduction in both GnRH agonist and GnRH antagonist protocols<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 1" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#ref-CR1" id="ref-link-section-d165748016e374" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Venetis, C. A., Kolibianakis, E. M., Bosdou, J. K. & Tarlatzis, B. C. Progesterone elevation and probability of pregnancy after IVF: a systematic review and meta-analysis of over 60 000 cycles. Hum. Reprod. Update 19, 433–457.
https://doi.org/10.1093/humupd/dmt014
(2013).">1</a></span>. It is not a true luteinization event because the elevations in serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level occurs prior to hCG administration and is not associated with any premature LH surge. This phenomenon is generally observed in stimulated IVF cycles and shows significant correlation with the intensity of ovarian stimulation; hence patients with more growing follicles and retrieved oocytes have higher P levels. Interestingly, it was also documented that pre-ovulatory P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> elevations may also occur in up to 28% of natural cycles and reduce pregnancy rates. However, the mechanism of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> rise in natural cycles is distinct from stimulated IVF cycles because there is no gonadotropin stimulation or multiple follicle development in the former. In agreement with the close link between the degree of ovarian stimulation with gonadotropins and serum pre-hCG P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level in stimulated IVF cycles, we recently provided a molecular explanation for this phenomenon by showing that FSH stimulation itself promotes P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> synthesis and output from human granulosa cells without luteinization by up-regulating the expression and enzymatic activity of the enzyme 3β-hydroxy steroid dehydrogenase (3β-HSD), which converts pregnenolone to progesterone. Therefore, it is likely that pre-ovulatory P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> elevation is caused by the insufficiency of the ovary in handling the increased output of precursor steroids generated during multi-follicular development in FSH stimulated IVF cycles. To date, several clinical studies and meta-analyses showed a negative impact of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> elevations before ovulation trigger on the chance of pregnancy in fresh embryo transfer IVF cycles</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">In most of the cases in stimulated IVF cycles, the risk for an elevation in serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level at late follicular phase before ovulation trigger is closely related to the magnitude of ovarian stimulation, that is also reflected by E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> level on the hCG day, and the number of growing follicles > 14 mm on day 10 of stimulation and total and mature oocytes retrieved. Therefore, there are significant positive associations between P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> and these co-variates. However, one key question remains to be answered as to whether we should expect to see similar pre-hCG serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> levels and its co-variates in two consecutive IVF cycles comparable for gonadotropins, GnRH analog used, and duration of stimulation. We aimed to answer this question in this non-interventional, retrospective cohort data from a single center.</p><section data-gtm-vis-first-on-screen-50443292_562="174405" data-gtm-vis-first-on-screen-50443292_563="174405" data-gtm-vis-has-fired-50443292_562="1" data-gtm-vis-has-fired-50443292_563="1" data-gtm-vis-total-visible-time-50443292_562="10000" data-gtm-vis-total-visible-time-50443292_563="10000" data-title="Material and methods" style="box-sizing: inherit;"><div class="c-article-section" id="Sec2-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec2" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Material and methods</h2><div class="c-article-section__content" id="Sec2-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><h3 class="c-article__sub-heading" id="Sec3" style="box-sizing: inherit; font-size: 1.5rem; font-weight: 400; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Study design</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">This study is a retrospective cohort analysis from a single center. It was approved by the Institutional Review Board (IRB) of Koc University (2015.206.IRB2.076). It is not a research study that involve human participation. Therefore, the need of the written informed consent was waived by the Institutional Review Board (IRB) of Koc University. All methods were performed in accordance with the relevant guidelines and regulations of the Institution. We reviewed the electronic database of 8724 IVF cycles that had been performed from 2008 to 2015 in Assisted Reproduction Unit, American Hospital of Istanbul, Turkey. A total of 122 women who had 244 consecutive IVF cycles within a 6-month interval following an unsuccessful cycle, using exactly the same ovarian stimulation protocol in both cycles were identified and included in this study. Patients who underwent stimulation more than 6 months apart or had ovarian surgery, systemic disease that could affect ovarian response to stimulation were excluded.</p><h3 class="c-article__sub-heading" id="Sec4" style="box-sizing: inherit; font-size: 1.5rem; font-weight: 400; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Ovarian stimulation and ovulation trigger</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Pituitary down-regulation was induced with GnRH agonist leuprolide acetate started 7 days prior to the anticipated day of menstrual bleeding and continued until the day of hCG trigger. Recombinant FSH was started on cycle day three at a dose of 225–300 IU depending upon age, antral follicle count, anticipated or documented previous ovarian response, and body mass index. Ovulation was triggered with 250 µg recombinant hCG (Ovitrelle; Merck-Serono, Istanbul, Turkey) when a leading follicle of ≥ 19 mm and two or more trailing follicles of ≥ 17 mm were recorded. Follicular aspiration was performed 36 h after ovulation trigger. Oocyte retrieval was performed under general anaesthesia using a double lumen needle (Cook Ireland Ltd., Limerick, Ireland) as we described previously.</p><h3 class="c-article__sub-heading" id="Sec5" style="box-sizing: inherit; font-size: 1.5rem; font-weight: 400; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Hormone assays</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Serum samples for hormone assays were obtained by veni-puncture and assessed using a validated electrochemiluminescence immunoassay (ECLIA method, Cobas 6000, Roche, Basel, Switzerland) as described previously<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 11" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#ref-CR11" id="ref-link-section-d165748016e472" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Oktem, O. et al. High responders are not exempt from detrimental effects of prematurely rising progesterone levels in fresh embryo transfer cycles. Reprod. Biomed. Online 38, 206–215.
https://doi.org/10.1016/j.rbmo.2018.11.008
(2019).">11</a></span>. Analytical sensitivity (lower detection limit) for P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> was 0.095 nmol/L (0.030 ng/mL) and the functional sensitivity (defined as lowest analyte concentration that can be reproducibly measured with a between-run coefficient of variation [CV] of < 20%) was 0.48 nmol/L (0.15 ng/mL). The day-to-day CV was 2.9% at 2.31 nmol/L (0.73 ng/mL), 1.4% at 9.57 nmol/L (3.1 ng/mL), and 0.9% at 103 nmol/L (32.4 ng/mL). Analytical sensitivity for E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> was 18.4 pmol/L (5 pg/mL). The day-to-day CV for E2 was 6.7% at 27.4 pg/mL, 1.1% at 1270 pg/mL, and 1.9% at 2720 pg/ml. The same assay was used during the study period and was calibrated whenever a new reactive batch was used or whenever an outcome outside the normal range was observed.</p><h3 class="c-article__sub-heading" id="Sec6" style="box-sizing: inherit; font-size: 1.5rem; font-weight: 400; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Statistical analysis</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Continuous variables in the baseline demographic and IVF characteristics were expressed as mean (SD) or median (25<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">th</span>–75th percentile) depending on distribution characteristics. Two-tailed Pearson correlation test and linear regression analysis were used to identify the confounding variables that show significant association with serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level. Continuous variables were compared between the groups with paired samples t- test or Wilcoxon signed rank test as appropriate. The significance level was set at 5% (P < 0.05). Graphpad Prism (version 7) and SPSS statistical programs (version 23) were used to analyse the data and create the figures<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 11" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#ref-CR11" id="ref-link-section-d165748016e492" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Oktem, O. et al. High responders are not exempt from detrimental effects of prematurely rising progesterone levels in fresh embryo transfer cycles. Reprod. Biomed. Online 38, 206–215.
https://doi.org/10.1016/j.rbmo.2018.11.008
(2019).">11</a></span>.</p></div></div></section><section data-gtm-vis-first-on-screen-50443292_562="213899" data-gtm-vis-first-on-screen-50443292_563="213899" data-gtm-vis-total-visible-time-50443292_562="6000" data-gtm-vis-total-visible-time-50443292_563="6000" data-title="Results" style="box-sizing: inherit;"><div class="c-article-section" id="Sec7-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec7" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Results</h2><div class="c-article-section__content" id="Sec7-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Baseline demographic and IVF characteristics of the two consecutive cycles are summarized in Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#Tab1" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;">1</a>. Male factor infertility (78.5%) was the major indication followed by unexplained infertility (9.4%), tubal factor (11.9), and ovulatory pathologies (1.6%). Average age, daily and total doses of gonadotropins and duration of stimulation, serum levels of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> and E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> levels on the hCG day, the numbers of fol > 14 mm on day 10 of stimulation and total and mature oocytes retrieved were similar between the two cycles (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#Tab1" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;">1</a>). In both cycles, serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level on the hCG day was significantly associated with serum E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> level on the hCG day, and the numbers of growing follicles (fol > 14 mm) on day 10 of stimulation, and the oocytes retrieved on the correlation and linear regression analyses (Fig. 1). However, the level of significance was not the same for all three co-variates of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span>. E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> on the hCG day and total oocyte counts appeared to be more closely associated with P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> in both 1st and 2nd IVF cycles than the number of fol > 14 mm.</p><div class="c-article-table" data-container-section="table" data-test="inline-table" id="table-1" style="border: 2px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption class="c-article-table__figcaption" style="box-sizing: inherit; line-height: 1.4; margin-bottom: 16px; word-break: break-word;"><span data-test="table-caption" id="Tab1" style="box-sizing: inherit; font-weight: bolder;">Table 1 Baseline demographic and IVF characteristics of the two consecutive IVF cycles.</span></figcaption><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-gtm-vis-first-on-screen-50443292_561="214080" data-gtm-vis-total-visible-time-50443292_561="1200" data-test="figure" data-title="Figure 1" id="figure-1" style="border: 2px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig1" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Figure 1</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0/figures/1" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-72597-0/MediaObjects/41598_2020_72597_Fig1_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure 1" aria-describedby="Fig1" height="242" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-72597-0/MediaObjects/41598_2020_72597_Fig1_HTML.png" style="border: 0px; box-sizing: inherit; display: block; height: auto; margin: 0px auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-1-desc" style="box-sizing: inherit; font-size: 1rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">The association of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level on the hCG day with E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">2</span> level on the hCG day, and the numbers of fol > 14 mm and retrieved oocytes on the correlation and linear regression analyses in the 1st and 2nd IVF cycles.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">The mean values of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> and its co-variates (serum E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> level on the hCG day, and the numbers of growing follicles (fol > 14 mm) on day 10 of stimulation, and the oocytes retrieved) were comparable between the cycles. However, we noticed there were substantial cycle to cycle variations in these markers (Fig. 2).</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-gtm-vis-first-on-screen-50443292_561="214294" data-gtm-vis-total-visible-time-50443292_561="1700" data-test="figure" data-title="Figure 2" id="figure-2" style="border: 2px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig2" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Figure 2</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0/figures/2" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-72597-0/MediaObjects/41598_2020_72597_Fig2_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure 2" aria-describedby="Fig2" height="411" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-72597-0/MediaObjects/41598_2020_72597_Fig2_HTML.png" style="border: 0px; box-sizing: inherit; display: block; height: auto; margin: 0px auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-2-desc" style="box-sizing: inherit; font-size: 1rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Comparison of the means of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">4</span> on the hCG day (<span style="box-sizing: inherit; font-weight: bolder;">A</span>) and its co-variates E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">2</span> on the hCG day (<span style="box-sizing: inherit; font-weight: bolder;">B</span>), and the numbers of fol > 14 mm on day 10 of stimulation (<span style="box-sizing: inherit; font-weight: bolder;">C</span>) and retrieved oocytes (<span style="box-sizing: inherit; font-weight: bolder;">D</span>) between the 1st and 2nd IVF cycles are shown as scatter plot with bars (the left images graphics). The variations in these parameters between the cycles are also shown as lines </p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Next, we investigated how serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level and its correlates changed in the 2nd IVF cycle in comparison to their corresponding 1st cycle values in each patient. This was expressed as the percentage of change based on the formula as follows: [(P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4 in the 2nd cyle </span>− P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4 in the 1st cyle</span>) × 100/P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4 in the 1st cyle</span>)]. We observed that there are indeed significant variations in the 2nd cycle when compared to their corresponding 1st cycle values of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> (%change: − 90.1 to 266.7%), E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> (− 81.7 to 424%), Fol > 14 mm (− 80 to 180%) and oocyte number (− 80 to 200%). The mean, median and percentiles of the percent changes for each of these variables are shown in the Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#Tab2" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;">2</a> and depicted as a scatter plot diagram in the Fig. 3A. Then, the IVF cycles were categorized into three groups according to the percent change of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> in the second IVF cycle as being > 0; = 0 and < 0 in order to investigate how the co-variates of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> (E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2-hCG day</span>, fol > 14 mm and retrieved oocyte counts) changed in relation to a particular change in P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span>. By doing so we aimed to identify the cycles in which the co-variates of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> exhibited paradoxical rather than parallel changes with P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4.</span> in the subsequent cycles in comparison to the first cycles.</p><div class="c-article-table" data-container-section="table" data-test="inline-table" id="table-2" style="border: 2px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption class="c-article-table__figcaption" style="box-sizing: inherit; line-height: 1.4; margin-bottom: 16px; word-break: break-word;"><span data-test="table-caption" id="Tab2" style="box-sizing: inherit; font-weight: bolder;">Table 2 Descriptive summary statistics of the percent changes of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> on the hCG day and its correlates E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> on the hCG day, and the numbers of fol > 14 mm on day 10 of stimulation and retrieved oocytes.</span></figcaption><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-gtm-vis-first-on-screen-50443292_561="215871" data-gtm-vis-total-visible-time-50443292_561="2400" data-test="figure" data-title="Figure 3" id="figure-3" style="border: 2px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig3" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Figure 3</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0/figures/3" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-72597-0/MediaObjects/41598_2020_72597_Fig3_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure 3" aria-describedby="Fig3" height="549" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-72597-0/MediaObjects/41598_2020_72597_Fig3_HTML.png" style="border: 0px; box-sizing: inherit; display: block; height: auto; margin: 0px auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-3-desc" style="box-sizing: inherit; font-size: 1rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Scatter plot diagrams of the percent changes of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">4</span> on the hCG day and its correlates E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">2</span> on the hCG day, and the numbers of fol > 14 mm on day 10 of stimulation and retrieved oocytes overall (<span style="box-sizing: inherit; font-weight: bolder;">A</span>), and when P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">4</span> in the second cycle is > 0 (<span style="box-sizing: inherit; font-weight: bolder;">B</span>), < 0 (<span style="box-sizing: inherit; font-weight: bolder;">C</span>) or = 0 (<span style="box-sizing: inherit; font-weight: bolder;">D</span>). Paradoxical changes in the co-variates of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">4</span> (Y-axis) when P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">4</span> increased and decreased are visible on the X-axis.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">According to the categorization described above serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level on the hCG day was higher in the 2nd IVF cycle than their corresponding 1st cycle levels in 61 of 122 patients (50.0%); remained unchanged in the seven (5.7%) and decreased in the 54 patients (44.3%). The mean increase in P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> in the 2nd cycle was 50.6 ± 53.3% ranging from 4.76 to 266.6%. However, the mean and range of changes in the co-variates of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> were 20.3% (− 81.8 to 424.6%) for E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span>; 10.9% (− 61.9 to 150%) for fol > 14 mm; and 11.2% (− 58.3 to 200%) for total oocyte numbers, suggesting that not all elevations in P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> are accompanied by parallel increases in its-covariates in the 2nd IVF cycle (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#Tab3" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;">3</a>, Fig. <a data-track-action="figure anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#Fig3" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;">3</a>B). We found that E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> and the numbers of fol > 14 mm and retrieved oocytes paradoxically decreased in the 55%, 37.7% and 41% respectively, of these 61 patients despite the elevations in P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> levels in their 2nd IVF cycle. The percent decreases ranged from − 2.03 to − 81.8 for E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span>; from − 6.67 to − 61.9 for Fol > 14 mm; and from − 7.14 to − 58.3 for the oocyte count. The distribution of the parallel and paradoxical changes in the co-variates of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> in relation to the changes in P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> are depicted as a histogram in the Fig. 4</p><div class="c-article-table" data-container-section="table" data-test="inline-table" id="table-3" style="border: 2px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption class="c-article-table__figcaption" style="box-sizing: inherit; line-height: 1.4; margin-bottom: 16px; word-break: break-word;"><span data-test="table-caption" id="Tab3" style="box-sizing: inherit; font-weight: bolder;">Table 3 Descriptive summary statistics of the percent changes of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> in the 2nd cycle > 0.</span></figcaption><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-gtm-vis-first-on-screen-50443292_561="218133" data-gtm-vis-total-visible-time-50443292_561="1000" data-test="figure" data-title="Figure 4" id="figure-4" style="border: 2px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig4" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Figure 4</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0/figures/4" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-72597-0/MediaObjects/41598_2020_72597_Fig4_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure 4" aria-describedby="Fig4" height="144" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-72597-0/MediaObjects/41598_2020_72597_Fig4_HTML.png" style="border: 0px; box-sizing: inherit; display: block; height: auto; margin: 0px auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-4-desc" style="box-sizing: inherit; font-size: 1rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Histogram depiction of the percent change of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">4</span> in the 2nd cycle (x-axis) and the corresponding percent change in E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">2</span>, and the numbers of fol > 14 mm and retrieved oocytes (y-axis). Light green areas show the cycles in which there are parallel changes in P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">4</span> and its co-variates (E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">2</span> and the numbers of fol > 14 mm and retrieved oocytes). Light blue areas demonstrate the cycles in which there were paradoxical changes in P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">4</span> and its correlates. Solid line: linear regression, dotted line 95% confidence intervals.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">When a similar analysis was done in the 54 out of 122 patients (44.3%) in whom serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> was decreased in the 2nd cycle, the mean decrease in P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> was − 34.1 ± 23.3% ranging from − 5.26 to − 90.1% (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#Tab4" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;">4</a>). It appeared that E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> and the numbers of fol > 14 mm and retrieved oocytes paradoxically increased in the 42.3%, 45.3% and 40.7% of these 54 patients respectively. The percent increases ranged from 1.60 to 136.3% for E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span>; from 6.25 to 180 for fol > 14 mm; and from 8.3 to 100 for the oocyte count (Figs. 3C and 4).</p><div class="c-article-table" data-container-section="table" data-test="inline-table" id="table-4" style="border: 2px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption class="c-article-table__figcaption" style="box-sizing: inherit; line-height: 1.4; margin-bottom: 16px; word-break: break-word;"><span data-test="table-caption" id="Tab4" style="box-sizing: inherit; font-weight: bolder;">Table 4 Descriptive summary statistics of the percent changes of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> in the 2nd cycle < 0.</span></figcaption><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Since the number of patients was too small (n = 7) in the group where serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level remain unchanged between the cycles no statistical analysis was conducted (Fig. <a data-track-action="figure anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#Fig3" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;">3</a>D).</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">We also investigated if P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> elevation in the previous cycle can predict its occurrence in the next cycle because it was shown that history of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> elevation can predict its occurrence again in the next cycle independent of ovarian stimulation. When a cutoff level of 1.5 ng/mL was adopted for serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> we found that 57 (46.7%) of 122 patients had serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level ≥ 1.5 ng/mL in the 1st cycle. And 32 (56.1%) of these 57 patients continued to have P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level exceeding this threshold in the 2nd cycle. Interestingly, despite P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> elevations, 14 (43.8%) of these 32 women had paradoxical decreases in the number of retrieved oocytes in their 2nd cycle with the percent decreases ranging from − 7.14 to − 80.0. Similar decreases (− 2.03 to − 65.2%) were observed in the E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> level in 21 of these 32 patients (67.7%).</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Most of the patient in this cohort 94/122 (77%) were normal responders (4–15 oocytes in the 1st cycle). Therefore we conducted a subgroup analysis analysis for this group and obtained similar results: Serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level on the hCG day in the 2nd IVF cycle was higher than their corresponding 1st cycle levels in 48 of 94 patients (51.1%), while it remained unchanged in five (5.3%) and decreased in the 54 patients (43.6%). E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> and the numbers of fol > 14 mm and retrieved oocytes paradoxically decreased in the 51.1%, 35.4% and 33.3% of these 48 patients respectively. The percent decreases ranged from − 2.03 to − 81.8 for E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span>; from − 6.67 to − 57.1 for Fol > 14 mm; and from − 7.14 to − 58.3 for the oocyte count (Supplementary data).</p></div></div></section><section data-gtm-vis-first-on-screen-50443292_562="219428" data-gtm-vis-first-on-screen-50443292_563="219428" data-gtm-vis-total-visible-time-50443292_562="4700" data-gtm-vis-total-visible-time-50443292_563="4700" data-title="Discussion" style="box-sizing: inherit;"><div class="c-article-section" id="Sec8-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec8" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Discussion</h2><div class="c-article-section__content" id="Sec8-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">We have shown in this study that serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level on the hCG day and its co-variates (E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> on the hCG day and the numbers of growing follicles > 14 mm on day 10 of stimulation and retrieved oocytes) may exhibit significant cycle to cycle variations even if both cycles are consecutive and comparable for the stimulation protocol, the type and dose of gonadotropin and duration of stimulation. The increase in serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level in the 2nd cycle was not always associated with parallel increases in its co-variates. There were some paradoxical inverse changes in these co-variates that were normally supposed to be in a positive association with P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span>. Even though P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> was significantly associated with E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> and the numbers of fol > 14 mm and retrieved oocytes in each cycle itself, not all decrease or elevations in P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> in the subsequent cycle were accompanied by parallel changes in its co-variates. These findings suggest that the growth and steroidogenic characteristics of antral cohorts in response to exogenous FSH may vary and may not reliably be predictive of the next cycle even if both cycles are comparable and successive.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">It is unclear why there are differences between two cohort of antral follicles of two different cycles exposed to FSH at the same dose and duration. Previous studies documented that there might be fluctuations in the number of antral follicles and AMH levels at early follicular phase between the cycles. Inter-cycle variability might not be confined to cohort of growing antral follicles itself as there could be differences in the expression of FSH receptors of granulosa cells of growing follicles and their response to exogenous FSH. Intraovarian actions of FSH and/or the degree of ovarian stimulation might be responsible for premature P output from granulosa cells without luteinization. Steroidogenic activity of the ovarian follicle changes depending on this developmental stage as well as its receptor abundancy and sensitivity. Ovarian stimulation with exogenous FSH is a continuum of multifollicular development with a number of follicles at different stages of development all contributing to the total steroid synthesis at different levels, any given time-point, such as the day of ovulation triggering. Even when the level of steroidogenesis in each granulosa cell or growing follicle does not increase, total steroid synthesis would increase as a factor of increased number of growing follicles and their steroidogenic granulosa cell mass. Another plausible explanation would be the increase in number or sensitivity of FSH and/or LH receptors on the granulosa cells in response to exogenous gonadotropin stimulation. If there are intrinsic differences among the cohorts of antral follicles recruited by FSH in terms of their FSH receptor expression and responsiveness, their growth kinetics and steroidogenic activity may change from cycle to cycle. A particular P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level at late follicular phase that was reached after achieving a certain magnitude of ovarian response to stimulation by FSH in one IVF cycle might not necessarily produce the same degree of ovarian stimulation and P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> levels in the next cycle. Variations in the level of significance for the co-variates of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> between the cycles supports this notion while recognizing at the same time that these differences could be related to the relatively small number of subjects in each cycle.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">We have recently provided molecular evidence that FSH itself up-regulates the expression and enzymatic activity of the enzyme 3β-hydroxy steroid dehydrogenase (3β-HSD) and promotes P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> output from human granulosa cells and ovarian tissue samples without luteinization<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 9" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#ref-CR9" id="ref-link-section-d165748016e2088" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Oktem, O. et al. FSH Stimulation promotes progesterone synthesis and output from human granulosa cells without luteinization. Hum. Reprod.
https://doi.org/10.1093/humrep/dex010
(2017).">9</a></span>. Therefore, serum P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level before ovulation trigger is significantly associated with the markers of the degree of ovarian stimulation in multivariate regression analysis, that are the co-variates of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> and include the number of growing follicles in response to FSH on day 10 of stimulation, E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> level on the hCG day and the numbers of oocytes retrieved.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Data regarding variations in the hormonal profile of women undergoing a similar type ovarian stimulation is limited. A retrospective analysis of 197 women with multiple treatment cycles, showed that premature progesterone elevation is likely to recur in repetitive stimulation cycles (OR 8.4; 95% CI 2.8–24.9). The increased likelihood of recurrence persisted when the regression model was adjusted for the intensity of ovarian stimulation. In the same study, basal P level at the initiation of stimulation was independently associated with P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> levels on triggering day. Authors suggested that in the presence of a corpus luteum that had not undergone functional luteolysis might be responsible from high levels of late follicular phase P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span>. It was also suggested that persistent high P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> synthesis in repeated cycles might be related with patient-specific intrinsic defects in ovarian or adrenal steroidogenesis<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 18" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-72597-0#ref-CR18" id="ref-link-section-d165748016e2116" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="De Geyter, C., De Geyter, M., Huber, P. R., Nieschlag, E. & Holzgreve, W. Progesterone serum levels during the follicular phase of the menstrual cycle originate from the crosstalk between the ovaries and the adrenal cortex. Hum. Reprod. 17, 933–939.
https://doi.org/10.1093/humrep/17.4.933
(2002).">18</a></span>. In our study, 32 of 122 women (26.2%) had persistent high (≥ 1.5) P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> levels in both cycles. Interestingly, 14 (43.8%) of these women had paradoxical decreases in the number of retrieved oocytes in their 2nd cycle with the percent decreases ranging from − 7.14 to − 80.0. Similar paradoxical decreases (− 2.03 to − 65.2%) were observed in the E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> level on the hCG day of the 2nd IVF cycle in 21 of these 32 patients (67.7%). These results provide additional evidence that the co-variates of P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> may not reliably predict its elevation again in the subsequent cycle. A recent prospective study reported significant intra-day variations in serum progesterone levels during the day of ovulation trigger (Gonzales-Foruria et al. 2019). P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> level was highest early in the morning and then gradually decreased throughout the day. Hormone measurements in the blood samples were performed at the same time period of the day early in the morning between 08:00 and 10:00 AM in our study.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">These findings were obtained in a relatively small number of IVF patients most of whom were good responders and male factor infertility was the main indication for IVF in majority of the cases. Currently, it is unclear if these results vary depending upon infertility etiology and the types of ovarian stimulation protocol and ovarian response. This is also true when there are accompanying ovarian pathology or other disease that may alter ovarian response to stimulation such as endometriosis, which is a complex disease with genetic, epigenetic and immunologic aberrations. Diminished ovarian reserve or poor response to stimulation are commonly observed in patients with endometriosis undergoing ovulation induction or ovarian stimulation. Despite many efforts we still do not have a serum hormone marker or a correct algorithm to choose the optimal starting dose of FSH in patients with low and high ovarian reserve and in those with PCOS and high AMH. Apparently, bi-directional communication between the oocyte and cumulus granulosa cells plays role in the response to gonadotropins, ovulation, oocyte maturation and IVF success. It is unknown if this bi-directional communication varies from follicle to follicle in a given cycle or between the two consecutive cycles. It also should be remembered that all required steps of controlled ovarian stimulation should be accomplished for best practice in IVF as there are other key factors in addition to premature P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> elevation that might impact the success of IVF such as embryo transfer technique.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">To the best of our knowledge this is the first study that analyzed such characteristics of two successive IVF cycles. However, there are several imitations of this study such as its retrospective design, data from a single center, and inclusion of a highly specific group of patients with two consecutive ovarian stimulations cycles within a specified time-period using exactly the same stimulation protocol. Although the design was intended to limit confounding variables like changes in stimulation protocol, dose of gonadotropins and ovarian aging, it limits the number of patients in the study and compromises the generalizability of the findings.</p></div></div></section><section data-gtm-vis-first-on-screen-50443292_562="223425" data-gtm-vis-first-on-screen-50443292_563="223425" data-gtm-vis-total-visible-time-50443292_562="2900" data-gtm-vis-total-visible-time-50443292_563="2900" data-title="Conclusion" style="box-sizing: inherit;"><div class="c-article-section" id="Sec9-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec9" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Conclusion</h2><div class="c-article-section__content" id="Sec9-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;">Serum progesterone (P<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4-hCG</span> day) and its co-variates (estradiol (E<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2-hCG day</span>) and the numbers of growing follicles > 14 mm and retrieved oocytes) may exhibit significant variations between the two cycles even when both cycles are consecutive and comparable for the stimulation protocol, gonadotropin dose and duration of stimulation. Therefore, the growth characteristics and steroidogenic activities of growing antral cohorts might change from cycle to cycle. The parameters of a previous IVF cycle might not accurately predict the subsequent cycle. These findings need to be confirmed in larger number of IVF patients with different infertility etiologies, ovarian stimulation protocols and ovarian response types.</p></div></div></section><div id="MagazineFulltextArticleBodySuffix" style="box-sizing: inherit; margin: 0px; padding: 0px;"><section aria-labelledby="Bib1" data-gtm-vis-first-on-screen-50443292_563="223659" data-gtm-vis-has-fired-50443292_563="1" data-gtm-vis-recent-on-screen-50443292_563="223659" data-gtm-vis-total-visible-time-50443292_563="10000" data-title="References" style="box-sizing: inherit;"><div class="c-article-section" id="Bib1-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Bib1" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.0117156rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">References</h2><div class="c-article-section__content" id="Bib1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><div data-container-section="references" style="box-sizing: inherit; margin: 0px; padding: 0px;"><ol class="c-article-references" style="box-sizing: inherit; list-style: none; margin-bottom: 28px; margin-top: 0px; padding: 0px;"><li class="c-article-references__item js-c-reading-companion-references-item" data-counter="1." style="border-bottom: 1px solid rgb(213, 213, 213); box-sizing: inherit; display: flex; flex-wrap: wrap; margin-bottom: 16px; padding-bottom: 16px;"><p class="c-article-references__text" id="ref-CR1" style="box-sizing: inherit; flex: 1 1 0%; margin: 0px 0px 8px; overflow-wrap: break-word; padding: 0px 0px 0px 8px; word-break: break-word;">Venetis, C. 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box-sizing: inherit; color: #006699; overflow-wrap: break-word; padding-left: 8px; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;">Google Scholar</a> </p></li></ol></div></div></div></section></div></div><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; word-break: break-word;"><br /></p></div></div></div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com1tag:blogger.com,1999:blog-204734490966806252.post-54217137559850477102021-12-19T21:31:00.004-08:002021-12-19T21:46:53.260-08:00The effect of Propofol versus Dexmedetomidine as anesthetic agents for oocyte pick-up on in vitro fertilization (IVF) outcomes<p> </p><li class="c-article-author-list__item" style="box-sizing: inherit; color: #222222; display: inline; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin-left: 0px; padding-right: 0px;"><div></div></li><li class="c-article-author-list__item" style="box-sizing: inherit; color: #222222; display: inline; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin-left: 0px; padding-right: 0px;"><br /><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/a/AVvXsEhmQl8CX7SiBRNvCKiV8bR0TOcwFqE7tVqwIwsCD4Zv8h63Bspo7i-PRYiTbFUTai47qdYSUshLCTLOzEKEBOwIjNDrpXWflAl6sZKMIaYTZvvH6Xha5DM83Dq6zUZXn207TJ0_ja0z7q-FqsoWg1OfVmeweDWwJd5kmETnkrNC59Vhz0Cvqb_Cu04_Ag=s824" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="518" data-original-width="824" height="331" src="https://blogger.googleusercontent.com/img/a/AVvXsEhmQl8CX7SiBRNvCKiV8bR0TOcwFqE7tVqwIwsCD4Zv8h63Bspo7i-PRYiTbFUTai47qdYSUshLCTLOzEKEBOwIjNDrpXWflAl6sZKMIaYTZvvH6Xha5DM83Dq6zUZXn207TJ0_ja0z7q-FqsoWg1OfVmeweDWwJd5kmETnkrNC59Vhz0Cvqb_Cu04_Ag=w643-h331" width="643" /></a></div><br /></li><div><li class="c-article-author-list__item" style="box-sizing: inherit; color: #222222; display: inline; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin-left: 0px; padding-right: 0px;"><br /></li></div>Özcan Budak, <li class="c-article-author-list__item" style="box-sizing: inherit; color: #222222; display: inline; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin-left: 0px; padding-right: 0px;">Mehmet Sühha Bostancı<svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-email" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg>, </li><li class="c-article-author-list__item js-smaller-author-etal" style="box-sizing: inherit; color: #222222; display: inline; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin-left: 0px; padding-right: 0px;">AyçaTaş Tuna, </li><li class="c-article-author-list__item js-smaller-author-etal" style="box-sizing: inherit; color: #222222; display: inline; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin-left: 0px; padding-right: 0px;">Veysel Toprak, </li><li class="c-article-author-list__item js-smaller-author-etal" style="box-sizing: inherit; color: #222222; display: inline; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin-left: 0px; padding-right: 0px;">Hüseyin Çakiroğlu & </li><li class="c-article-author-list__item" style="box-sizing: inherit; color: #222222; display: inline; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin-left: 0px; padding-right: 0px;">Koray Gök </li><span face="-apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif" style="background-color: white; color: #222222; font-size: 18px;"></span><p class="c-article-info-details" data-container-section="info" style="background-color: white; box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin: 16px 0px 8px; padding: 0px;"><i data-test="journal-title" style="box-sizing: inherit;">Scientific Reports</i> <span data-test="journal-volume" style="box-sizing: inherit; font-weight: bolder;"><span class="u-visually-hidden" style="border: 0px; box-sizing: inherit; clip: rect(0px, 0px, 0px, 0px); height: 1px; margin: -1663px; overflow: hidden; padding: 0px; position: absolute; width: 1px;">volume</span> 11</span>, Article number: <span data-test="article-number" style="box-sizing: inherit;">23922</span> (<span data-test="article-publication-year" style="box-sizing: inherit;">2021</span>) </p><section aria-labelledby="Abs1" data-gtm-vis-first-on-screen-50443292_562="523" data-gtm-vis-first-on-screen-50443292_563="523" data-gtm-vis-has-fired-50443292_562="1" data-gtm-vis-has-fired-50443292_563="1" data-gtm-vis-total-visible-time-50443292_562="10000" data-gtm-vis-total-visible-time-50443292_563="10000" data-title="Abstract" lang="en" style="background-color: white; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif;"><div class="c-article-section" id="Abs1-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Abs1" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; color: #222222; font-size: 1.5rem; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;"><br /></h2><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Abs1" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; color: #222222; font-size: 1.5rem; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Abstract</h2><div class="c-article-section__content" id="Abs1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">This study aimed to evaluate the effects of propofol and dexmedetomidine over different timescales on the IVF outcomes for transvaginal oocyte retrieval (TVOR). Twenty-four rats included in the study were divided into two main groups and three subgroups were subjected to the ovulation induction process. Group 1 was administered propofol (100 mg/kg i.v.) and group 2 were administered dexmedetomidine (25 µg/kg i.p.) The oviduct collection procedure was completed within 15 min for subgroup Pro15min, Dex15min (n = 4), within 16 to 30 min for subgroup Pro30min, Dex30min (n = 4) and within 31 to 60 min for subgroup Pro60min, Dex60min (n = 4) after euthanasia. The total number of oocytes was counted. After in vitro fertilization, the number and quality of embryos were evaluated. The number of pups born were evaluated after embryo transfer. The embryo number, quality and pup count decreased as the administration time for propofol increased (p < 0.05). No statistically significant difference was found between the dexmedetomidine subgroups for embryo number, quality and pup count(p > 0.05). As the exposure time to propofol increased, the number and quality of embryos obtained, and the pup count, decreased. The use of dexmedetomidine had no negative impacts on the number of embryos, their quality or the number of pups.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;"><a href="https://www.arvigen.com/2018/12/study-of-nature-and-origin-of-squirting.html" target="_blank"><span style="color: red; font-size: large;">Study of Nature and origin of "squirting" in female sexuality.</span></a><br /></p></div></div></section><section data-gtm-vis-first-on-screen-50443292_562="34910" data-gtm-vis-first-on-screen-50443292_563="34910" data-gtm-vis-total-visible-time-50443292_562="7500" data-gtm-vis-total-visible-time-50443292_563="7500" data-title="Introduction" style="background-color: white; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif;"><div class="c-article-section" id="Sec1-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec1" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; color: #222222; font-size: 1.5rem; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Introduction</h2><div class="c-article-section__content" id="Sec1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">In vitro fertilization (IVF) is currently the most widely used assisted reproductive technique worldwide. The IVF procedure technically includes ovarian stimulation and monitoring, oocyte collection with transvaginal follicle aspiration, fertilization in the laboratory and finally, transfer of embryos back to the uterus. Obtaining oocytes from ovarian follicles by ultrasound-guided needle aspiration from the vaginal wall requires anesthesia and/or analgesia, and this is defined as transvaginal oocyte retrieval (TVOR)<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 1" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-021-03177-z#ref-CR1" id="ref-link-section-d30305945e463" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Kwan, I. et al. Pain relief for women undergoing oocyte retrieval for assisted reproduction. Cochrane Database Syst Rev 5(5), CD004829 (2018).">1</a></span>. Although TVOR is a relatively simple and minimally invasive procedure, the patient may experience anxiety and pain due to puncture of the vaginal skin mucosa and ovarian capsule with a needle. Various methods of anesthesia are used to manage the patients, such as general anesthesia, conscious sedation and regional anesthesia. There is no consensus on the type of anesthetic agent to use for TVOR.</p><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Several studies have indicated that anesthetic drugs can enter the follicular fluids (FF)<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 4" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-021-03177-z#ref-CR4" id="ref-link-section-d30305945e477" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Christiaens, F. et al. Propofol concentrations in follicular fluid during general anaesthesia for transvaginal oocyte retrieval. Hum. Reprod. 14(2), 345–348 (1999).">4</a></span>. There is some concern about anesthetic drugs accumulating in FF and their negative impacts on fertilization rates and embryo development under general anesthesia.</p><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Propofol is a rapidly acting anesthetic agent with short induction and recovery times<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 5" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-021-03177-z#ref-CR5" id="ref-link-section-d30305945e484" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Boysen, K., et al., Comparison of induction with and first hour of recovery from brief propofol and methohexital anesthesia. 34(3): 212–215 (1990).">5</a></span>. The most commonly used anesthetic agent during TVOR is intravenous propofol, which is a safe drug for use in IVF.</p><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">However, some studies have revealed that propofol use is associated with reduced fertilization rates (FR) and inhibition of blastocyst development in single cell embryos<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 7" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-021-03177-z#ref-CR7" id="ref-link-section-d30305945e498" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Tatone, C. et al. An evaluation of propofol toxicity on mouse oocytes and preimplantation embryos. Hum. Reprod. 13(2), 430–435 (1998).">7</a></span>.</p><p style="box-sizing: inherit; color: #222222; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;"><span style="font-size: 18px;">Dexmedetomidine is a centrally acting α-2 receptor agonist with sedative and analgesic properties without respiratory suppressing effects. However, dexmedetomidine use may be limited by the increased incidence of hypotension and bradycardia, and a limited ability to achieve deep sedation</span><span style="font-size: 13.5px;">.</span><span style="font-size: 18px;"> There are not enough studies on the use of dexmedetomidine during the TVOR process.</span></p><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">This study aimed to evaluate the effect of these two anesthetic agents used in anesthesia for different durations on IVF results.</p><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;"><br /></p><div class="separator" style="clear: both; color: #222222; text-align: center;"><a href="https://blogger.googleusercontent.com/img/a/AVvXsEjV45GxezNDhUpWRH4Q1ld7Vy7-8GuJCl4yQaj8woyIOkncpge5z8uR2QSnC9fxZOm1KhJSNiPhXjDUcKRMOqBmacSVsh5nnen0Mq2k_xK8FRM3LtadgVvkbFp9DiPSCvtKp-kW8McbumBm43IKmI0eXRxnkbf1UlLQxvagnNSmFdF10zyY3Eyw671sMQ=s1600" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="800" data-original-width="1600" height="234" src="https://blogger.googleusercontent.com/img/a/AVvXsEjV45GxezNDhUpWRH4Q1ld7Vy7-8GuJCl4yQaj8woyIOkncpge5z8uR2QSnC9fxZOm1KhJSNiPhXjDUcKRMOqBmacSVsh5nnen0Mq2k_xK8FRM3LtadgVvkbFp9DiPSCvtKp-kW8McbumBm43IKmI0eXRxnkbf1UlLQxvagnNSmFdF10zyY3Eyw671sMQ=w645-h234" width="645" /></a></div><span style="color: red;"><br /></span><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;"><a href="https://www.arvigen.com/2021/06/natural-selection-increases-female.html"><span style="color: red; font-size: large;">Natural selection increases female fitness by reversing the exaggeration of a male sexually selected trait</span></a><br /></p></div></div></section><section data-gtm-vis-first-on-screen-50443292_562="38298" data-gtm-vis-first-on-screen-50443292_563="38298" data-gtm-vis-has-fired-50443292_562="1" data-gtm-vis-has-fired-50443292_563="1" data-gtm-vis-recent-on-screen-50443292_562="51261" data-gtm-vis-recent-on-screen-50443292_563="51261" data-gtm-vis-total-visible-time-50443292_562="10000" data-gtm-vis-total-visible-time-50443292_563="10000" data-title="Results" style="background-color: white; box-sizing: inherit;"><div class="c-article-section" id="Sec2-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec2" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1.5rem; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Results</h2><div class="c-article-section__content" id="Sec2-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">A total of 58 oocytes (50 metaphase II (MII), 4 metaphase I (MI) and 4 germinal vesicles (GV))—of which 44 fertilized—were collected in subgroup Pro15min. For subgroup Pro30min, 57 oocytes were obtained (49 MII, 5 MI, 3 GV), and 25 were fertilized. Fifty-six oocytes (49 MII, 3 MI, + GV) were obtained from subgroup Pro60min, and 11 of them were fertilized.</p><p style="box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">There were 31 s day embryos (SDE) in total (22 grade 1) from subgroup Pro15min, and 18 pups were obtained. For subgroup Pro30min, there were 14 s day embryos (SDE) in total (9 grade 1) and 6 pups were obtained. Eleven SDEs (2 grade 1) were from subgroup Pro60min, and 4 pups were obtained.</p><p style="box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">A total of 60 oocytes (51 MII, 5MI, 4GV) were collected in subgroup Dex15min, and 46 were fertilized. For subgroup Dex30min, 57 oocytes were obtained (51 MII, 2 MI, 4 GV) and were fertilized. Fifty-seven oocytes (47 MII, 6MI, 4 GV) were produced by subgroup Dex60min, and 11 of them were fertilized.</p><p style="box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">There were 34 SDEs (23 grade 1) from subgroup Dex15min, and 19 pups were obtained. For subgroup Dex30min, there was a total of 33 SDEs (25 grade 1) and 19 pups were obtained. Thirty-one SDEs (23 grade 1) were produced by subgroup Dex60min and 19 pups were obtained.</p><p style="box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">In our study, no statistically significant difference was found between the mean number of MII, MI, GV and total oocyte counts between the subgroups (P > 0.05) (Table 1).</p><header style="box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 18px;"><h1 class="c-article-table-title u-h1" id="table-1-title" style="box-sizing: inherit; font-size: min(max(1.5rem, 4vw), 2rem); letter-spacing: min(max(-0.01172rem, 4vw), -0.03906rem); line-height: 1.4; margin: 0px 0px 16px; padding: 0px; word-break: break-word;">Table 1 Comparison of oocyte maturation and fertilization numbers of propofol and dexmedetomidine groups.</h1><p class="c-article-table-subtitle" style="box-sizing: inherit; font-family: serif; font-size: 1.125rem; margin: 0px 0px 28px; padding: 0px;">From: The effect of Propofol versus Dexmedetomidine as anesthetic agents for oocyte pick-up on in vitro fertilization (IVF) outcomes</p></header><div class="c-article-table-container" style="box-sizing: inherit; margin: 0px 0px 24px; padding: 0px;"><div class="c-article-table-border c-table-scroll-wrapper" style="border: 1px solid rgb(213, 213, 213); box-sizing: inherit; color: #222222; font-family: serif; font-size: 18px; margin: 0px; padding: 0px; position: relative;"><div class="c-table-scroll-wrapper__content c-table-scroll-wrapper__fade--transparent" data-component-scroll-wrapper="" style="box-sizing: inherit; margin: 0px; min-width: 100%; overflow-x: auto; padding: 0px;"><table class="data last-table" style="border-collapse: collapse; border-spacing: 0px; border: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1rem; margin-bottom: 0px; width: 1246px;"><thead class="c-article-table-head" style="border-bottom: 5px solid rgb(149, 149, 149); box-sizing: inherit;"><tr style="border-bottom: 1px solid rgb(213, 213, 213); box-sizing: inherit;"><th class="u-text-left" style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Parameter</th><th class="u-text-left" style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Promin15 (n = 4)</th><th class="u-text-left" style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Promin30 (n = 4)</th><th class="u-text-left" style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Promin60 (n = 4)</th><th class="u-text-left" style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Dexmin15 (n = 4)</th><th class="u-text-left" style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Dexmin30 (n = 4)</th><th class="u-text-left" style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Dexmin60 (n = 4)</th><th class="u-text-left" style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">p value</th></tr></thead><tbody style="box-sizing: inherit;"><tr style="border-top: 0px; box-sizing: inherit;"><td class="u-text-left" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">M II</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">12.5 ± 0.58</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">12.25 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">12.25 ± .96</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">12.75 ± 1.26</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">12.75 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">11.75 ± 1.26</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.615</td></tr><tr style="border-top: 1px solid rgb(213, 213, 213); box-sizing: inherit;"><td class="u-text-left" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">M I-MII</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1 ± 0.82</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1.25 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.75 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1.25 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.5 ± 0.58</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1.5 ± 0.58</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.227</td></tr><tr style="border-top: 1px solid rgb(213, 213, 213); box-sizing: inherit;"><td class="u-text-left" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">GV</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1 ± 0.82</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.75 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1 ± 0.82</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1 ± 0 .82</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1 ± 0 .82</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1 ± 0.82</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.995</td></tr><tr style="border-top: 1px solid rgb(213, 213, 213); box-sizing: inherit;"><td class="u-text-left" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Total oocyte count</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">14.5 ± 0.58</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">14.25 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">14 ± 0</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">15.25 ± 0.96</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">14.25 ± 1.26</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">14.25 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.301</td></tr><tr style="border-top: 1px solid rgb(213, 213, 213); box-sizing: inherit;"><td class="u-text-left" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Fertilized GV</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.50 ± 0.58</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0 ± 0</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">00 ± 0</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.25 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.5 ± 0.58</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.5 ± 0.58</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.347</td></tr><tr style="border-top: 1px solid rgb(213, 213, 213); box-sizing: inherit;"><td class="u-text-left" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Fertilized MI-II</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1.750 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.750 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0 ± 0</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1.5 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1.25 ± 0 .5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1 ± 0.82</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.019</td></tr><tr style="border-top: 1px solid rgb(213, 213, 213); box-sizing: inherit;"><td class="u-text-left" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Fertilized MII</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">8.75 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">5.5 ± 0.57</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">2.75 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">9.75 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">9.25 ± 0.58</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">9.25 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.002*</td></tr><tr style="border-top: 1px solid rgb(213, 213, 213); box-sizing: inherit;"><td class="u-text-left" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Total number of fertilized oocytes</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">11 ± 0.82</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">6.25 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">2.75 ± 0.5</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">11.5 ± 1</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">11.25 ± 0..96</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">11.5 ± 0.58</td><td class="u-text-char" style="border-right: 1px solid rgb(213, 213, 213); box-sizing: inherit; line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.004*</td></tr></tbody></table></div></div><div class="c-article-table-footer" style="box-sizing: inherit; margin: 0px; padding: 10px;"><ol style="box-sizing: inherit; color: #222222; font-family: serif; font-size: 18px; list-style: none; margin: 0px; padding: 0px;"><li style="box-sizing: inherit;"><i style="box-sizing: inherit;">MII</i> metaphase II stage oocyte, <i style="box-sizing: inherit;">MI-II</i> matured from MI to MII oocyte, <i style="box-sizing: inherit;">GV</i> germinal vesicle, <i style="box-sizing: inherit;">Promin15</i> Propofol subgroup (0–15 min), <i style="box-sizing: inherit;">Promin30</i> Propofol subgroup (15–30 min), <i style="box-sizing: inherit;">Promin60</i> Propofol subgroup (30–60 min), <i style="box-sizing: inherit;">Dexmin15</i> Dexmedetomidine (0–15 min), <i style="box-sizing: inherit;">Dexmin30</i> Dexmedetomidine (15–30 min), <i style="box-sizing: inherit;">Dexmin60</i> Dexmedetomidine (30–60 min).</li><li style="box-sizing: inherit;">Groups were compared using the Kruskal Wallis test; paired comparisons were made using the Mann–Whitney U test in the parameters that differed (*P < 0.05).</li><li style="box-sizing: inherit;"><br /></li><li style="box-sizing: inherit;"><span face="-apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif">Fertilized MII, MI–MII oocyte counts and total fertilized oocyte counts for the subgroups are presented in Fig1.</span></li></ol><div style="color: #222222; font-size: 18px;"><span face="-apple-system, BlinkMacSystemFont, Segoe UI, Roboto, Oxygen-Sans, Ubuntu, Cantarell, Helvetica Neue, sans-serif"><br /></span></div><div><header style="box-sizing: inherit; color: #222222; font-size: 18px;"><h1 class="c-article-figure-title u-h1" data-test="top-caption" id="Fig1" style="box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: min(max(1.5rem, 4vw), 2rem); letter-spacing: min(max(-0.01172rem, 4vw), -0.03906rem); line-height: 1.4; margin: 0px 0px 16px; padding: 0px;">Figure 1</h1><p class="c-article-figure-subtitle" style="box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1.125rem; margin: 0px 0px 28px; padding: 0px;">From: The effect of Propofol versus Dexmedetomidine as anesthetic agents for oocyte pick-up on in vitro fertilization (IVF) outcomes</p></header><div class="c-article-figure-content" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><figure data-test="figure" style="box-sizing: inherit; margin: 0px;"><div class="c-article-figure-content-image" style="box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 18px; margin: 0px 0px 16px; padding: 0px; text-align: center;"><div class="c-article-figure-content-image-inline" style="box-sizing: inherit; display: inline-block; margin: 0px; padding: 0px;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/full/springer-static/image/art%3A10.1038%2Fs41598-021-03177-z/MediaObjects/41598_2021_3177_Fig1_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="Figure 1" aria-describedby="Fig1" height="741" src="https://media.springernature.com/full/springer-static/image/art%3A10.1038%2Fs41598-021-03177-z/MediaObjects/41598_2021_3177_Fig1_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="1419" /></picture></div></div><div class="c-article-figure-description" data-test="bottom-caption" id="figure-1-desc" style="box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; margin: 0px 0px 24px; padding: 0px;"><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px; padding: 0px;">Comparison of fertilized MII oocyte and total fertilized oocyte counts between groups. <i style="box-sizing: inherit;">Pro15min</i> Propofol subgroup (0–15 min), <i style="box-sizing: inherit;">Pro30min</i> Propofol subgroup (15–30 min), <i style="box-sizing: inherit;">Pro60min</i> Propofol subgroup (30–60 min), <i style="box-sizing: inherit;">Dex15min</i> Dexmedetomidine (0–15 min), <i style="box-sizing: inherit;">Dex30min</i> Dexmedetomidine (15–30 min), <i style="box-sizing: inherit;">Dex60min</i> Dexmedetomidine (30–60 min). There was no fertilization in MI oocytes in groups Pro30min and Pro60min. The groups were compared using the Kruskal Wallis test; paired comparisons were made using the Mann–Whitney U test for the different parameters. (**P < 0.05).</p><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px; padding: 0px;"><br /></p><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">When the propofol groups were compared, there was a significant difference in terms of fertilized MII oocytes between the Pro15min subgroup, and the Pro30min and Pro60min subgroups (P = 0.017 and P = 0.015, respectively). This significant difference was also found in terms of the total fertilized oocyte count between the Pro15min subgroup, and the Pro30min and Pro60min subgroups (P = 0.017 for both).When the Pro30min and Pro60min subgroups were compared for the number of fertilized MII, MI and total fertilized oocytes, it was found to be significantly higher in the Pro30min group (P = 0.017, P = 0.040 and P = 0.015, respectively).</p><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">There was no statistically significant difference between dexmedetomidine subgroups in terms of fertilized MII, MI oocyte counts and total fertilized oocyte counts (P > 0.05) (Fig. 1).When the same time periods were compared, there was no statistically significant difference between the total fertilized oocyte count and fertilized MI oocytes between the Pro15min and Dex15min subgroups (P = 0.350 and P = 0.495, respectively); however, although the mean numbers of fertilized MII oocytes were close to each other, a statistically significant difference was found in favor of the Dex15min group (P = 0.040).When the Pro30min and Dex30min subgroups were compared in terms of fertilized MII, MI and total fertilized oocyte counts, statistically significant differences were found in favor of the Dex30min subgroup (P = 0.017 and P = 0.018, respectively).When the Pro60min and Dex60min subgroups were compared for the numbers of fertilized MII, MI and total fertilized oocytes, they were found to be statistically higher in the Dex60min subgroup (P = 0.015, P = 0.046 and P = 0.017, respectively).</p><p style="box-sizing: inherit; color: #222222; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">In this study, it was observed that the number of SDEs (Fig. 3), grade 1 embryos and the pups in the Pro15min subgroup were found to be statistically higher than the Pro30min and Pro60min subgroups (Fig. 2).</p><header style="box-sizing: inherit; color: #222222; font-size: 18px;"><h1 class="c-article-figure-title u-h1" data-test="top-caption" id="Fig2" style="box-sizing: inherit; font-size: min(max(1.5rem, 4vw), 2rem); letter-spacing: min(max(-0.01172rem, 4vw), -0.03906rem); line-height: 1.4; margin: 0px 0px 16px; padding: 0px;">Figure 2</h1><p class="c-article-figure-subtitle" style="box-sizing: inherit; font-size: 1.125rem; margin: 0px 0px 28px; padding: 0px;">From: The effect of Propofol versus Dexmedetomidine as anesthetic agents for oocyte pick-up on in vitro fertilization (IVF) outcomes</p></header><div class="c-article-figure-content" style="box-sizing: inherit; clear: both; color: #222222; font-size: 18px; margin: 0px; padding: 0px;"><figure data-test="figure" style="box-sizing: inherit; margin: 0px;"><div class="c-article-figure-content-image" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px; text-align: center;"><div class="c-article-figure-content-image-inline" style="box-sizing: inherit; display: inline-block; margin: 0px; padding: 0px;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/full/springer-static/image/art%3A10.1038%2Fs41598-021-03177-z/MediaObjects/41598_2021_3177_Fig2_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="Figure 2" aria-describedby="Fig2" height="782" src="https://media.springernature.com/full/springer-static/image/art%3A10.1038%2Fs41598-021-03177-z/MediaObjects/41598_2021_3177_Fig2_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="1419" /></picture></div></div><div class="c-article-figure-description" data-test="bottom-caption" id="figure-2-desc" style="box-sizing: inherit; margin: 0px 0px 24px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; padding: 0px;">Comparison of the number of second day embryos between groups, the number of grade 1 embryos, and the number of female and male offspring born. <i style="box-sizing: inherit;">Pro15min</i> Propofol subgroup (0–15 min), <i style="box-sizing: inherit;">Pro30min</i> Propofol subgroup (15–30 min), <i style="box-sizing: inherit;">Pro60min</i> Propofol subgroup (30–60 min), <i style="box-sizing: inherit;">Dex15min</i> Dexmedetomidine (0–15 min), <i style="box-sizing: inherit;">Dex30min</i> Dexmedetomidine (15–30 min), <i style="box-sizing: inherit;">Dex60min</i> Dexmedetomidine (30–60 min). There was no fertilization in MI oocytes in groups Pro15min and Pro30min. The groups were compared using the Kruskal Wallis test; paired comparisons were made using the Mann Whitney U test for the different parameters (* P < 0.05).</p><div><br /></div><div>When SDEs, grade 1 embryos and pup numbers were compared between the Pro15min and Dex15min subgroups, there was no difference, but significant differences were observed between the Pro30min and Dex30min subgroups (P = 0.019, P = 0.015 and P = 0.017, respectively). A similar difference was found when the Pro60min and Dex60min subgroups were compared (P = 0.015, P = 0.017 and P = 0.011, respectively) (Fig. 3).</div><div><br /></div><div><header style="box-sizing: inherit;"><h1 class="c-article-figure-title u-h1" data-test="top-caption" id="Fig3" style="box-sizing: inherit; font-size: min(max(1.5rem, 4vw), 2rem); letter-spacing: min(max(-0.01172rem, 4vw), -0.03906rem); line-height: 1.4; margin: 0px 0px 16px; padding: 0px;">Figure 3</h1><p class="c-article-figure-subtitle" style="box-sizing: inherit; font-size: 1.125rem; margin: 0px 0px 28px; padding: 0px;">From: The effect of Propofol versus Dexmedetomidine as anesthetic agents for oocyte pick-up on in vitro fertilization (IVF) outcomes</p></header><div class="c-article-figure-content" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><figure data-test="figure" style="box-sizing: inherit; margin: 0px;"><div class="c-article-figure-content-image" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px; text-align: center;"><div class="c-article-figure-content-image-inline" style="box-sizing: inherit; display: inline-block; margin: 0px; padding: 0px;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/full/springer-static/image/art%3A10.1038%2Fs41598-021-03177-z/MediaObjects/41598_2021_3177_Fig3_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="Figure 3" aria-describedby="Fig3" height="499" src="https://media.springernature.com/full/springer-static/image/art%3A10.1038%2Fs41598-021-03177-z/MediaObjects/41598_2021_3177_Fig3_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="1770" /></picture></div></div><div class="c-article-figure-description" data-test="bottom-caption" id="figure-3-desc" style="box-sizing: inherit; margin: 0px 0px 24px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; padding: 0px;">Images of the second day embryos belonging to the groups. (<span style="box-sizing: inherit; font-weight: bolder;">A</span>) Propofol subgroup 3 (Pro60min); only one out of four oocytes is fertilized; fragmentation is observed in the fertilized embryo (Grade 2 quality). The red arrowhead shows fragmentation. (<span style="box-sizing: inherit; font-weight: bolder;">B</span>) Propofol subgroup 1 (Pro15min) blastomeres of equal size and good quality embryos without fragmentation (Grade1 quality). (<span style="box-sizing: inherit; font-weight: bolder;">C</span>) Dexmedetomidine subgroup 1 (Dex15min), high division rate and high-quality embryos without fragmentation are seen (Grade 1 embryo).</p><p style="box-sizing: inherit; margin: 0px; padding: 0px;"><br /></p><p style="box-sizing: inherit; margin: 0px; padding: 0px;"><br /></p><p style="box-sizing: inherit; margin: 0px; padding: 0px;"><br /></p><section data-gtm-vis-first-on-screen-50443292_562="5297" data-gtm-vis-first-on-screen-50443292_563="5297" data-gtm-vis-has-fired-50443292_562="1" data-gtm-vis-has-fired-50443292_563="1" data-gtm-vis-total-visible-time-50443292_562="10000" data-gtm-vis-total-visible-time-50443292_563="10000" data-title="Discussion" style="box-sizing: inherit;"><div class="c-article-section" id="Sec3-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec3" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Discussion</h2><div class="c-article-section__content" id="Sec3-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">This study was conducted to evaluate the sensitivity of rat oocytes and embryos to anesthetics in order to observe the biological response for clinical studies on the effect of using propofol and dexmedetomidine as anesthetic agents on assisted reproductive technology results. In terms of providing effective conscious sedation and analgesia, Kwan et al. concluded that evidence from 21 randomized controlled trials did not support one particular method or technique over another during and after oocyte recovery<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">1</span>.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Due to the structure of sperm in humans, it is possible to obtain successful results with the ICSI procedure, especially if expert embryologists perform the procedure. However, the acrosome sperm structure is not compatible with the ICSI technique, as stated in rats and mice studies reported in published literature. Therefore, the IVF technique was preferred over ICSI in this study. In addition, there are studies in the literature showing that there is no difference between fertilization rates, embryo quality and pregnancy rates in studies on IVF and ICSI. In this current study, MII oocytes were primarily preferred for the IVF procedure. Studies have previously reported that incubated oocyte maturation completed in the MI phase produces a certain percentage of good quality embryos after fertilization. In this current study, the incubation and maturation of oocytes were completed in the MI phase. The study included those that completed their maturation and turned into MII oocytes for IVF. Likewise, GV oocytes were incubated for maturation. However, we did not include the rare fertilized oocytes formed after this incubation. The successful results in our fertilization groups, especially in the Dex subgroups, were similar to the IVF-ET results in published literature.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">With modern IVF technologies and with an expert operator, a human pick up lasts a few minutes. However, the oocyte retrieval process takes longer in patients who develop too many follicles—such as polycystic ovarian patients—compared to other patients. As body mass index increases, the mean time spent in the operating room significantly increases. To the best of our knowledge, this is the first study comparing the effect of dexmedetomidine and propofol on IVF results.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Previously published data have suggested a dose-dependent and time-dependent toxic effect that propofol has on the fertilization rates of oocytes. These authors concluded that a propofol-based anesthetic technique resulted in significant concentrations of this agent in the follicular fluid that was related to the dose of propofol administered and the duration of its use. Exposure of unfertilized oocytes to propofol results in a high degree of parthenogenetic activation.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">These observations point to a possible effect of anesthetic drugs on oocyte physiology and embryo development, and the quality and pregnancy outcomes after embryo transfer, which have not been sufficiently investigated so far. In this present study, the duration of anesthesia was negatively correlated with fertilization rates in the propofol group. It was observed that the negative effects of propofol on fertilization rate (FR), the number and quality of embryos, and the number of offspring obtained after embryo transfer, increased in direct proportion to the time spent under anesthesia (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-021-03177-z#Tab1" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;">1</a>). The fact that this situation was not observed in terms of the number of oocytes obtained suggests that this negative effect of propofol is due to the exposure during the oocyte recruitment. We found a higher dependency between longer durations of anesthesia (> 30 min) and embryo quality, FR and the number of offspring obtained after embryo transfer (Figs. 1, 2). As the time spent under anesthesia with propofol increased, the negative effects on fertilization, embryo development and the number of offspring obtained after embryo transfer increased even more. In contrast to the present study, time-dependent toxic effects of propofol on fertilization rates have been reported in mice<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 7" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-021-03177-z#ref-CR7" id="ref-link-section-d30305945e1220" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Tatone, C. et al. An evaluation of propofol toxicity on mouse oocytes and preimplantation embryos. Hum. Reprod. 13(2), 430–435 (1998).">7</a></span>. Tola<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 21" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-021-03177-z#ref-CR21" id="ref-link-section-d30305945e1224" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Tola, E. N. The effect of anesthetic agents for oocyte pick-up on in vitro fertilization outcome: A retrospective study in a tertiary center. Taiwan. J. Obstet. Gynecol. 58(5), 673–679 (2019).">21</a></span> concluded that the duration of anesthesia should be kept less than 30 min due to an association between the duration of the anesthesia and the lower implantation and clinical pregnancy rates. In this study, when the subgroups were compared on the basis of 0–15 min of administration for both agents, no difference was observed. However, the difference between the results of both agents was very prominent for the subgroups where 16–30 min and 31–60 min of exposure were evaluated. This result supports the idea that the shorter the exposure time, the less the expected side effects are. This suggests that propofol administration for anesthesia should be applied for as short a time as possible.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">However, there are studies showing no such side effects for propofol. Ben-Shlomo et al. concluded the time spent under anesthesia, as well as the total dose of propofol administered, are not associated with fertilization and embryo quality.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Elnabtity et al. concluded that dexmedetomidine, used as a part of conscious sedation for oocyte retrieval, offered a favorable analgesic technique during and after the procedure with significantly less requirements than removing propofol intraoperatively. It was observed that dexmedetomidine protects the ovarian tissue of the rat from ischemia and reperfusion (I/R) injury. Another study showed that dexmedetomidine has a protective effect on ovarian tissue against oxidative stress caused by pneumoperitoneum It is hypothesized that this protective effect of dexmedetomidine is mediated by the α-2 adrenergic receptors. Gu et al. showed that dexmedetomidine was able to decrease lung injury caused indirectly by kidney I/R via α2-adrenoceptor-related and independent mechanisms. This current study observed that the dexmedetomidine group was better than the propofol group in terms of the number and quality of embryos obtained. When dexmedetomidine subgroups were evaluated, there were none of the adverse effects observed in propofol subgroups with increased time spent under anesthesia and on FR, embryo number and quality, and the number of offspring obtained after embryo transfer. When evaluated in terms of the number of offspring obtained after embryo transfer, the dexmedetomidine group was superior to the propofol group (Fig. 2).</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">In this present study, no statistical differences were observed between embryo numbers and their quality, FR and the number of offspring obtained after embryo transfer of the dexmedetomidine subgroups (Fig. 2). This result showed that the dexmedetomidine exposure time does not affect the embryo number, quality and the number of offspring obtained after embryo transfer. This is thought to be the result of the protective effect of the dexmedetomidine and α-2 adrenergic receptors on ovarian tissue shown previously.</p></div></div></section><section data-gtm-vis-first-on-screen-50443292_562="22219" data-gtm-vis-first-on-screen-50443292_563="22219" data-gtm-vis-total-visible-time-50443292_562="7500" data-gtm-vis-total-visible-time-50443292_563="7500" data-title="Conclusions" style="box-sizing: inherit;"><div class="c-article-section" id="Sec4-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec4" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Conclusions</h2><div class="c-article-section__content" id="Sec4-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">As the exposure time to propofol used as an anesthetic agent for TVOR procedure increases, the number of embryos, their quality or the number of pups obtained after transfer decreases. This situation was not observed with the use of dexmedetomidine. The dexmedetomidine use had no adverse effect on the number of embryos, their quality or the number of pups obtained after transfer. When the results of this study were evaluated, it was thought that if propofol is used in TVOR applications, the procedure should be completed as soon as possible to reduce the exposure time to avoid the negative effects that may occur.</p></div></div></section><section data-gtm-vis-first-on-screen-50443292_562="24621" data-gtm-vis-first-on-screen-50443292_563="24621" data-gtm-vis-has-fired-50443292_562="1" data-gtm-vis-has-fired-50443292_563="1" data-gtm-vis-total-visible-time-50443292_562="10000" data-gtm-vis-total-visible-time-50443292_563="10000" data-title="Materials and methods" style="box-sizing: inherit;"><div class="c-article-section" id="Sec5-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec5" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Materials and methods</h2><div class="c-article-section__content" id="Sec5-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The study was conducted in Sakarya University’s SÜDETAM laboratory under the authority and approval of Sakarya University’s experimental animal ethics committee on 06/01/2021 under decision No: 06. All experiments were performed in accordance with ARRIVE guidelines, and all procedures were performed in accordance with the relevant guidelines. Twenty-four adult female Sprague Dawley rats (weight 200–250 g; age 65–75 days) were provided by the Sakarya University Animal Reproduction Center and housed in groups with ad libitum food and water in the Animal Laboratory of Sakarya University. The holding room was maintained at a room temperature of 22 ± 2 °C with humid conditions (45–55%) and a 12 h light/day cycle.</p><h3 class="c-article__sub-heading" id="Sec6" style="box-sizing: inherit; font-size: 1.5rem; font-weight: 400; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Study design</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The stage of the rats’ estrous cycle was determined by performing daily vaginal smears after acclimation. Rats determined to have at least three consecutive 4-day estrous cycles were prepared for in vitro fertilization (IVF). All the rats were subjected to the IVF protocol to create hyperstimulation.</p><h3 class="c-article__sub-heading" id="Sec7" style="box-sizing: inherit; font-size: 1.5rem; font-weight: 400; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Stimulation and collection of oocytes</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Twenty-four rats were prepared according to the stimulus protocol. Both ovaries were stimulated through the intraperitoneal (i.p.) route in the female rats. For the first injection, using an intraperitoneal injection of 150–300 internal units (IU)/kg of pregnant mare serum gonadotropin (CHRONOGEST/PMSG, Intervet, Istanbul, Turkey) was followed approximately 48 h later by 150–300 IU/kg of human chorionic gonadotropin (hCG; Gonatropin, CHORULON, Intervet, Istanbul, Turkey). At 17–19 h after hCG administration, 15 IU of Pregnant Mare Serum Gonadotropin (PMSG) was administered.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The oocyte collection process was divided into four steps: Step 1, administration of the anesthetic agent used in the study; Step 2, waiting for the required time—according to the working group—followed by euthanasia (cervical dislocation) to extract the oviduct; Step 3, oocyte collection from the oviduct and Step 4, incubation of the oocytes until insemination.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The rats were then randomly divided into two different groups:</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Group 1 rats were intravenously administered propofol (Propofol 1% FRESENIUS, FRESENIUS Kabi, Istanbul, Turkey) at 100 mg/kg. Group 1 rats were randomly divided into three different subgroups and the oviduct collection procedure completed within 15 min for Group Pro15min (n = 4), within 16–30 min for Group Pro30min (n = 4) and within 31–60 min for Group Pro60min (n = 4).</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Group 2 rats were intraperitoneally administered a 25 µg/kg dexmedetomidine injection (PRECEDEX 200 µg/2 ml, Abbott, Istanbul, Turkey). Group 2 rats were also randomly divided into three different subgroups and the oviduct collection procedure completed within 0–15 min for Group Dex15min (n = 4), within 16–30 min for Group Dex30min (n = 4) and within 31–60 min for Group Dex60min (n = 4).</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">For oocyte pick-up, an aseptic technique was used to make a ventral midline incision to expose the reproductive organs and remove the oviducts. In this way the oocytes were collected from the removed ovaries. Oocytes were classified into the germinal vesicle (GV), the metaphase I (MI) and metaphase II (MII) stage. To compare meiotic progression during oocyte maturation in different systems, the average time for each stage of nuclear progression was calculated. This method was previously described by Sirard et al..</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">To incubate the oocytes, Human Tubal Fluid (HTF) medium (Cat. No. 90166, Irvine Scientific, USA) was cultured for one day before placing in an incubator at 37 °C in a 5% CO<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> environment. Sperm collection and preincubation processes for mature male rats (outbred rats aged 10 weeks to 10 months) before the oocyte collection were performed using established procedures previously described by Hino. The sperm were transferred to a droplet of HTF medium and incubated at 37 °C under 5% CO<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> in humidified air for preincubation.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">HTF medium (Cat. No. 90166, Irvine Scientific, USA) was used for sperm preincubation, fertilization and embryo transfer. Embryos were washed by passing through four such droplets which were kept at 37 °C under 5% CO<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span> in humidified air overnight.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The initial concentration of capacitated sperms was found to be approximately 1 × 10<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">6 ml</span>; the sperms were preincubated for approximately 15–60 min before being placed in fertilization drops and the final concentration of sperms was found to be approximately 4.5 to 6 × 10<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">5</span> million/ml. Then sperms were transferred to the fertilization drops with oocytes.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Cumulus-oocyte complexes were transferred to culture media containing sperm and incubated in culture medium for 10 h at 37 °C and 5% CO<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span>. After 10 h of incubation, the oocyte cumulus complexes were removed from the cumulus cells by taking the medium containing 0.1% Hyaluronidase. Oocytes with 2 pronuclei (2PN) and at least one sperm tail in the ooplasm were considered fertilized under an inverted microscope for fertilization control.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Approximately 10 h after the IVF procedure, the oocytes were washed with HTF medium, transferred to the culture medium, and taken into an embryo culture. Fertilization control at 10 h after IVF was performed under an inverted microscope to identify any polyspermic fertilization or parthenogenetic embryos. Fertilization control was performed by controlling male pronucleus (MPN) formation of oocytes at 200 or 400 magnifications. The ooplasm was checked for an entire and enlarged sperm head and MPN. Because it is challenging to distinguish MPN from female pronucleus, oocytes with three pronuclei, two pronuclei and a second polar body or without a second polar body were considered MPN. Oocytes with a female pronucleus and a second polar body, or oocytes with two female pronuclei without a second polar body were considered activated. At the 20th hour of embryo culture, 2-cell embryos were counted and embryos graded. These 2-cell embryos were also considered to be fertilized embryos.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Then, fertilization was checked and the fertilized oocytes were washed and transferred to culture drops, and the resulting embryos were monitored up to transfer<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 30" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-021-03177-z#ref-CR30" id="ref-link-section-d30305945e1369" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-thickness: 0.0625rem; text-underline-offset: 0.08em; vertical-align: baseline; word-break: break-word;" title="Agca, Y. & Critser, J. K. Chapter 7—Assisted Reproductive Technologies and Genetic Modifications in Rats. In The Laboratory Rat (Second Edition) (eds Suckow, M. A. et al.) 165–189 (Academic Press, 2006).">30</a></span>. The total number of cells was counted as an embryo evaluation criterion: the ratio of blastomeres size and fragmentation rates, as well as the number of blastomeres, were used as described by Ahumada.</p><h3 class="c-article__sub-heading" id="Sec8" style="box-sizing: inherit; font-size: 1.5rem; font-weight: 400; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Cryopreservation and thawing of embryos</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">After the collection of oocytes, female rats were sacrificed without being used for embryo transfer. Female rats from the same source and at the same age as the sacrificed female rats were used for transfer. Cryopreservation and thawing of second day embryos were performed using established procedures previously described by Nakao. Embryos were stored frozen for 1–3 weeks. A survival rate of 94% was found for cryopreserved embryos after thawing.</p><h3 class="c-article__sub-heading" id="Sec9" style="box-sizing: inherit; font-size: 1.5rem; font-weight: 400; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Transfer of embryos into oviducts</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Embryo transfer was performed using previously described procedures by Nakagata. Pseudo pregnant female rats were used that had been mated with vasoligated males. All pseudo pregnant female rats were anesthetized by an intramuscular administration of 50 mg/kg of ketamine hydrochloric acid (KETALAR; Eczacibasi Warner- Lambert Ilac Sanayi, Levent, Istanbul, Turkey) and 7 mg/kg of xylazine hydrochloric acid (ROMPUN, Bayer Sisli, Istanbul, Turkey). An aseptic technique was used to make a ventral midline incision to expose the reproductive organs and the oviducts. Then, the ovaries and oviducts were exposed and fat tissue near the ovaries was clamped for observation. Next, the embryos (sandwiched by air bubbles) were drawn into a capillary. The rats gave birth 20–21 days after surgery, and the numbers of pups were counted.</p><h3 class="c-article__sub-heading" id="Sec10" style="box-sizing: inherit; font-size: 1.5rem; font-weight: 400; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Statistics</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Statistical analyses were performed using SPSS 24.0 statistical software (SPSS Inc. and Lead Tech. Inc. Chicago. USA). The Kolmogorov–Smirnov test was used for normally distributed data. The Kruskal Wallis test was used for numerical data of the subgroups that did not show a normal distribution. Intergroup evaluations for statistically different parameters were performed using the Mann–Whitney <i style="box-sizing: inherit;">U</i> test and comparing them in pairs. Results are given as mean ± standard deviation. For all statistical analyses, a two-tailed p value < 0.05 was considered statistically significant.</p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/a/AVvXsEg50UHYNGtMF6iWygcCpV_JRrMoaIyL7F2EdrL5W-uhtMMRZ13hlWhka3mPavlbzy46go9G9GO3lizxiKkiKN3zv1N-L2rTCu9I2mBVA6eLC9srDJgjXZmGrN_jWHBusD9Z15x_0I-XElpUGo4Ce5ZSykk1xJjNZdWSscwJWMgmGKZvHv34Q3pVJwTvIg=s1117" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="1117" data-original-width="1003" height="320" src="https://blogger.googleusercontent.com/img/a/AVvXsEg50UHYNGtMF6iWygcCpV_JRrMoaIyL7F2EdrL5W-uhtMMRZ13hlWhka3mPavlbzy46go9G9GO3lizxiKkiKN3zv1N-L2rTCu9I2mBVA6eLC9srDJgjXZmGrN_jWHBusD9Z15x_0I-XElpUGo4Ce5ZSykk1xJjNZdWSscwJWMgmGKZvHv34Q3pVJwTvIg=w453-h320" width="453" /></a></div><br /><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;"><br /></p></div></div></section><div id="MagazineFulltextArticleBodySuffix" style="box-sizing: inherit; margin: 0px; padding: 0px;"><section aria-labelledby="Bib1" data-gtm-vis-first-on-screen-50443292_563="46656" data-gtm-vis-polling-id-50443292_563="2620" data-gtm-vis-recent-on-screen-50443292_563="46656" data-gtm-vis-total-visible-time-50443292_563="6900" data-title="References" style="box-sizing: inherit;"><div class="c-article-section" id="Bib1-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Bib1" style="border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 1.5rem; letter-spacing: -0.01172rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">References</h2><div class="c-article-section__content" id="Bib1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><div data-container-section="references" style="box-sizing: inherit; margin: 0px; padding: 0px;"><ol class="c-article-references" style="box-sizing: inherit; 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Biom</i></p></li></ol></div></div></div></section></div></div></figure></div></div></div></figure></div><div><span style="color: red;"><a href="https://www.arvigen.com/2021/06/covid-vaccines-and-breastfeeding-what.html" target="_blank"><span style="font-size: large;">COVID vaccines and breastfeeding: what the data say</span></a><br /></span></div><div><span style="color: red;"><br /></span></div><div><a href="https://www.arvigen.com/2018/12/biomarker-risk-score-may-identify-women.html" target="_blank"><span style="color: red; font-size: large;">Biomarker Risk Score May Identify Women at Risk for Gestational Diabetes</span></a><br /></div></div></figure></div></div></div></div></div></div></section>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com5tag:blogger.com,1999:blog-204734490966806252.post-30115669544876105422021-08-27T01:18:00.000-07:002021-08-27T01:18:01.095-07:00Corona Vaccine : Decrease in antibodies within few month after corona vaccination ? Study Indicates<span style="font-family: courier;"><b>Pfizer, AstraZeneca vaccine antibodies may reduce by 50% after 2-3 months: Lancet study</b></span><div><br /></div><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjX4QrcJJSe57OsO_ceJlIY8q6x6ESwhz2Q3npgg6fGiyLEx6398eW3XZYZfl6_nVGVlw6AUtOtnxd_i0HgGm0_tlKVSaY5uMyMiq4DiS6FR9JtbexzZzGpT_zrmIJi6khDolmsIehpB4XY/s807/images+%25286%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="380" data-original-width="807" height="151" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjX4QrcJJSe57OsO_ceJlIY8q6x6ESwhz2Q3npgg6fGiyLEx6398eW3XZYZfl6_nVGVlw6AUtOtnxd_i0HgGm0_tlKVSaY5uMyMiq4DiS6FR9JtbexzZzGpT_zrmIJi6khDolmsIehpB4XY/w351-h151/images+%25286%2529.jpeg" width="351" /></a></div><br /><div><br /></div><div><h2 style="border: none; box-sizing: border-box; color: #111111; font-size: 17px; font-style: italic; font-weight: 400; line-height: 24px; margin: 0px; outline: 0px; padding: 10px 0px 0px;"><span style="font-family: times;">Antibody levels after completing immunisation with Pfizer and AstraZeneca vaccines may be reduced by over 50 per cent after 2-3 months or 10 weeks, a Lancet study has found.</span></h2></div><div><span style="font-family: times;"><br /></span></div><div><span style="font-family: times;"><a href="https://www.arvigen.com/2021/06/covid-vaccines-and-breastfeeding-what.html?m=1">COVID vaccines and breastfeeding: what the data say</a><br /></span></div><div><br /></div><div><span style="font-family: times;"><i>A study published in The Lancet journal has found that the total antibody levels start to become weaker six weeks after completing immunisation with Pfizer and AstraZeneca vaccines. The antibody levels may also be reduced by more than 50 per cent over 10 weeks or 2-3 months.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div><div><span style="font-family: times;"><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh3Q8K_3gAGFYvb-pVCtdMMweCG7m_za0mFEYcu0DZdAjDuPJRGbn_wTvkBemeM5e8zSImZMUJHoTJ-aMaWiKvMx-DDHy-BaPTvQ3QI7qZ0NoHivSZuDLb2S9zpDVMcz4HfKeWW-q-A4lQ8/s685/images+%25287%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="448" data-original-width="685" height="209" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh3Q8K_3gAGFYvb-pVCtdMMweCG7m_za0mFEYcu0DZdAjDuPJRGbn_wTvkBemeM5e8zSImZMUJHoTJ-aMaWiKvMx-DDHy-BaPTvQ3QI7qZ0NoHivSZuDLb2S9zpDVMcz4HfKeWW-q-A4lQ8/s320/images+%25287%2529.jpeg" width="320" /></a></div><br /><i><br /></i></span></div><div><span style="font-family: times;"><i>According to researchers from University College London (UCL) in the UK, if the antibody levels carry on dropping at this rate, there are concerns that the protective effects of the vaccines may also begin to wear off, particularly against new variants. But how soon that might happen cannot be predicted yet, they added.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div><div><span style="font-family: times;"><i>The UCL Virus Watch study also found that antibody levels are substantially higher following two doses of the Pfizer vaccine compared to two shots of the AstraZeneca jab, known as Covishield in India.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div><div><span style="font-family: times;"><i>Antibody levels were also much higher in vaccinated people than those with prior SARS-CoV-2 infection, they said.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div><div><span style="font-family: times;"><i>"The levels of antibody following both doses of either the AstraZeneca or Pfizer vaccine were initially very high, which is likely to be an important part of why they are so protective against severe Covid-19," said Madhumita Shrotri from UCL Institute of Health Informatics in a statement.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div><div><span style="font-family: times;"><i>However, we found these levels dropped substantially over the course of two to three months," Madhumita Shotri added.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div><div><span style="font-family: times;"><i>Findings consistent across all groups of people</i></span></div><div><span style="font-family: times;"><i>Based on data from more than 600 people aged 18 and above, the findings of the study were consistent across all groups of people, regardless of age, chronic illnesses or sex, researchers said.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div><div><span style="font-family: times;"><i>The authors highlighted that although the clinical implications of waning antibody levels are not yet clear, some decline was expected and current research shows that vaccines remain effective against severe disease.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div><div><span style="font-family: times;"><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEggKfVCfqdXvncOogVQO8gHvFZ0mdn5EkXazPiaXRkiotYAQS1pE7dLx7K02xcpwdCh-24afV4Pl_WIaaT6LZoB6KnXHfTWMj7vEGkBGlbm9ma9XOJ9RhkeieEQOsxIJ_h4XjGgA2Fg70ZJ/s707/images+%25285%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="434" data-original-width="707" height="196" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEggKfVCfqdXvncOogVQO8gHvFZ0mdn5EkXazPiaXRkiotYAQS1pE7dLx7K02xcpwdCh-24afV4Pl_WIaaT6LZoB6KnXHfTWMj7vEGkBGlbm9ma9XOJ9RhkeieEQOsxIJ_h4XjGgA2Fg70ZJ/s320/images+%25285%2529.jpeg" width="320" /></a></div><br /><i><br /></i></span></div><div><span style="font-family: times;"><i>For Pfizer, antibody levels declined from a median of 7506 units per millilitre (U/mL) at 2141 days, to 3320 U/mL on 70 or more days.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div><div><span style="font-family: times;"><i>For the AstraZeneca vaccine, antibody levels were reduced from a median of 1201 U/mL at 020 days to 190 U/mL at 70 or more days, over a five-fold reduction.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div><div><span style="font-family: times;"><i>"When we are thinking about who should be prioritised for booster doses, our data suggests that those vaccinated earliest, particularly with the AstraZeneca vaccine, are likely to now have the lowest antibody levels," said Professor Rob Aldridge from UCL Institute of Health Informatics.</i></span></div><div><span style="font-family: times;"><i><br /></i></span></div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com2tag:blogger.com,1999:blog-204734490966806252.post-24500246239264180012021-08-12T22:33:00.004-07:002021-08-12T22:33:52.757-07:00Naturally enhanced neutralizing breadth against SARS-CoV-2 one year after infection<p> </p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg2BH5dHTX1bxK9fPvHg_pcz6q79wTvWq4uYxoFIuDsVGF10JrW59NROxM0nRszVo1FoXWTqG3eBfQq9l2udJgzwqTzXAbpvRcVAx_5JjYP3B8qk5Nn2sftuwqdGLINe-s6nud-fQ4FkWsH/s700/the-breath-is-a-powerful-tool-that-can-impact-our-health-and-well-being.-.jpg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="400" data-original-width="700" height="329" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg2BH5dHTX1bxK9fPvHg_pcz6q79wTvWq4uYxoFIuDsVGF10JrW59NROxM0nRszVo1FoXWTqG3eBfQq9l2udJgzwqTzXAbpvRcVAx_5JjYP3B8qk5Nn2sftuwqdGLINe-s6nud-fQ4FkWsH/w666-h329/the-breath-is-a-powerful-tool-that-can-impact-our-health-and-well-being.-.jpg" width="666" /></a></div><br /><div class="separator" style="clear: both; text-align: center;"><br /></div><br /><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 2.4rem; letter-spacing: -0.01875rem;">Abstract</span></p><section aria-labelledby="Abs1" data-gtm-vis-first-on-screen-10482319_393="433" data-gtm-vis-first-on-screen-10482319_401="433" data-gtm-vis-has-fired-10482319_393="1" data-gtm-vis-has-fired-10482319_401="1" data-gtm-vis-total-visible-time-10482319_393="10000" data-gtm-vis-total-visible-time-10482319_401="10000" data-title="Abstract" lang="en" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Abs1-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><div class="c-article-section__content" id="Abs1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">More than one year after its inception, the coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains difficult to control despite the availability of several working vaccines. Progress in controlling the pandemic is slowed by the emergence of variants that appear to be more transmissible and more resistant to antibodie. Here we report on a cohort of 63 individuals who have recovered from COVID-19 assessed at 1.3, 6.2 and 12 months after SARS-CoV-2 infection, 41% of whom also received mRNA vaccines. In the absence of vaccination, antibody reactivity to the receptor binding domain (RBD) of SARS-CoV-2, neutralizing activity and the number of RBD-specific memory B cells remain relatively stable between 6 and 12 months after infection. Vaccination increases all components of the humoral response and, as expected, results in serum neutralizing activities against variants of concern similar to or greater than the neutralizing activity against the original Wuhan Hu-1 strain achieved by vaccination of naive individuals. The mechanism underlying these broad-based responses involves ongoing antibody somatic mutation, memory B cell clonal turnover and development of monoclonal antibodies that are exceptionally resistant to SARS-CoV-2 RBD mutations, including those found in the variants of concern. In addition, B cell clones expressing broad and potent antibodies are selectively retained in the repertoire over time and expand markedly after vaccination. The data suggest that immunity in convalescent individuals will be very long lasting and that convalescent individuals who receive available mRNA vaccines will produce antibodies and memory B cells that should be protective against circulating SARS-CoV-2 variants.</p></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="7257" data-gtm-vis-first-on-screen-10482319_401="7257" data-gtm-vis-total-visible-time-10482319_393="4900" data-gtm-vis-total-visible-time-10482319_401="4900" data-title="Main" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Sec1-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec1" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;"><a href="https://www.arvigen.com/2021/07/mix-and-match-covid-vaccines-case-is.html">Mix-and-match COVID vaccines: the case is growing, but questions remain</a><br /></h2><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec1" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Main</h2><div class="c-article-section__content" id="Sec1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">We initially characterized immune responses to SARS-CoV-2 in a cohort of patients who have recovered from COVID-19 infection (hereafter referred to as convalescent individuals) 1.3 and 6.2 months after infection. Between 8 February and 26 March 2021, 63 participants between the ages of 26 and 73 years old (median 47 years old) returned for a 12-month follow-up visit. Among those, 26 (41%) had received at least one dose of either the Moderna (mRNA-1273) or Pfizer-BioNTech (BNT162b2) vaccines, on average 40 days (range 2–82 days) before their study visit and 311 days (range 272–373 days) after the onset of acute illness (Supplementary Table <a data-track-action="supplementary material anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#MOESM2" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">1</a>). Participants were almost evenly split between the sexes (43% female) and of the individuals who returned for the 12-month follow-up, only 10% had been hospitalized and the remainder had experienced relatively mild initial infections. Only 14% of the individuals reported persistent long-term symptoms after 12 months, reduced from 44% at the 6-month time point<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 4" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR4" id="ref-link-section-d21789e821" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Gaebler, C. et al. Evolution of antibody immunity to SARS-CoV-2. Nature 591, 639–644 (2021).">4</a></span>. Symptom persistence was not associated with the duration and severity of acute disease or with vaccination status (Extended Data Fig. 1a–c). All participants tested negative for active infection at the 12-month time point as measured by a saliva-based PCR assay<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 4" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR4" id="ref-link-section-d21789e828" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Gaebler, C. et al. Evolution of antibody immunity to SARS-CoV-2. Nature 591, 639–644 (2021).">4</a></span>. The demographics and clinical characteristics of the participants are shown in Supplementary Tables 1, 2.</p></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="8728" data-gtm-vis-first-on-screen-10482319_401="8728" data-gtm-vis-has-fired-10482319_393="1" data-gtm-vis-has-fired-10482319_401="1" data-gtm-vis-total-visible-time-10482319_393="10000" data-gtm-vis-total-visible-time-10482319_401="10000" data-title="Plasma SARS-CoV-2 antibody reactivity" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif;"><div class="c-article-section" id="Sec2-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec2" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Plasma SARS-CoV-2 antibody reactivity</h2><div class="c-article-section__content" id="Sec2-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Antibody reactivity in plasma to the RBD and nucleoprotein (N) were measured by enzyme-linked immunosorbent assay (ELISA). We limited our analysis to RBD because plasma RBD antibodies are strongly correlated with neutralizing activity. Convalescent participants who had not been vaccinated maintained most of their anti-RBD IgM (103%), IgG (82%) and IgA (72%) titres between 6 and 12 months after infection (Fig. 1a, Extended Data Fig. 2a–k). Consistent with previous reports, vaccination increased the plasma RBD antibody levels, with IgG titres increasing by nearly 30-fold compared with unvaccinated individuals (Fig. 1a, right). The two individuals who did not show an increase in antibody titre had been vaccinated only two days before sample collection. In contrast to anti-RBD antibody titres that were relatively stable, anti-N antibody titres decreased significantly between 6 and 12 months in this assay, independently of vaccination status (Fig. 1b, Extended Data Fig. 2l–n).</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-gtm-vis-first-on-screen-10482319_399="10005" data-gtm-vis-has-fired-10482319_399="1" data-gtm-vis-total-visible-time-10482319_399="10000" data-test="figure" data-title="Plasma ELISAs and neutralizing activity." id="figure-1" style="border: 5px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; font-size: 18px; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig1" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Fig. 1: Plasma ELISAs and neutralizing activity.</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; display: inline-block; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9/figures/1" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: inline-block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-line: none; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41586-021-03696-9/MediaObjects/41586_2021_3696_Fig1_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure1" aria-describedby="Fig1" height="456" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41586-021-03696-9/MediaObjects/41586_2021_3696_Fig1_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-1-desc" style="box-sizing: inherit; font-size: 1.6rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px;"><span style="box-sizing: inherit; font-weight: bolder;">a</span>–<span style="box-sizing: inherit; font-weight: bolder;">d</span>, Plasma IgG antibody binding to SARS-CoV-2 RBD (<span style="box-sizing: inherit; font-weight: bolder;">a</span>) and N protein (<span style="box-sizing: inherit; font-weight: bolder;">b</span>) shown as area under the curve (AUC; numbers in red are mean geometric AUC), and plasma neutralizing activity (NT<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span>) in unvaccinated (<span style="box-sizing: inherit; font-weight: bolder;">c</span>) and vaccinated (vac) (<span style="box-sizing: inherit; font-weight: bolder;">d</span>) individuals 12 months after SARS-CoV-2 infection (<i style="box-sizing: inherit;">n</i> = 63). <i style="box-sizing: inherit;">n</i> = 63 individuals, 37 convalescent unvaccinated (black) and 26 convalescent vaccinated (blue) individuals. <span style="box-sizing: inherit; font-weight: bolder;">a</span>, <span style="box-sizing: inherit; font-weight: bolder;">b</span>, Two-sided Kruskal–Wallis test with subsequent Dunn’s multiple comparisons. <span style="box-sizing: inherit; font-weight: bolder;">c</span>, <span style="box-sizing: inherit; font-weight: bolder;">d</span>, Lines connect longitudinal samples from the same individual. Two-sided Friedman test with subsequent Dunn’s multiple comparisons. Two individuals who received their first dose of vaccine 24–48 h before sample collection are represented in purple. <span style="box-sizing: inherit; font-weight: bolder;">e</span>, Plasma neutralizing activity against indicated SARS-CoV-2 variants of concern (<i style="box-sizing: inherit;">n</i> = 30, 15 convalescent and 15 convalescent vaccinated individuals). The B.1.526 variant used here contains the E484K substitution. Substitutions, deletions and insertions in S variants used here are described in Methods. Two-tailed Mann–Whitney test. Red numbers in <span style="box-sizing: inherit; font-weight: bolder;">c</span>–<span style="box-sizing: inherit; font-weight: bolder;">e</span> indicate the geometric mean NT<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span> at the indicated time point. All experiments were performed at least in duplicate.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><p style="box-sizing: inherit; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;"><br /></p><h1 style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px; text-align: left;"><a href="https://www.arvigen.com/2021/06/covid-vaccines-and-breastfeeding-what.html"><span style="font-size: large;">COVID vaccines and breastfeeding: what the data say</span></a></h1><p style="box-sizing: inherit; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Plasma neutralizing activity in 63 participants was measured using a human immunodeficiency virus 1 (HIV-1) pseudotyped with the SARS-CoV-2 spike (S) protein (Fig. 1c, d, Extended Data Fig. 2o). Twelve months after infection, the geometric mean half-maximal neutralizing titre (NT<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">50</span>) for the 37 individuals who had not been vaccinated was 75, which was not significantly different from the NT<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">50</span> for the same individuals at 6.2 months after infection (Fig. 1c). By contrast, the vaccinated individuals showed a geometric mean NT<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">50</span> of 3,684, which was nearly 50-fold higher than that of unvaccinated individuals and slightly higher than the 30-fold increase in anti-RBD IgG antibodies (Fig. 1a, c, d). Neutralizing activity was directly correlated with IgG anti-RBD (Extended Data Fig. 2p) but not with anti-N titres (Extended Data Fig. 2q, r). We conclude that neutralizing titres remain relatively unchanged between 6 and 12 months after SARS-CoV-2 infection, and that vaccination further boosts this activity by nearly 50-fold.</p><p style="box-sizing: inherit; font-size: 18px; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">To determine the neutralizing activity against circulating variants of concern or interest, we performed neutralization assays on HIV-1 virus pseudotyped with the S protein of the following SARS-CoV-2 variants of concern or interest: B.1.1.7 (Alpha), B.1.351 (Beta), B.1.526 (Iota) and P.1 (Gamma). Twelve months after infection, neutralizing activity against the variants was generally lower than against wild-type SARS-CoV-2 virus in the same assay, with the greatest loss of activity against B.1.351 (Fig. 1e). After vaccination the geometric mean NT<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">50</span> increased to 11,493, 48,341, 22,109 and 26,553 against B.1.351, B.1.1.7, B.1.526 and P.1, respectively. These titres are an order of magnitude higher than the neutralizing titres that have been reported against wild-type SARS-CoV-2 at the peak of the initial response in infected individuals and in naive individuals receiving both doses of mRNA vaccines (Fig. 1d). Similar results were also obtained using authentic SARS-CoV-2 WA1/2020 and B.1.351 (Extended Data Fig. 2s).</p></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="18663" data-gtm-vis-first-on-screen-10482319_401="18663" data-gtm-vis-total-visible-time-10482319_393="5700" data-gtm-vis-total-visible-time-10482319_401="5700" data-title="Memory B cells" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Sec3-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec3" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Memory B cells</h2><div class="c-article-section__content" id="Sec3-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The memory B cell compartment serves as an immune reservoir containing a diverse collection of antibodies. Although antibodies to the N-terminal domain and other parts of S can also be neutralizing, we limited our analysis to memory B cells that produce anti-RBD antibodies because they are the most numerous and potent. To count RBD-specific memory B cells, we performed flow cytometry using biotin-labelled RBD<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 3" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR3" id="ref-link-section-d21789e1049" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Robbiani, D. F. et al. Convergent antibody responses to SARS-CoV-2 in convalescent individuals. Nature 584, 437–442 (2020).">3</a></span> (Fig. 2a, Extended Data Fig. 3a, b). Without vaccination, the number of RBD-specific memory B cells present 12 months after infection was 1.35-fold lower than the earlier 6.2-month time point (<i style="box-sizing: inherit;">P</i> = 0.027, Fig. 2a). By contrast, and consistent with previous reports, individuals who recovered from COVID-19 and received mRNA vaccines showed an average increase of 8.6-fold in the number of circulating RBD-specific memory B cells (Fig. 2a). We also counted B cells expressing antibodies that bound to both wild-type and K417N/E484K/N501Y mutant RBDs using flow cytometry (Extended Data Fig. 3c). The number of B cells cross-reacting with variant RBD was directly proportional to and 1.6- to 3.2-fold lower than the number of B cells binding to wild-type RBD (Fig. 2a).</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-gtm-vis-first-on-screen-10482319_399="68225" data-gtm-vis-total-visible-time-10482319_399="2200" data-test="figure" data-title="SARS-CoV-2 RBD-specific B cell memory." id="figure-2" style="border: 5px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig2" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Fig. 2: SARS-CoV-2 RBD-specific B cell memory.</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; display: inline-block; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9/figures/2" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: inline-block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-line: none; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41586-021-03696-9/MediaObjects/41586_2021_3696_Fig2_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure2" aria-describedby="Fig2" height="196" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41586-021-03696-9/MediaObjects/41586_2021_3696_Fig2_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-2-desc" style="box-sizing: inherit; font-size: 1.6rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px;"><span style="box-sizing: inherit; font-weight: bolder;">a</span>, Number of antigen-binding memory B cells per 2 × 10<span style="box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">6</span> B cells (Extended Data Fig. 5b, c) obtained at 1.3, 6.2 and 12 months after infection from 40 randomly selected individuals (vaccinated, <i style="box-sizing: inherit;">n</i> = 20; non-vaccinated, <i style="box-sizing: inherit;">n</i> = 20). Each dot represents one individual. Red horizontal bars indicate geometric mean values. Two-sided Kruskal–Wallis test with subsequent Dunn’s multiple comparisons. WT, wild type. <span style="box-sizing: inherit; font-weight: bolder;">b</span>, The distribution of antibody sequences from 6 individuals 1.3 (top) or 6.2 (middle) or 12 (bottom) months after infection. The number in the inner circle indicates the number of sequences analysed for the individual whose identifier is denoted above the circle. Pie slice size is proportional to the number of clonally related sequences. The outer black arc indicates the frequency of clonally expanded sequences detected in each participant. Coloured slices indicate persisting clones (same IGV and IGJ genes, with highly similar CDR3 sequences) found at both time points in the same participant. Grey slices indicate clones unique to the time point. White indicates sequences isolated once, and white slices indicate singlets found at both time points. <span style="box-sizing: inherit; font-weight: bolder;">c</span>, Number of somatic nucleotide mutations (SHM) in the IGVH and IGVL genes (Supplementary Table 3) obtained after 1.3 or 6.2 or 12 months (1.3 month, <i style="box-sizing: inherit;">n</i> = 889; 6.2 month, <i style="box-sizing: inherit;">n</i> = 975; 12 month, <i style="box-sizing: inherit;">n</i> = 1,105 (unvaccinated, <i style="box-sizing: inherit;">n</i> = 417; vaccinated, <i style="box-sizing: inherit;">n</i> = 688)). Red horizontal bars indicate mean values. Two-sided Kruskal–Wallis test with subsequent Dunn’s multiple comparisons.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The memory B cell compartment accumulates mutations and undergoes clonal evolution over the initial six months after infection. To determine whether the memory compartment continues to evolve between 6 and 12 months after infection, we obtained 1,105 paired antibody heavy- and light-chain sequences from 10 individuals who were also assessed at the earlier time points, 6 of whom were vaccinated (Fig. 2b, Extended Data Fig. 3d, Supplementary Table 3). There were few significant differences among the expressed IGHV and IGLV genes between vaccinated and un-vaccinated groups, or between the 1.3-, 6-month and 1-year time points (Extended Data Fig. 4a–c). <i style="box-sizing: inherit;">IGHV3-30</i> and <i style="box-sizing: inherit;">IGHV3-53</i> remained over-represented independently of vaccination status (Extended Data Fig. 4a).</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">All individuals assayed at 12 months showed expansion of RBD-binding memory cell clones that expressed closely related IGHV and IGLV genes (Fig. 2b, Extended Data Fig. 3d, e). The relative fraction of cells belonging to these clones varied from 7% to 54% of the repertoire, with no significant difference between vaccinated and non-vaccinated groups. The overall clonal composition differed between 6 and 12 months after infection in all individuals, suggesting ongoing clonal evolution (Fig. 2b, Extended Data Fig. 3d). Among the 89 clones found 12 months after infection, 61% were not previously detected and 39% were present at one of the earlier time points (Fig. 2b, Extended Data Fig. 3d). In vaccinated individuals, the increase in size of the memory compartment was paralleled by an increase in the absolute number of B cells representing all persistent clones (Extended Data Fig. 5a). Thus, RBD-specific memory B cell clones were re-expanded upon vaccination in all six convalescent individuals examined (Fig. 2b, Extended Data Figs. 3d, 5a).</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Somatic hypermutation of antibody genes continued between 6 and 12 months after infection (Fig. 2c). Slightly higher levels of antibody-gene mutation were found in individuals who had not been vaccinated compared with vaccinated individuals, possibly owing to recruitment of newly formed memory cells into the expanded memory compartment of the vaccinated individuals (Fig. 2c, Extended Data Fig. 5b). There was no significant difference in numbers of mutations between conserved and newly arising clones at the 12-month time point in vaccinated individuals (Extended Data Fig. 5c). Moreover, phylogenetic analysis revealed that sequences found at 6 and 12 months after infection were intermingled and similarly distant from their unmutated common ancestors (Extended Data Fig. 6). We conclude that clonal re-expansion of memory cells in response to vaccination is not associated with additional accumulation of large numbers of somatic mutations as might be expected if the clones were re-entering and proliferating in germinal centres.</p></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="70672" data-gtm-vis-first-on-screen-10482319_401="70672" data-gtm-vis-total-visible-time-10482319_393="3700" data-gtm-vis-total-visible-time-10482319_401="3700" data-title="Neutralizing activity of monoclonal antibodies" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Sec4-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec4" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Neutralizing activity of monoclonal antibodies</h2><div class="c-article-section__content" id="Sec4-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">To determine whether the antibodies obtained from memory B cells 12 months after infection bind to RBD, we performed ELISAs (Fig. 3a). We tested 174 antibodies, including: (1) 53 that were randomly selected from those that appeared only once and only after 1 year; (2) 91 that appeared as expanded clones or singlets at more than one time point; and (3) 30 representatives of newly arising expanded clones (Supplementary Tables 4, 5). Among the 174 antibodies tested, 173 bound to RBD, indicating that the flow cytometry method used to identify B cells expressing anti-RBD antibodies was efficient (Supplementary Tables 4, 5). The geometric mean ELISA half-maximal concentration (EC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">50</span>) of the antibodies obtained 12 months after infection was 2.6 ng ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span>, which was significantly lower than after 6 months, independently of vaccination status and suggestive of an increase in affinity (Fig. 3a, Extended Data Fig. 7a, b, Supplementary Tables 4, 5). Consistent with this observation, there was an overall increase in the apparent avidity of plasma antibodies between 1.3 and 12 months (<i style="box-sizing: inherit;">P</i> < 0.0001) (Extended Data Fig. 7c).</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-gtm-vis-first-on-screen-10482319_399="72075" data-gtm-vis-total-visible-time-10482319_399="2200" data-test="figure" data-title="Anti-SARS-CoV-2 RBD monoclonal antibodies." id="figure-3" style="border: 5px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig3" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Fig. 3: Anti-SARS-CoV-2 RBD monoclonal antibodies.</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; display: inline-block; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9/figures/3" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: inline-block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-line: none; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41586-021-03696-9/MediaObjects/41586_2021_3696_Fig3_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure3" aria-describedby="Fig3" height="264" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41586-021-03696-9/MediaObjects/41586_2021_3696_Fig3_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-3-desc" style="box-sizing: inherit; font-size: 1.6rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px;"><span style="box-sizing: inherit; font-weight: bolder;">a</span>, EC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span> for SARS-CoV-2 RBD of antibodies isolated at 1.3 (<i style="box-sizing: inherit;">n</i> = 152) 6.2 (<i style="box-sizing: inherit;">n</i> = 153) and 12 (<i style="box-sizing: inherit;">n</i> = 174) months after infection<span style="box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 3" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR3" id="ref-link-section-d21789e1335" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Robbiani, D. F. et al. Convergent antibody responses to SARS-CoV-2 in convalescent individuals. Nature 584, 437–442 (2020).">3</a>,<a aria-label="Reference 4" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR4" id="ref-link-section-d21789e1338" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Gaebler, C. et al. Evolution of antibody immunity to SARS-CoV-2. Nature 591, 639–644 (2021).">4</a></span>, determined by ELISA. Two-sided Kruskal–Wallis test with subsequent Dunn’s multiple comparisons (1.3 months versus 6.2 months, <i style="box-sizing: inherit;">P</i> = 0.27; 1.3 months versus 12 months, <i style="box-sizing: inherit;">P</i> = 0.0075; 6.2 versus 12 months, <i style="box-sizing: inherit;">P</i> < 0.0001). <span style="box-sizing: inherit; font-weight: bolder;">b</span>, SARS-CoV-2-neutralizing activity of monoclonal antibodies measured using a SARS-CoV-2 pseudovirus neutralization assay. IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span> values for antibodies isolated at 1.3, 6.2 and 12 months after infection against wild-type SARS-CoV-2 (Wuhan-Hu-1 strain) are shown. Each dot represents one antibody. Pie charts illustrate the fraction of non-neutralizing (IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span> > 1,000 ng ml<span style="box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span>) antibodies (grey slices), inner circle shows the number of antibodies tested per group. Horizontal bars and red numbers indicate geometric mean values. Statistical significance was determined through the two-sided Kruskal–Wallis test with subsequent Dunn’s multiple comparisons.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">All 174 RBD binding antibodies obtained from the 12-month time point were tested for neutralizing activity in a SARS-CoV-2 pseudotype-neutralization assay. When compared with the earlier time points from the same individuals, the geometric mean half-maximal inhibitory concentration (IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">50</span>) improved from 171 ng ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span> (at 1.3 months) to 116 ng ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span> (at 6 months) to 79 ng ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span> (at 12 months), with no significant difference between vaccinated and non-vaccinated individuals (Fig. 3b, Extended Data Fig. 7d, Supplementary Table 4). This increased potency was most evident in the antibodies expressed by expanded clones of B cells that were conserved for the entire observation period independently of vaccination status (<i style="box-sizing: inherit;">P</i> = 0.014) (Fig. 3b, right, Extended Data Fig. 7e–h, Supplementary Table 5). The overall increase in neutralizing activity among conserved clones was owing to accumulation of clones expressing antibodies with potent neutralizing activity and simultaneous loss of clones expressing antibodies with no measurable activity (<i style="box-sizing: inherit;">P</i> = 0.028) (Fig. 3b, bottom right). Consistent with this observation, antibodies obtained from clonally expanded B cells 12 months after infection were more potent than antibodies obtained from unique B cells at the same time point (<i style="box-sizing: inherit;">P</i> = 0.029) (Fig. 3b).</p></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="74054" data-gtm-vis-first-on-screen-10482319_401="74063" data-gtm-vis-total-visible-time-10482319_393="3800" data-gtm-vis-total-visible-time-10482319_401="3800" data-title="Epitopes and breadth of neutralization" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Sec5-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec5" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;"><a href="https://www.arvigen.com/2021/06/natural-selection-increases-female.html">Natural selection increases female fitness by reversing the exaggeration of a male sexually selected trait</a><br /></h2><div><br /></div><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec5" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Epitopes and breadth of neutralization</h2><div class="c-article-section__content" id="Sec5-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">To determine whether the loss of non-neutralizing antibodies over time was due to preferential loss of antibodies targeting specific epitopes, we performed biolayer interferometry (BLI) experiments in which a preformed antibody–RBD complex was exposed to a second monoclonal antibody targeting one of three classes of structurally defined epitopes (schematic in Fig. 4a). We assayed 60 randomly selected antibodies with comparable neutralizing activity from the 1.3- and 12-month time points. The 60 antibodies were evenly distributed between the two time points and between neutralizers and non-neutralizers (Fig. 4). Antibody affinities for RBD were similar among neutralizers and non-neutralizers obtained at the same time point (Fig. 4b, Extended Data Fig. 8). When the two sets of unrelated antibodies obtained 1.3 and 12 months after infection were compared, they showed increasing affinity over time independent of their neutralizing activity (Fig. 4b, Extended Data Fig. 8). In competition experiments, 2 out of 30 non-neutralizing antibodies inhibited binding of the class 1 (C105), 2 (C121 and C144) or 3 (C135) antibodies tested; the remaining 28 non-neutralizing antibodies must therefore bind to epitopes that do not overlap with the epitopes of these classes of antibodies (Fig. 4c, Extended Data Fig. 9). By contrast, 28 out of 30 neutralizing antibodies blocked class 1 or 2 antibodies whose target epitopes are structural components of the RBD that interact with its cellular receptor, angiotensin-converting enzyme 2 (ACE2) (Fig. 4c, Extended Data Fig. 9). In addition, whereas 9 of the 15 neutralizing antibodies obtained after 1.3 months blocked both class 1 and 2 antibodies, only 1 of the 15 obtained after 12 months did so. In contrast to the earlier time point, 13 of 15 neutralizing antibodies obtained after 12 months interfered only with C121, a class 2 antibody (Fig. 4c, Extended Data Fig. 9). We conclude that neutralizing antibodies are retained and non-neutralizing antibodies targeting RBD surfaces that do not interact with ACE2 are removed from the repertoire over time.</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-gtm-vis-first-on-screen-10482319_399="74789" data-gtm-vis-total-visible-time-10482319_399="2100" data-test="figure" data-title="Epitope targeting and evolution of anti-SARS-CoV-2 RBD antibodies." id="figure-4" style="border: 5px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig4" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Fig. 4: Epitope targeting and evolution of anti-SARS-CoV-2 RBD antibodies.</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; display: inline-block; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9/figures/4" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: inline-block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-line: none; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41586-021-03696-9/MediaObjects/41586_2021_3696_Fig4_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure4" aria-describedby="Fig4" height="955" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41586-021-03696-9/MediaObjects/41586_2021_3696_Fig4_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-4-desc" style="box-sizing: inherit; font-size: 1.6rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px;"><span style="box-sizing: inherit; font-weight: bolder;">a</span>, Schematic of the BLI experiment (left) and IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span> values for randomly selected neutralizing and non-neutralizing antibodies (Ab1 and Ab2) isolated at 1.3 and 12 months after infection (<i style="box-sizing: inherit;">n</i> = 15 antibodies per group, <i style="box-sizing: inherit;">n</i> = 60 antibodies in total). Red horizontal bars indicate geometric mean. Two-sided Mann–Whitney test. <span style="box-sizing: inherit; font-weight: bolder;">b</span>, Dissociation constants (<i style="box-sizing: inherit;">K</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">D</span>) of the <i style="box-sizing: inherit;">n</i> = 30 neutralizing (green) and <i style="box-sizing: inherit;">n</i> = 30 non-neutralizing (red) antibodies shown in <span style="box-sizing: inherit; font-weight: bolder;">a</span>. Horizontal bars indicate geometric mean values. Two-sided Kruskal–Wallis test with subsequent Dunn’s multiple comparisons. BLI traces are shown in Extended Data Fig. 8. <span style="box-sizing: inherit; font-weight: bolder;">c</span>, Heat map of relative inhibition of binding of a monoclonal antibody (Ab2) to preformed complexes of RBD with another monoclonal antibody (Ab1) (grey, no binding; orange, intermediate binding; red, high binding). Data are normalized by subtraction of the autologous antibody control. BLI traces are shown in Extended Data Fig. 9. <span style="box-sizing: inherit; font-weight: bolder;">d</span>, Neutralization of the indicated mutant RBD proteins with antibodies shown in <span style="box-sizing: inherit; font-weight: bolder;">a</span>–<span style="box-sizing: inherit; font-weight: bolder;">c</span>. Pie charts illustrate the fraction of antibodies that are poorly or non-neutralizing (IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span> of 100–1,000 ng ml<span style="box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span>, red), intermediate neutralizing (IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span> of 10–100 ng ml<span style="box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span>, pink) and potently neutralizing (IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span> of 0–10 ng ml<span style="box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span>, white) for each mutant. The number in the inner circle shows the number of antibodies tested. <span style="box-sizing: inherit; font-weight: bolder;">e</span>, Graphs show affinities (<i style="box-sizing: inherit;">y</i>-axis) plotted against neutralization activity (<i style="box-sizing: inherit;">x</i>-axis) for 18 clonal antibody pairs isolated 1.3 (top) and 12 months (bottom) after infection (<i style="box-sizing: inherit;">n</i> = 36 antibodies). Spearman correlation test. <span style="box-sizing: inherit; font-weight: bolder;">f</span>, BLI affinity measurements for same <i style="box-sizing: inherit;">n</i> = 36 paired antibodies as in <span style="box-sizing: inherit; font-weight: bolder;">e</span>. Two-tailed Wilcoxon test. <span style="box-sizing: inherit; font-weight: bolder;">g</span>, IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span> values for <i style="box-sizing: inherit;">n</i> = 30 paired neutralizing antibodies isolated at indicated time points versus indicated mutant SARS-CoV-2 pseudoviruses. Antibodies are divided into groups I, II and III (left), on the basis of neutralizing activity: I, potent clonal pairs that do not improve over time; II, clonal pairs that show increased activity over time; and III, clonal pairs showing decreased neutralization activity after 12 months. Antibody class assignment based on initial (1.3 month after infection) sensitivity to mutation is indicated on the right. Red stars indicate antibodies that neutralize all tested RBD mutants. Colour gradient indicates IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; vertical-align: baseline;">50</span> values ranging from 0 (white) to 1,000 ng ml<span style="box-sizing: inherit; font-size: 12px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span> (red).</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /><svg aria-hidden="true" class="u-icon" focusable="false" height="16" role="img" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div></figure></div><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">To determine whether there was an increase in neutralization breadth over time, the neutralizing activity of the 60 antibodies was assayed against a panel of RBD mutants covering residues associated with circulating variants of concern: R346S, K417N, N440K, A475V, E484K and N501Y (Fig. 4d, Supplementary Table 6). Increased activity was evident against K417N, N440K, A475V, E484K and N501Y (Fig. 4d, Supplementary Table 6). These data indicate that evolution of the antibody repertoire results in acquisition of neutralization breadth over time.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The increase in breadth and overall potency of memory B cell antibodies could be owing to shifts in the repertoire, clonal evolution or both. To determine whether changes in specific clones are associated with increases in affinity and breadth, we measured the relative affinity and neutralizing breadth of matched pairs of antibodies expressed by expanded clones of B cells that were maintained in the repertoire over the entire observation period. SARS-CoV-2-neutralizing activity of the antibodies present at 1.3 or 12 months was not significantly correlated with affinity at either time point when each time point was considered independently (Fig. 4e). However, there was a significant increase in overall affinity over time, including in the 4 pairs of antibodies with no measurable neutralizing activity (Fig. 4f and Supplementary Table 7). Neutralizing breadth was assayed for 15 randomly selected pairs of antibodies targeting epitopes assigned to the 3 dominant classes of neutralizing antibodies. Seven of the selected antibodies showed equivalent or decreased activity against wild-type SARS-CoV-2 after 12 months (Fig. 4g, Supplementary Table 8). However, neutralizing breadth increased between 1.3 and 12 months for all 15 pairs, even when neutralizing activity against the wild-type was unchanged or decreased (Fig. 4g, Supplementary Table 8). Only 1 out of the 15 antibodies obtained after 1.3 months neutralized all the mutants tested (Fig. 4g). By contrast, 10 out of the 15 antibodies obtained from the same clones 12 months after infection neutralized all variants tested, with IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">50</span> values as low as 1 ng ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span> against the RBD triple mutant K417N/E484K/N501Y found in B.1.351 (Fig. 4g, Supplementary Table 8). Similar results were obtained with authentic WA1/2020 and B.1.351 (Extended Data Fig. 7i). In conclusion, continued clonal evolution of anti-SARS-CoV-2 antibodies over 12 months favours increasing potency and breadth, resulting in monoclonal antibodies with exceptional activity against a wide group of variants.</p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjqNyjLf334Lp0R3EiiBpXdGMvAVdnRguV1OpVLkv583BG7WlnIzAR0rugeFw1xg3NSQsUG03TXBZYfBqQxCsc7UwaZb-1X7xSiliBnTU2NkS1aW75IwB7B36q8TBfRCSkOn0S_FZL0jYGF/s2536/Prayer.jpg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="1240" data-original-width="2536" height="265" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjqNyjLf334Lp0R3EiiBpXdGMvAVdnRguV1OpVLkv583BG7WlnIzAR0rugeFw1xg3NSQsUG03TXBZYfBqQxCsc7UwaZb-1X7xSiliBnTU2NkS1aW75IwB7B36q8TBfRCSkOn0S_FZL0jYGF/w637-h265/Prayer.jpg" width="637" /></a></div><br /><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;"><br /></p></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="77151" data-gtm-vis-first-on-screen-10482319_401="77151" data-gtm-vis-total-visible-time-10482319_393="2400" data-gtm-vis-total-visible-time-10482319_401="2400" data-title="Discussion" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Sec6-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec6" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;"><a href="https://www.arvigen.com/2018/12/study-of-nature-and-origin-of-squirting.html">Study of Nature and origin of "squirting" in female sexuality.</a><br /></h2><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec6" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;"><br /></h2><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec6" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Discussion</h2><div class="c-article-section__content" id="Sec6-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">During immune responses, activated B cells interact with cognate T cells and begin dividing before selection into the plasma cell, memory or germinal centre B cell compartments, partly on the basis of their affinity for antigen. Whereas B cells expressing high-affinity antibodies are favoured to enter the long-lived plasma cell compartment, the memory compartment is more diverse and can develop directly from activated B cells or from a germinal centre. Memory cells emanating from a germinal centre carry more mutations than those that develop directly from activated B cells because they undergo additional cycles of division.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Consistent with the longevity of bone marrow plasma cells, infection with SARS-CoV-2 leads to persistent anti-RBD antibodies in serum, and corresponding neutralizing responses. Nearly 93% of the plasma neutralizing activity is retained between 6 and 12 months after infection. Vaccination boosts the neutralizing response by 1.5 orders of magnitude by inducing additional plasma cell differentiation from the memory B cell compartment. Recruitment of evolved memory B cells producing antibodies with broad and potent neutralizing activity into the plasma cell compartment is likely to account for the high serologic activity of vaccinated convalescent individuals against variants of concern.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Less is known about selection and maintenance of the memory B cell compartment. SARS-CoV-2 infection produces a memory compartment that continues to evolve more than 12 months after infection with accumulation of somatic mutations, emergence of new clones and increasing affinity, all of which are consistent with long-term persistence of germinal centres. The increase in activity against SARS-CoV-2 mutants parallels the increase in affinity and is consistent with the finding that increasing the apparent affinity of anti-SARS-CoV-2 antibodies by dimerization or by creating bi-specific antibodies also increases resistance to RBD mutations.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Continued antibody evolution in germinal centres requires antigen, which can be retained in these structures over long periods of time. In addition, SARS-CoV-2 protein and nucleic acids have been reported to remain in the gut for at least two months after infection. Regardless of the source of the antigen, antibody evolution favours epitopes overlapping with the ACE2-binding site on the RBD, possibly because these are epitopes that are preferentially exposed on trimeric S protein or virus particles.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Vaccination after SARS-CoV-2 infection increases the number of RBD-binding memory cells by more than an order of magnitude by recruiting new B cell clones into memory and expanding persistent clones. The persistent clones expand without accumulating large numbers of additional mutations, indicating that clonal expansion of human memory B cells does not require re-entry into germinal centres and occurs in the activated B cell compartment.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The notable evolution of neutralizing breadth after infection with SARS-CoV-2 and the robust enhancement of serologic responses and B cell memory achieved with mRNA vaccination suggests that convalescent individuals who are vaccinated should enjoy high levels of protection against emerging variants without a need to modify existing vaccines. If memory responses evolve in a similar manner in naive individuals who receive vaccines, additional appropriately timed boosting with available vaccines should lead to protective immunity against circulating variants.</p></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="78941" data-gtm-vis-first-on-screen-10482319_401="78941" data-gtm-vis-has-fired-10482319_393="1" data-gtm-vis-has-fired-10482319_401="1" data-gtm-vis-total-visible-time-10482319_393="10000" data-gtm-vis-total-visible-time-10482319_401="10000" data-title="Methods" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Sec7-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec7" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Methods</h2><div class="c-article-section__content" id="Sec7-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><h3 class="c-article__sub-heading" id="Sec8" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Data reporting</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">No statistical methods were used to predetermine sample size. The experiments were not randomized and the investigators were not blinded to allocation during experiments and outcome assessment.</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec9" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Study participants</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Previously enrolled study participants were asked to return for a 12-month follow-up visit at the Rockefeller University Hospital in New York between 8 February and 26 March 2021. Eligible participants were adults with a history of participation in both prior study visits of our longitudinal cohort study of COVID-19 recovered individuals. All participants had a confirmed history of SARS-CoV-2 infection, either diagnosed during the acute infection by PCR with reverse transcription (RT–PCR) or retrospectively confirmed by seroconversion. Exclusion criteria included presence of symptoms suggestive of active SARS-CoV-2 infection. Most study participants were residents of the Greater New York City tri-state region and were asked to return approximately 12 months after the time of onset of COVID-19 symptoms. Participants presented to the Rockefeller University Hospital for blood sample collection and were asked about potential symptom persistence since their 6.2-month study visit, laboratory-confirmed episodes of reinfection with SARS-CoV-2, and whether they had received any COVID-19-related treatment or SARS-CoV-2 vaccination in the interim. Study participants who had received COVID-19 vaccinations, were exclusively recipients of one of the two currently EUA-approved mRNA vaccines, Moderna (mRNA-1273) or Pfizer-BioNTech (BNT162b2), and individuals who received both doses did so according to current interval guidelines, namely 28 days (range 28–30 days) for Moderna and 21 days (range 21–23 days) for Pfizer-BioNtech. Detailed characteristics of the symptomology and severity of the acute infection, symptom kinetics, and the immediate convalescent phase (7 weeks post-symptom onset until 6.2-month visit) have previously been reported. Participants who presented with persistent symptoms attributable to COVID-19 were identified on the basis of chronic shortness of breath or fatigue, deficit in athletic ability and/or three or more additional long-term symptoms such as persistent unexplained fevers, chest pain, new-onset cardiac sequalae, arthralgias, impairment of concentration/mental acuity, impairment of sense of smell/taste, neuropathy or cutaneous findings as previously described. Clinical data collection and management were carried out using the software iRIS by iMedRIS. All participants at Rockefeller University provided written informed consent before participation in the study and the study was conducted in accordance with Good Clinical Practice. For detailed participant characteristics see Supplementary Table 2. The study was performed in compliance with all relevant ethical regulations and the protocol (DRO-1006) for studies with human participants was approved by the Institutional Review Board of the Rockefeller University.</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec10" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">SARS-CoV-2 molecular tests</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Saliva was collected into guanidine thiocyanate buffer as previously described. RNA was extracted using either a column-based (Qiagen QIAmp DSP Viral RNA Mini Kit, catalogue (cat.) no. 61904) or a magnetic bead-based method as previously described. Reverse-transcribed cDNA was amplified using primers and probes validated by the CDC or by Columbia University Personalized Medicine Genomics Laboratory, respectively and approved by the FDA under the Emergency Use Authorization. Viral RNA was considered detected if <i style="box-sizing: inherit;">C</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">t</span> for two viral primers/probes were <40.</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec11" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Blood samples processing and storage</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Peripheral blood mononuclear cells obtained from samples collected at Rockefeller University were purified as previously reported by gradient centrifugation and stored in liquid nitrogen in the presence of FCS and DMSO. Heparinized plasma and serum samples were aliquoted and stored at −20 °C or less. Prior to experiments, aliquots of plasma samples were heat-inactivated (56 °C for 1 h) and then stored at 4 °C.</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec12" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">ELISAs</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">ELISAs to evaluate antibodies binding to SARS-CoV-2 RBD and N were performed by coating of high-binding 96-half-well plates (Corning 3690) with 50 μl per well of a 1 μg ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span> protein solution in PBS overnight at 4 °C. Plates were washed 6 times with washing buffer (1× PBS with 0.05% Tween-20 (Sigma-Aldrich)) and incubated with 170 μl per well blocking buffer (1× PBS with 2% BSA and 0.05% Tween-20 (Sigma)) for 1 h at room temperature. Immediately after blocking, monoclonal antibodies or plasma samples were added in PBS and incubated for 1 h at room temperature. Plasma samples were assayed at a 1:66 starting dilution and 7 (IgA and IgM anti-RBD) or 11 (IgG anti-RBD) additional threefold serial dilutions. Monoclonal antibodies were tested at 10 μg ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span> starting concentration and 10 additional fourfold serial dilutions. Plates were washed 6 times with washing buffer and then incubated with anti-human IgG, IgM or IgA secondary antibody conjugated to horseradish peroxidase (HRP) (Jackson Immuno Research 109-036-088 109-035-129 and Sigma A0295) in blocking buffer at a 1:5,000 dilution (IgM and IgG) or 1:3,000 dilution (IgA). Plates were developed by addition of the HRP substrate, TMB (ThermoFisher) for 10 min (plasma samples) or 4 min (monoclonal antibodies). The developing reaction was stopped by adding 50 μl 1 M H<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2</span>SO<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> and absorbance was measured at 450 nm with an ELISA microplate reader (FluoStar Omega 5.11, BMG Labtech) with Omega MARS software for analysis. For plasma samples, a positive control (plasma from participant COV72, diluted 66.6-fold and seven additional threefold serial dilutions in PBS) was added to every assay plate for validation. The average of its signal was used for normalization of all of the other values on the same plate with Excel software before calculating the area under the curve using Prism v.9.1(GraphPad). For monoclonal antibodies, the EC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">50</span> was determined using four-parameter nonlinear regression (GraphPad Prism v.9.1).</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec13" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Proteins</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Mammalian expression vectors encoding the RBDs of SARS-CoV-2 (GenBank MN985325.1; S protein residues 319-539) or K417N, E484K, N501Y RBD mutants with an N-terminal human IL-2 or Mu phosphatase signal peptide were previously described. SARS-CoV-2 nucleocapsid protein (N) was purchased from Sino Biological (40588-V08B).</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec14" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">SARS-CoV-2 pseudotyped reporter virus</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">A panel of plasmids expressing RBD-mutant SARS-CoV-2 S proteins in the context of pSARS-CoV-2-S<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">Δ19</span> has previously been described. Variant pseudoviruses resembling variants of concern B.1.1.7 (first isolated in the UK), B.1.351 (first isolated in South Africa), B.1.526 (first isolated in New York City) and P.1 (first isolated in Brazil) were generated by introduction of substitutions using synthetic gene fragments (IDT) or overlap extension PCR mediated mutagenesis and Gibson assembly. Specifically, the variant-specific deletions and substitutions introduced were: B.1.1.7: ΔH69/V70, ΔY144, N501Y, A570D, D614G, P681H, T761I, S982A, D1118H; B.1.351: D80A, D215G, L242H, R246I, K417N, E484K, N501Y, D614G, A701V; B.1.526: L5F, T95I, D253G, E484K, D614G, A701V; P.1: L18F, R20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The E484K and K417N/E484K/N501Y (KEN) substitution, as well as the deletions and substitutions corresponding to variants of concern were incorporated into an S protein that also includes the R683G substitution, which disrupts the furin cleaveage site and increases particle infectivity. Neutralizing activity against mutant pseudoviruses were compared to a wild-type SARS-CoV-2 S sequence (NC_045512), carrying R683G where appropriate.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">SARS-CoV-2 pseudotyped particles were generated as previously described. In brief, 293T cells were transfected with pNL4-3ΔEnv-nanoluc and pSARS-CoV-2-S<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">Δ19</span>, particles were collected 48 h after transduction, filtered and stored at −80 °C.</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec15" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Microneutralization assay with authentic SARS-CoV-2</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Microneutralization assays of SARS-CoV-2 virus were performed as previously described<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 3" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR3" id="ref-link-section-d21789e1941" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Robbiani, D. F. et al. Convergent antibody responses to SARS-CoV-2 in convalescent individuals. Nature 584, 437–442 (2020).">3</a></span>. The day before infection, Vero E6 cells were seeded at 1 × 10<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">4</span> cells per well into 96-well plates. The diluted plasma and antibodies were mixed with SARS-CoV-2 WA1/2020 or the B.1.351 variant and incubated for 1 h at 37 °C. The antibody–virus mix was then directly applied to Vero E6 cells and incubated for 22 h at 37 °C. Cells were subsequently fixed by adding an equal volume of 70% formaldehyde to the wells, followed by permeabilization with 1% Triton X-100 for 10 min. After washing, cells were incubated for 1 h at 37 °C with blocking solution of 5% goat serum in PBS (catalogue no. 005–000-121; Jackson ImmunoResearch). A rabbit polyclonal anti-SARS-CoV-2 nucleocapsid antibody (catalogue no. GTX135357; GeneTex) was added to the cells at 1:1,000 dilution in blocking solution and incubated at 4 °C overnight. Goat anti-rabbit AlexaFluor 594 (catalogue no. A-11012; Life Technologies) was used as a secondary antibody at a dilution of 1:2,000. Nuclei were stained with Hoechst 33342 (catalogue no. 62249; Thermo Fisher Scientific) at 1 μg ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span>. Images were acquired with a fluorescence microscope and analysed using ImageXpress Micro XLS (Molecular Devices). All experiments were performed in a biosafety level 3 laboratory.</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec16" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Pseudotyped virus neutralization assay</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Fourfold serially diluted plasma from COVID-19-convalescent individuals or monoclonal antibodies were incubated with SARS-CoV-2 pseudotyped virus for 1 h at 37 °C. The mixture was subsequently incubated with 293T<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">Ace2</span> cells<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 3" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR3" id="ref-link-section-d21789e1959" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Robbiani, D. F. et al. Convergent antibody responses to SARS-CoV-2 in convalescent individuals. Nature 584, 437–442 (2020).">3</a></span> (for comparisons of plasma or monoclonal antibodies from convalescent individuals) or HT1080Ace2 cl14 cells<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 13" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR13" id="ref-link-section-d21789e1963" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Schmidt, F. et al. Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses. J. Exp. Med. 217, e20201181 (2020).">13</a></span> (for analyses involving mutant or variant pseudovirus panels), as indicated, for 48 h after which cells were washed with PBS and lysed with Luciferase Cell Culture Lysis 5× reagent (Promega). Nanoluc luciferase activity in lysates was measured using the Nano-Glo Luciferase Assay System (Promega) with the Glomax Navigator (Promega). The obtained relative luminescence units were normalized to those derived from cells infected with SARS-CoV-2 pseudotyped virus in the absence of plasma or monoclonal antibodies. The NT<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">50</span> or half-maximal and 90% inhibitory concentrations for monoclonal antibodies (IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">50</span> and IC<span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">90</span>) were determined using four-parameter nonlinear regression (least squares regression method without weighting; constraints: top, 1; bottom, 0) (GraphPad Prism).</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec17" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Biotinylation of viral protein for use in flow cytometry</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Purified and Avi-tagged SARS-CoV-2 RBD or SARS-CoV-2 RBD KEN mutant (K417N, E484K, N501Y) was biotinylated using the Biotin–Protein Ligase–BIRA kit according to manufacturer’s instructions (Avidity) as previously described. Ovalbumin (Sigma, A5503-1G) was biotinylated using the EZ-Link Sulfo-NHS-LC-Biotinylation kit according to the manufacturer’s instructions (Thermo Scientific). Biotinylated ovalbumin was conjugated to streptavidin-BV711 (BD biosciences, 563262) and RBD to streptavidin-PE (BD Biosciences, 554061) and streptavidin-AF647 (Biolegend, 405237).</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec18" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Flow cytometry and single-cell sorting</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Single-cell sorting by flow cytometry has previously been described<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 3" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR3" id="ref-link-section-d21789e1999" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Robbiani, D. F. et al. Convergent antibody responses to SARS-CoV-2 in convalescent individuals. Nature 584, 437–442 (2020).">3</a></span>. In brief, peripheral blood mononuclear cells were enriched for B cells by negative selection using a pan-B-cell isolation kit according to the manufacturer’s instructions (Miltenyi Biotec, 130-101-638). The enriched B cells were incubated in FACS buffer (1× PBS, 2% FCS, 1 mM EDTA) with the following anti-human antibodies (all at 1:200 dilution): anti-CD20-PECy7 (BD Biosciences, 335793), anti-CD3-APC-eFluro 780 (Invitrogen, 47-0037-41), anti-CD8-APC-eFluor 780 (Invitrogen, 47-0086-42), anti-CD16-APC-eFluor 780 (Invitrogen, 47-0168-41), anti-CD14-APC-eFluor 780 (Invitrogen, 47-0149-42), as well as Zombie NIR (BioLegend, 423105) and fluorophore-labelled RBD and ovalbumin (Ova) for 30 min on ice. Single CD3<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−</span>CD8<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−</span>CD14<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−</span>CD16<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−</span>CD20<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">+</span>Ova<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−</span>RBD<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−</span>PE<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">+</span>RBD<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−</span>AF647<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">+</span> B cells were sorted into individual wells of 96-well plates containing 4 μl of lysis buffer (0.5 × PBS, 10 mM DTT, 3,000 units per ml RNasin Ribonuclease Inhibitors (Promega, N2615) per well using a FACS Aria III and FACSDiva software (Becton Dickinson) for acquisition and FlowJo for analysis. The sorted cells were frozen on dry ice, and then stored at −80 °C or immediately used for subsequent RNA reverse transcription. For B cell phenotype analysis, in addition to above antibodies, B cells were also stained with following anti-human antibodies: anti- IgG-PECF594 (BD biosciences, 562538), anti-IgM-AF700 (Biolegend, 314538), anti-IgA-Viogreen (Miltenyi Biotec, 130-113-481).</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec19" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Antibody sequencing, cloning and expression</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Antibodies were identified and sequenced as previously described. In brief, RNA from single cells was reverse transcribed (SuperScript III Reverse Transcriptase, Invitrogen, 18080-044) and the cDNA stored at −20 °C or used for subsequent amplification of the variable IGH, IGL and IGK genes by nested PCR and Sanger sequencing. Sequence analysis was performed using MacVector. Amplicons from the first PCR reaction were used as templates for sequence- and ligation-independent cloning into antibody expression vectors. Recombinant monoclonal antibodies were produced and purified as previously described<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 3" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR3" id="ref-link-section-d21789e2036" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Robbiani, D. F. et al. Convergent antibody responses to SARS-CoV-2 in convalescent individuals. Nature 584, 437–442 (2020).">3</a></span>.</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec20" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Biolayer interferometry</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">BLI assays were performed as previously described. In brief, we used the Octet Red instrument (ForteBio) at 30 °C with shaking at 1,000 r.p.m. Epitope-binding assays were performed with protein A biosensor (ForteBio 18-5010), following the manufacturer’s protocol ‘classical sandwich assay’. (1) Sensor check: sensors immersed 30 s in buffer alone (kinetics buffer 10x (ForteBio 18-1105) diluted 1x in PBS1x). (2) Capture first antibody: sensors immersed 10 min with Ab1 at 30 μg ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span>. (3) Baseline: sensors immersed 30 s in buffer alone. (4) Blocking: sensors immersed 5 min with IgG isotype control at 50 μg ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span>. (6) Antigen association: sensors immersed 5 min with RBD at 100 μg ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span>. (7) Baseline: sensors immersed 30 s in buffer alone. (8) Association Ab2: sensors immersed 5 min with Ab2 at 30 μg ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span>. Curve fitting was performed using the Fortebio Octet Data analysis software (ForteBio). Affinity measurement of anti-SARS-CoV-2 IgGs binding were corrected by subtracting the signal obtained from traces performed with IgGs in the absence of WT RBD. The kinetic analysis using protein A biosensor (ForteBio 18-5010) was performed as follows: (1) baseline: 60 s immersion in buffer. (2) loading: 200 s immersion in a solution with 30 μg ml<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">−1</span> IgGs. (3) baseline: 200 s immersion in buffer. (4) Association: 300 s immersion in solution with wild-type RBD at 200, 100, 50 or 25 μg/ml. (5) dissociation: 600 s immersion in buffer. Curve fitting was performed using a fast 1:1 binding model and data analysis software (ForteBio). Mean <i style="box-sizing: inherit;">K</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">D</span> values were determined by averaging all binding curves that matched the theoretical fit with an <i style="box-sizing: inherit;">R</i><span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">2</span> value ≥ 0.8.</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec21" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Plasma antibody avidity assay</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The plasma SARS-CoV-2 antibody avidity assay were performed as previously described.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;"><br /></p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhGUZGImqego5ghFpHaT3KsKG7Zoxusk5r7x6ACXqFlTi_BLeQ8BEr1QdtznBpbnOr3dVD60do7iHIn61NS2-phJ0x-juUvNCo8bIYhXz0Lwmp2NHptyjZsX_Lo_TepQvgQNwenUN_xyjU-/s2048/AdobeStock_282303906.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="1365" data-original-width="2048" height="270" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhGUZGImqego5ghFpHaT3KsKG7Zoxusk5r7x6ACXqFlTi_BLeQ8BEr1QdtznBpbnOr3dVD60do7iHIn61NS2-phJ0x-juUvNCo8bIYhXz0Lwmp2NHptyjZsX_Lo_TepQvgQNwenUN_xyjU-/w583-h270/AdobeStock_282303906.jpeg" width="583" /></a></div><br /><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec22" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><br /></h3><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec22" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><br /></h3><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec22" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Computational analyses of antibody sequences</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Antibody sequences were trimmed based on quality and annotated using Igblastn v.1.14. with IMGT domain delineation system. Annotation was performed systematically using Change-O toolkit v.0.4.540. Heavy and light chains derived from the same cell were paired, and clonotypes were assigned based on their V and J genes using in-house R and Perl scripts (Fig. 2d). All scripts and the data used to process antibody sequences are publicly available on GitHub (<a href="https://github.com/stratust/igpipeline" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">https://github.com/stratust/igpipeline</a>).</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The frequency distributions of human V genes in anti-SARS-CoV-2 antibodies from this study was compared to 131,284,220 IgH and IgL sequences generated by ref. and downloaded from cAb-Rep<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 50" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41586-021-03696-9#ref-CR50" id="ref-link-section-d21789e2112" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Guo, Y., Chen, K., Kwong, P. D., Shapiro, L. & Sheng, Z. cAb-Rep: a database of curated antibody repertoires for exploring antibody diversity and predicting antibody prevalence. Front. Immunol. 10, 2365 (2019).">50</a></span>, a database of human shared BCR clonotypes available at <a href="https://cab-rep.c2b2.columbia.edu/" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">https://cab-rep.c2b2.columbia.edu/</a>. On the basis of the 91 distinct V genes that make up the 6,902 analysed sequences from Ig repertoire of the 10 participants present in this study, we selected the IgH and IgL sequences from the database that are partially coded by the same V genes and counted them according to the constant region. The frequencies shown in Extended Data Fig. 4) are relative to the source and isotype analysed. We used the two-sided binomial test to check whether the number of sequences belonging to a specific IgHV or IgLV gene in the repertoire is different according to the frequency of the same IgV gene in the database. Adjusted <i style="box-sizing: inherit;">P</i> values were calculated using the false discovery rate (FDR) correction. Significant differences are denoted with stars.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Nucleotide somatic hypermutation and CDR3 length were determined using in-house R and Perl scripts. For somatic hypermutations, IGHV and IGLV nucleotide sequences were aligned against their closest germlines using Igblastn and the number of differences were considered nucleotide mutations. The average mutations for V genes were calculated by dividing the sum of all nucleotide mutations across all participants by the number of sequences used for the analysis.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Immunoglobulins grouped into the same clonal lineage had their respective IgH and IgL sequences merged and subsequently aligned, using TranslatorX v.1.1, with the unmutated ancestral sequence obtained from IMGT/V-QUEST reference directory. GCTree (<a href="https://github.com/matsengrp/gctree" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">https://github.com/matsengrp/gctree</a>) was further used to perform the phylogenetic trees construction. Each node represents a unique IgH and IgL combination and the size of each node is proportional to the number of identical sequences. The numbered nodes represent the unobserved ancestral genotypes between the germline sequence and the sequences on the downstream branch.</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec23" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Data presentation</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Figures were arranged in Adobe Illustrator 2020.</p><h3 class="c-article__sub-heading c-article__sub-heading--divider" id="Sec24" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><span style="font-family: Harding, Palatino, serif; font-size: 2.4rem; letter-spacing: -0.01875rem;">Data availability</span></h3></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="91748" data-gtm-vis-first-on-screen-10482319_401="91749" data-gtm-vis-total-visible-time-10482319_393="3900" data-gtm-vis-total-visible-time-10482319_401="3900" data-title="Data availability" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="data-availability-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><div class="c-article-section__content" id="data-availability-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Data are provided in Supplementary Tables 1–8. The raw sequencing data have been deposited at Github (<a href="https://github.com/stratust/igpipeline" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">https://github.com/stratust/igpipeline</a>). This study also uses data from <a href="https://doi.org/10.5061/dryad.35ks2" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">https://doi.org/10.5061/dryad.35ks2</a> and from <a href="https://doi.org/10.1038/s41586-019-0934-8" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">https://doi.org/10.1038/s41586-019-0934-8</a>.</p></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="91968" data-gtm-vis-first-on-screen-10482319_401="91968" data-gtm-vis-total-visible-time-10482319_393="3800" data-gtm-vis-total-visible-time-10482319_401="3800" data-title="Code availability" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="code-availability-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="code-availability" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Code availability</h2><div class="c-article-section__content" id="code-availability-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Computer code to process the antibody sequences and/or associated with Fig. 2 and Extended Data Fig. 5 is available at GitHub (<a href="https://github.com/stratust/igpipeline" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">https://github.com/stratust/igpipeline</a>).</p></div></div></section><div id="MagazineFulltextArticleBodySuffix" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px; padding: 0px;"><section aria-labelledby="Bib1" data-title="References" style="box-sizing: inherit;"><div class="c-article-section" id="Bib1-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Bib1" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">References</h2><div class="c-article-section__content" id="Bib1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><div data-container-section="references" style="box-sizing: inherit; margin: 0px; padding: 0px;"><ol class="c-article-references" style="box-sizing: inherit; list-style: none; margin-bottom: 28px; margin-top: 0px; padding: 0px;"><li class="c-article-references__item js-c-reading-companion-references-item" style="border-bottom: 1px solid rgb(213, 213, 213); box-sizing: inherit; display: flex; flex-wrap: wrap; margin-bottom: 16px; padding-bottom: 16px;"><span class="c-article-references__counter" style="box-sizing: inherit; font-size: 2.4rem; line-height: 1.4; text-align: right;">1.</span><p class="c-article-references__text" id="ref-CR1" style="-webkit-box-flex: 1; box-sizing: inherit; flex: 1 1 0%; margin: 0px 0px 8px; overflow-wrap: break-word; padding: 0px 0px 0px 8px;">Davies, N. 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Exit from germinal center to become quiescent memory B cells depends on metabolic reprograming and provision of a survival signal. <i style="box-sizing: inherit;">J. Exp. Med</i>. <span style="box-sizing: inherit; font-weight: bolder;">218</span>, e20200866 (2021).</p></li></ol><div><br /></div><h1 style="text-align: left;"><a href="https://www.arvigen.com/p/careers.html">CARRIERS</a></h1><div><br /></div><div><br /></div></div></div></div></section></div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-52970684985581422212021-07-01T21:08:00.009-07:002021-07-03T22:01:38.831-07:00Mix-and-match COVID vaccines: the case is growing, but questions remain<p> </p><h1 class="c-article-magazine-title" itemprop="headline" style="-webkit-font-smoothing: antialiased; background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 4.5rem; letter-spacing: min(max(-0.01875rem, 4vw), -0.0625rem); line-height: 1.2; margin: 0px 0px 16px; padding: 0px;"><span style="font-size: 18px; font-weight: normal;">A slew of studies suggests that mixing vaccines provokes potent immune responses, but scientists still want answers on real-world efficacy and rare side effects.</span></h1><div><ul class="c-article-author-list js-etal-collapsed js-no-scroll" data-component-authors-activator="authors-list" data-etal-small="3" data-etal="25" data-test="authors-list" style="background-color: white; box-sizing: inherit; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; list-style: none; margin-bottom: 0px; margin-top: 0px; padding: 0px; width: 751.281px;"><li class="c-author-list__item" style="box-sizing: inherit; display: inline; margin-left: 0px; padding-right: 0px;">Dyani Lewis </li></ul></div><div><br /></div><div><br /></div><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhdknJWBesZuzlcvERjwRERcgcFHCl7ysUFwK8ETATndltnTLRqBxCi78Z2-epJ7RSabRDkdhsyrteiN7gaAytMjpWeuhH_4V-XHNEINkv_0EMsbvc8U654TpoRLjx3PhnvapdrzRgWObr1/s660/gda-covid-19-state-2020-12-20-ok-nard.jpg" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="440" data-original-width="660" height="213" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhdknJWBesZuzlcvERjwRERcgcFHCl7ysUFwK8ETATndltnTLRqBxCi78Z2-epJ7RSabRDkdhsyrteiN7gaAytMjpWeuhH_4V-XHNEINkv_0EMsbvc8U654TpoRLjx3PhnvapdrzRgWObr1/w562-h213/gda-covid-19-state-2020-12-20-ok-nard.jpg" width="562" /></a></div><div class="separator" style="clear: both; text-align: center;"><br /></div><div class="separator" style="clear: both; text-align: center;"><br /></div><div class="separator" style="clear: both; text-align: center;"><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">Mixing COVID-19 vaccines is emerging as a good way to get people the protection they need when faced with safety concerns and unpredictable supplies. Most vaccines against SARS-CoV-2 must be given in two doses, but multiple studies now back up the idea that mixing the Oxford–AstraZeneca jab and the Pfizer–BioNTech vaccine triggers an immune response similar to — or even stronger than — two doses of either vaccine.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">Results announced on Monday by a UK group suggest that the combination sometimes outperforms two shots of the same vaccine, and a similar picture is emerging from German studies</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">People can now “feel a bit more comfortable” with the idea of mix-and-match, says immunologist Leif Erik Sander at Charité University Hospital in Berlin.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">The results are also giving researchers confidence that combining other COVID-19 vaccines, that haven’t yet been tested together, might also work. But at least 16 vaccines have been approved for use in one or more countries, and mix-and-match studies so far have been small, so more extensive trials and long-term monitoring for side effects are sorely needed.</p><p style="background-color: white; box-sizing: inherit; font-family: Harding, Palatino, serif; margin: 0px 0px 28px; padding: 0px; text-align: start;"><a href="https://www.arvigen.com/2021/06/covid-vaccines-and-breastfeeding-what.html"><b><span style="color: red; font-size: x-large;">COVID vaccines and breastfeeding: what the data say</span></b></a><br /></p><h2 style="-webkit-font-smoothing: antialiased; background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.4; margin: 0px 0px 8px; padding: 0px; text-align: start;">Immune system boost</h2><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">Mix-and-match studies were prompted, in large part, by concerns over the safety of the vaccine developed by the University of Oxford and pharmaceutical company AstraZeneca in Cambridge, both in the United Kingdom. The jab has been associated with rare instances of a blood-clotting condition known as thrombosis with thrombocytopaenia — and in March, some European countries decided to halt its use in some groups of people. This left many people partially vaccinated, unless they switched to a different brand for their second dose.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">In May, researchers at the Carlos III Health Institute in Madrid announced results from the CombiVacS trial. The study found a strong immune response in people who were dosed with the vaccine developed by pharmaceutical company Pfizer, based in New York City, and biotechnology firm BioNTech in Mainz, Germany, 8–12 weeks after receiving a dose of the Oxford–AstraZeneca vaccine.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">There was no head-to-head comparison with people who received two doses of the same vaccine, but the authors found that in laboratory tests, those who received the combination produced 37 times more SARS-CoV-2 neutralizing antibodies and 4 times more SARS-CoV-2-specific immune cells, called T cells, than did people who had just one dose of the Oxford–AstraZeneca jab.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">By the end of June, more results had emerged showing a similar effect.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; margin: 0px 0px 28px; padding: 0px; text-align: start;"><span style="font-size: 18px;">Sander and his colleagues looked at 340 health-care workers who had received either two doses of the Pfizer–BioNTech vaccine, or an initial shot of the Oxford–AstraZeneca vaccine followed by a dose of Pfizer–BioNTech. Both regimens triggered an immune response that included neutralizing antibodies and T cells</span><span style="font-size: 13.5px;">.</span></p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">A third study, by researchers at Saarland University in Homburg, Germany, found that the mixed regimen was better at eliciting an immune responses than were two Oxford–AstraZeneca shots. It was also as good as or better than two shots of Pfizer–BioNTech.</p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhFNZMu8ZljK2YWB8V3YFWeJFSK98JBbzj7LeGWgbpcHdODAtrP8-4S7j3l2CINUfRxhQ4q7W3eXPvYmRmh-zUTOwH4kZ0BMqgjmTogV84RvUgjuUlvaqXrKdftFUzxzkAgLtt8fTG25AKz/s750/210413123111YVMT.jpg" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="500" data-original-width="750" height="213" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhFNZMu8ZljK2YWB8V3YFWeJFSK98JBbzj7LeGWgbpcHdODAtrP8-4S7j3l2CINUfRxhQ4q7W3eXPvYmRmh-zUTOwH4kZ0BMqgjmTogV84RvUgjuUlvaqXrKdftFUzxzkAgLtt8fTG25AKz/w404-h213/210413123111YVMT.jpg" width="404" /></a></div><div class="separator" style="clear: both; text-align: center;"><br /></div><div class="separator" style="clear: both; text-align: center;"><br /></div><div class="separator" style="clear: both; text-align: center;"><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">And on 25 June, the team behind the UK trial — known as the Com-COV study — posted a pre-print online showing that a good immune response resulted irrespective of the order in which the two vaccines were given.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">However, the trials so far have been too small to test how effective combinations of vaccines are at preventing people from developing COVID-19. “As long as you don’t have any long-term or any follow-up studies with efficacy calculations, it’s hard to say” the level or duration of protection, says Martina Sester, an immunologist who led the Saarland study.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">Another limitation of the work so far is that there’s no easy way to compare different combinations between studies. Large-scale efficacy studies are becoming more difficult, says Sester. That’s because, as infection rates decrease, the number of people in a study must increase to detect any difference in rates of infection and disease. Trials pitting mix-and-match vaccine sequences against a placebo control would also be unethical, she adds.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">That’s one reason why efforts are under way to determine a ‘correlate of protection’ — a defined level of immune response that confers protection against infection and disease. “This is extremely urgent,” says Sander.</p><h2 style="-webkit-font-smoothing: antialiased; background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.4; margin: 0px 0px 8px; padding: 0px; text-align: start;">A nuanced picture</h2><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">But a nuanced picture is emerging of the magnitude and types of immune response that mixing vaccines produces. And these differences could be exploited to provide the best protection.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">The Oxford–AstraZeneca vaccine uses a harmless virus called an adenovirus to carry genetic material from SARS-CoV-2 into cells. Vaccines using this technology have a good track record of inducing strong T-cell responses, says Sander, whereas vaccines using messenger RNA, such as Pfizer’s, have proved “exceptionally good” at inducing high levels of antibodies.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">Sester says that high levels of antibodies after the second shot are an indicator that the combination approach works. “Neutralizing antibodies are probably a good surrogate for predicting efficacy,” she says, because they help to prevent viral infection. But T cells, especially ‘killer’ T cells that carry a protein called CD8, protect against severe disease by killing cells that have already been infected.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">In the Com-COV study, the highest antibody response was in people receiving the standard two shots of Pfizer–BioNTech, but the response was almost as high in the combination of Oxford–AstraZeneca followed by Pfizer–BioNTech. This combination also had the best T-cell response — more than twice as high as that from the two Pfizer–BioNTech doses.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">Mixing an mRNA vaccine and an adenovirus-based one could therefore provide “the best of two worlds”, Sander explains.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">Sester and her colleagues found subtle differences in T-cell populations depending on the vaccines given. She says that understanding these nuances could lead to individualized strategies. Combinations that provoke good T-cell responses might be better for people who have had organ transplants and are taking medication to suppress their immune systems, for instance, because their bodies will struggle to produce antibodies. “There are many ways of exploiting this knowledge in a strategic way,” she says.</p><h2 style="-webkit-font-smoothing: antialiased; background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.4; margin: 0px 0px 8px; padding: 0px; text-align: start;">Safety concerns remain</h2><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">No mix-and-match trials have yet reported severe side effects. In the Com-COV study, mixing vaccines elicited more side effects than did administering two doses of the same vaccine, according to preliminary data released in May<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a data-action="anchor-link" data-track-category="references" data-track-label="go to reference" data-track="click" href="https://www.nature.com/articles/d41586-021-01805-2#ref-CR5" style="background-color: transparent; box-sizing: inherit; color: #006699; text-decoration-line: none; vertical-align: baseline;">5</a></span>. But this wasn’t the case in the Charité and Saarland studies or CombiVacS, where side effects were no worse than for two shots of the same vaccine.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">That’s probably due to the interval between doses, says Sester. Com-COV participants discussed in the latest paper received their second shot four weeks after the initial dose, whereas participants in the German studies had at least nine weeks between shots. Some Com-COV participants did receive doses at a longer interval; their data are anticipated in July.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">Some safety concerns remain, says Sander. “You’re combining two different vaccines, both of which might have their own profile of adverse events and effects,” he says, which could amplify any problems.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">The studies so far have enrolled only a few hundred people. This means that they are too small to pick up rare events such as the clotting conditions, which according to current estimates occur in around one in 50,000 people after the first Oxford–AstraZeneca vaccine dose and in less than 1 in 1.7 million after the second. The condition has also been associated with an adenovirus vaccine produced by pharmaceutical company Johnson & Johnson in New Brunswick, New Jersey.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">In small studies, “you do not pick up your one-in-1,000 side effect, let alone your one-in-50,000 side effect”, said Matthew Snape, an Oxford vaccine researcher who is leading the Com-COV study, at a press conference on 28 June.</p><h2 style="-webkit-font-smoothing: antialiased; background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.4; margin: 0px 0px 8px; padding: 0px; text-align: start;">The new norm?</h2><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">The lingering possibility of rare side effects is one reason some researchers recommend that people stick to the standard two shots of a single vaccine for now. “To my mind, you are better defaulting to the ones where we know that they work and there’s a known quantity when it comes to their safety,” says Snape.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">But as new variants of SARS-CoV-2 emerge, the results of mix-and-match trials could provide policymakers with the data they need to switch to more protective combinations. “It’s good to have that data in readiness,” says Fiona Russell, a vaccine researcher at the Murdoch Children’s Research Institute in Melbourne, Australia.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">Mix-and-match vaccines could also be used to prevent roll-outs stalling because of supply issues. “If there’s a global shortage of one particular vaccine, then rather than stopping the vaccination programme, it can continue,” says Russell.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">“If it’s an option of either getting a mixed schedule or no second dose, then certainly go for the mixed schedule,” says Snape.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">The Com-COV study has already begun testing other vaccines in people who have received an initial Oxford-AstraZeneca or Pfizer–BioNTech shot. One combination includes the yet-to-be-approved protein-based vaccine developed by the pharmaceutical company Novavax in Gaithersburg, Maryland. Another uses the mRNA vaccine from Moderna in Cambridge, Massachusetts, which has been approved for use in several countries.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;">In the Philippines, a study combining the inactivated-virus vaccine CoronaVac, developed by the company Sinovac in Beijing, with the six other vaccines approved in the country will run until November 2022. And a study by AstraZeneca and the Gamaleya Research Institute in Moscow will test combinations of the Oxford–AstraZeneca jab and Gamaleya’s adenovirus-based Sputnik V shot.</p><p style="background-color: white; box-sizing: inherit; font-family: Harding, Palatino, serif; margin: 0px 0px 28px; padding: 0px; text-align: start;"><span style="color: red; font-size: x-large;"><a href="https://www.arvigen.com/2021/06/natural-selection-increases-female.html"><span style="color: red;"><b>Natural selection increases female fitness by reversing the exaggeration of a male sexually selected trait</b></span></a><br /></span></p><p style="background-color: white; box-sizing: inherit; font-family: Harding, Palatino, serif; margin: 0px 0px 28px; padding: 0px; text-align: start;"><br /></p></div><h2 class="c-article-section__title js-section-title" id="Bib1" style="-webkit-font-smoothing: antialiased; background-color: white; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px; text-align: start;">References</h2><div class="c-article-section__content" id="Bib1-content" style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 40px; padding: 8px 0px 0px; text-align: start;"><div data-container-section="references" style="box-sizing: inherit; margin: 0px; padding: 0px;"><ol class="c-article-references" style="box-sizing: inherit; list-style: none; margin-bottom: 28px; margin-top: 0px; padding: 0px;"><li class="c-article-references__item js-c-reading-companion-references-item" style="border-bottom: 1px solid rgb(213, 213, 213); box-sizing: inherit; display: flex; flex-wrap: wrap; margin-bottom: 16px; padding-bottom: 16px;"><span class="c-article-references__counter" style="box-sizing: inherit; font-size: 2.4rem; line-height: 1.4; text-align: right;">1.</span><p class="c-article-references__text" id="ref-CR1" style="-webkit-box-flex: 1; box-sizing: inherit; flex: 1 1 0%; margin: 0px 0px 8px; padding: 0px 0px 0px 8px;">Liu, X. <i style="box-sizing: inherit;">et al.</i> Preprint at SSRN <a href="https://doi.org/10.2139/ssrn.3874014" style="background-color: transparent; box-sizing: inherit; color: #006699; text-decoration-line: none; vertical-align: baseline;">https://doi.org/10.2139/ssrn.3874014</a> (2021).</p></li><li class="c-article-references__item js-c-reading-companion-references-item" style="border-bottom: 1px solid rgb(213, 213, 213); box-sizing: inherit; display: flex; flex-wrap: wrap; margin-bottom: 16px; padding-bottom: 16px;"><span class="c-article-references__counter" style="box-sizing: inherit; font-size: 2.4rem; line-height: 1.4; text-align: right;">2.</span><p class="c-article-references__text" id="ref-CR2" style="-webkit-box-flex: 1; box-sizing: inherit; flex: 1 1 0%; margin: 0px 0px 8px; padding: 0px 0px 0px 8px;">Hillus, D. <i style="box-sizing: inherit;">et al.</i> Preprint at medRxiv <a href="https://doi.org/10.1101/2021.05.19.21257334" style="background-color: transparent; box-sizing: inherit; color: #006699; text-decoration-line: none; vertical-align: baseline;">https://doi.org/10.1101/2021.05.19.21257334</a> (2021).</p></li><li class="c-article-references__item js-c-reading-companion-references-item" style="border-bottom: 1px solid rgb(213, 213, 213); box-sizing: inherit; display: flex; flex-wrap: wrap; margin-bottom: 16px; padding-bottom: 16px;"><span class="c-article-references__counter" style="box-sizing: inherit; font-size: 2.4rem; line-height: 1.4; text-align: right;">3.</span><p class="c-article-references__text" id="ref-CR3" style="-webkit-box-flex: 1; box-sizing: inherit; flex: 1 1 0%; margin: 0px 0px 8px; padding: 0px 0px 0px 8px;">Schmidt, T. <i style="box-sizing: inherit;">et al.</i> Preprint at medRxiv <a href="https://doi.org/10.1101/2021.06.13.21258859" style="background-color: transparent; box-sizing: inherit; color: #006699; text-decoration-line: none; vertical-align: baseline;">https://doi.org/10.1101/2021.06.13.21258859</a> (2021).</p></li><li class="c-article-references__item js-c-reading-companion-references-item" style="border-bottom: 1px solid rgb(213, 213, 213); box-sizing: inherit; display: flex; flex-wrap: wrap; margin-bottom: 16px; padding-bottom: 16px;"><span class="c-article-references__counter" style="box-sizing: inherit; font-size: 2.4rem; line-height: 1.4; text-align: right;">4.</span><p class="c-article-references__text" id="ref-CR4" style="-webkit-box-flex: 1; box-sizing: inherit; flex: 1 1 0%; margin: 0px 0px 8px; padding: 0px 0px 0px 8px;">Borobia, A. M. <i style="box-sizing: inherit;">et al.</i> Preprint at SSRN <a href="https://doi.org/10.2139/ssrn.3854768" style="background-color: transparent; box-sizing: inherit; color: #006699; text-decoration-line: none; vertical-align: baseline;">https://doi.org/10.2139/ssrn.3854768</a> (2021).</p></li><li class="c-article-references__item js-c-reading-companion-references-item" style="border-bottom: 1px solid rgb(213, 213, 213); box-sizing: inherit; display: flex; flex-wrap: wrap; margin-bottom: 16px; padding-bottom: 16px;"><span class="c-article-references__counter" style="box-sizing: inherit; font-size: 2.4rem; line-height: 1.4; text-align: right;">5.</span><p class="c-article-references__text" id="ref-CR5" style="-webkit-box-flex: 1; box-sizing: inherit; flex: 1 1 0%; margin: 0px 0px 8px; padding: 0px 0px 0px 8px;">Shaw, R. H. <i style="box-sizing: inherit;">et al.</i> <i style="box-sizing: inherit;">Lancet</i> <span style="box-sizing: inherit; font-weight: bolder;">397</span>, 2043–2046 (2021).</p></li></ol></div></div><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px; text-align: start;"><br /></p></div><div class="separator" style="clear: both; text-align: center;"><br /></div><br /><div><br /></div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-12710060549114293342021-06-23T09:33:00.005-07:002021-06-23T22:04:20.893-07:00COVID vaccines and breastfeeding: what the data say<h2 style="text-align: left;"><b><span style="color: red;">COVID vaccines and breastfeeding: what the data say </span></b></h2><div><span style="color: #cc0000;"><br /></span></div><div><span style="color: #cc0000;">
Shannon Hall</span></div><div><br />
<div class="separator" style="clear: both;"><img alt="" border="0" data-original-height="533" data-original-width="800" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhUL7dBQ6ELK0e2gjYJIKyQP9M8S1xiwdOFeqaAhXuo-P0WHnYaaqN_IEig9tXBkJF0Key__E4YfE3XDMkI37eVDNil0enUPziXEYZbvjvkWtFTMCBfnQdgQeR0BusNc2FUduTvmmFE4XQ5/s320/red-tent.jpg" width="320" /><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhUL7dBQ6ELK0e2gjYJIKyQP9M8S1xiwdOFeqaAhXuo-P0WHnYaaqN_IEig9tXBkJF0Key__E4YfE3XDMkI37eVDNil0enUPziXEYZbvjvkWtFTMCBfnQdgQeR0BusNc2FUduTvmmFE4XQ5/s800/red-tent.jpg" style="display: block; padding: 1em 0px; text-align: center;"><br /></a></div>
The vaccines do not pass through breastmilk, but antibodies do — providing hope that breastfed babies might have some level of protection. </div><div><br /></div><div> Molly Siegel had long awaited a COVID-19 vaccine. As an obstetrician at Massachusetts General Hospital in Boston, she regularly saw pregnant people with COVID-19, and knew that the vaccine was the best way to protect herself, her family and others in her workplace. But with a seven-month-old baby at home who was still breastfeeding, she felt hesitant. </div><div><br /></div><div>Understandably so. Following established norms for clinical trials, pregnant and breastfeeding people were not included in any of the trials for COVID-19 vaccines. So, as health systems around the world began to vaccinate eligible adults, scores of lactating people were left to make their decision in the dark. </div><div><br /></div><div>“I certainly was frustrated that there weren’t studies on the vaccine in pregnant and lactating women — that as a group, they were excluded from the research,” Siegel says. “It made it really hard to know, as both the patient and the provider, how to think about the vaccine.”
Still, Siegel could not see any plausible risk to her breast milk (she knew that COVID-19 vaccines contain no live virus, for instance), and focused on the benefit of protecting herself and everyone around her. So she got the shot. Then, she donated samples of her breast milk to researchers who would analyse its contents in one of the first such studies.</div><div><br /></div><div> Now, thanks to Siegel and other participants, scientists are beginning to understand the effects of COVID-19 vaccines on breast milk, and their preliminary results should come as welcome news to the more than 100 million lactating people across the world.
Scientists have so far looked only at the vaccines made by Pfizer–BioNTech and Moderna, and have not detected the vaccines in breast milk. What they have found are antibodies, produced by mothers in response to inoculations, to the coronavirus SARS-CoV-2.
“We’re really happy to have something good to hang our hat on,” says Stephanie Gaw, a perinatologist at the University of California, San Francisco. “The studies are small, they’re still early, but very positive.” Now, researchers want to know whether those antibodies can provide babies with at least partial protection against COVID-19. </div><h3 style="text-align: left;"><span style="color: red;"><b><br /></b><b>
Vaccine questions</b> </span></h3><div><br /></div><div> Throughout the pandemic, pregnant people and new mothers have been faced with a slew of concerns and questions about the coronavirus.</div><div><br /></div><div> One trend that became clear early on is that pregnant people diagnosed with COVID-19 are more likely to be hospitalized than are those of the same age who are not pregnant. That could be because the body is already working hard — the growing uterus pushes upwards, reducing lung capacity, and the immune system is suppressed so as not to harm the baby. Those factors don’t disappear the day a baby is born. As such, some obstetricians suspect that lactating individuals are also susceptible to severe COVID-19.</div><div><br /></div><div> That conclusion might encourage breastfeeding mothers to get vaccinated, but scientists weren’t sure how they would respond to the vaccines, because little is known about the period of lactation.
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So Kathryn Gray, a maternal–fetal medicine specialist at Brigham and Women’s Hospital in Boston, Massachusetts, and her colleagues decided to test how well the Pfizer–BioNTech and Moderna vaccines work in this group. They recruited 131 participants who were about to receive either vaccine and who were lactating, pregnant or neither, and found that the lactating individuals (which included Siegel and 30 others) generated the same robust antibody response as did those who were not lactating. In other words: the vaccine is just as beneficial for breastfeeding mums. </div><div><br /></div><div> A second study by Gaw and her team, posted on the preprint server medRxiv, agrees2. The team drew blood from 23 lactating participants and found that antibodies to SARS-CoV-2 increased after their second dose. </div><div><br /></div><div> But for many parents, the looming question — as Siegel asked herself — was whether a COVID-19 vaccine would harm a nursing infant. After all, some medications are not recommended during lactation because they pass through breast milk to infants. Nursing mothers are advised against taking high doses of aspirin, for example; even after low doses, mothers are warned to monitor the infant for signs of bruising and bleeding. Some vaccines are off limits, too. The US Centers for Disease Control and Prevention (CDC) advises nursing mothers against receiving the yellow-fever vaccine, which involves a live, weakened form of the virus, on the off-chance that an infection passes to the infant.
Because of such cases, some pharmacists and vaccine administrators have been urging nursing mothers to discard their breast milk after they are vaccinated. </div><div><br /></div><div> I think that clearly shows ignorance and a lack of understanding,” says Kirsi Jarvinen-Seppo, an immunologist at the University of Rochester Medical Center in Rochester, New York. “There seems to be an awful amount of misinformation out there on all levels.</div><div><br /></div><div>”<span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif;">Unlike the yellow-fever vaccine, COVID-19 vaccines do not carry a risk of igniting an active infection. In addition, COVID-19 vaccines are extremely unlikely to cross into breast milk. The fragile messenger RNA used in the Pfizer–BioNTech and Moderna vaccines, for example, is designed to break down so quickly that it should never leave the cells where it was injected — let alone get into the bloodstream and then the breast. In fact, researchers don’t expect that any of the current vaccines will be excreted into breast milk.</span></div><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; margin: 0px 0px 28px; padding: 0px;">To that end, the World Health Organization recommends that mothers continue to breastfeed after vaccination. In addition, the CDC and the UK Joint Committee on Vaccination and Immunisation issued statements shortly after the first vaccines were authorized in both countries. These noted that no safety concerns had been identified from the available data, so lactating people could choose to be vaccinated.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; margin: 0px 0px 28px; padding: 0px;">“It’s sort of a backwards way of recommending it,” argues Christina Chambers, a paediatrician at the University of California, San Diego, and the Rady Children’s Hospital. “The foundation is that there’s no reason to avoid it, which is a dilemma.”</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; margin: 0px 0px 28px; padding: 0px;">So Gaw and her colleagues ran a safety check. In a small study<span style="box-sizing: inherit; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a data-action="anchor-link" data-track-category="references" data-track-label="go to reference" data-track="click" href="https://www.nature.com/articles/d41586-021-01680-x#ref-CR3" style="background-color: transparent; box-sizing: inherit; color: #006699; text-decoration-line: none; vertical-align: baseline;">3</a></span>, her team looked at breast milk samples from six participants up to two days after they received the Pfizer–BioNTech or Moderna vaccine, and found no trace of the mRNA in either case. (The group is now scouring a larger number of milk samples for different components of the vaccine, and expanding their study to include all the available COVID-19 vaccines in the United States.)</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; margin: 0px 0px 28px; padding: 0px;"><br /></p><p style="background-color: white; box-sizing: inherit; font-family: Harding, Palatino, serif; margin: 0px 0px 28px; padding: 0px;"><a href="https://www.arvigen.com/2018/12/biomarker-risk-score-may-identify-women.html"><b><span style="font-size: large;">Biomarker Risk Score May Identify Women at Risk for Gestational Diabetes</span></b></a><br /></p><h2 style="-webkit-font-smoothing: antialiased; background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.4; margin: 0px 0px 8px; padding: 0px;"><span style="box-sizing: inherit; font-weight: bolder;">Liquid gold</span></h2><div><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px;">There is one type of particle that scientists are eager to see in breast milk following a vaccine: COVID-19 antibodies.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px;">Researchers have long known that newborn babies don’t effectively produce antibodies against harmful bacteria and viruses; and it can take three to six months for this kind of protection to kick in. To help in those early days, a mother’s breast milk overflows with antibodies capable of staving off potential threats.</p><p style="background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px 0px 28px; padding: 0px;">“It’s specifically designed by the mother, and by Mother Nature, to provide the child with the child’s first vaccine,” says Hedvig Nordeng at the University of Oslo, who specializes in medication use and safety in pregnancy and lactation. “Breast milk by itself is more than nutrition, breast milk is medicine.”</p></div><div> In the mother, immune cells called B lymphocytes (or B cells) constantly produce antibodies. </div><div><br /></div><div>Then, once lactation begins, the mammary glands send out a chemical signal that draws these B cells to the breast — where they park in the glands and produce thousands of antibodies per second, ready to move into the breast milk in huge quantities. But unlike molecules from medications, coffee and alcoholic drinks, which are so small they can pass into the breast milk on their own (although at diluted levels), antibodies are too large to do so. Instead, receptors on the surface of the milk ducts grab the antibodies and package them in protective, fluid-filled bubbles that allow them to pass safely through the milk-duct cells and into the milk on the other side.</div><div><br /></div><div> “This process is so magical,” says Galit Alter, an immunologist and virologist at Harvard Medical School in Boston, who worked on Gray’s study. </div><div><br /></div><div> What happens once antibodies reach the baby, however, is more mysterious. Antibodies in the breast milk do not make it into a baby’s bloodstream, but coat the mouth, throat and gut before they’re ultimately digested. Nonetheless, these antibodies seem to provide protection. It could be that they work at the body’s entrances to fend off infection before it takes root.
Not all babies are raised on breast milk, but studies have shown that babies who exclusively breastfeed for their first six months have far fewer middle-ear infections than those who are breastfed for a shorter time, or not at all5. They also have a lower risk of respiratory-tract infections. And lactating mothers who receive the influenza vaccine (and therefore transmit those protective antibodies to their infant through breast milk) provide some protection to babies who are too young to receive the shot.
The same could be true for COVID-19 antibodies.</div><div> Early this year, researchers found that breast milk from people who recover from the virus similarly oozes with antibodies. And a smattering of small studies, many not yet peer reviewed, have found antibodies in breast milk from people who received the vaccine (see ‘Breast-milk benefits’). </div><div><br /></div><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhKDsjgGDJMZ2nXMmAXOBxH2CmRYP5n65Sr2QPf3uEP-aCDKbNsxkt-Y9s6Z3b_u1n_kP7Q10nRoJrqyaqu6DNkhAB5WQZExOOIfh2RIEHT7iX9tMtlg4NmLS_JEgfFaEI5zciX96RUCO9K/s751/d41586-021-01680-x_19278554.png" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="615" data-original-width="751" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhKDsjgGDJMZ2nXMmAXOBxH2CmRYP5n65Sr2QPf3uEP-aCDKbNsxkt-Y9s6Z3b_u1n_kP7Q10nRoJrqyaqu6DNkhAB5WQZExOOIfh2RIEHT7iX9tMtlg4NmLS_JEgfFaEI5zciX96RUCO9K/s320/d41586-021-01680-x_19278554.png" width="320" /></a></div><br /><div><br /></div><div><br /></div><div><br /></div><div> When Gray and her colleagues, for example, checked the blood and the breast milk of lactating mothers who had received a COVID-19 vaccine, they found high levels of COVID-19 antibodies in every sample. </div><div><br /></div><div> “It’s very nice after this past year to have a tiny bit of good news,” says paediatric immunologist Bridget Young at the University of Rochester Medical Center.
And it’s a particularly exciting finding given that babies are not currently eligible to receive any of the available vaccines (although both Pfizer–BioNTech and Moderna have started trials of their COVID-19 vaccines in children as young as six months). </div><div><br /></div><div> Whereas COVID-19 is often mild in younger populations, babies less than two years of age who contract the disease are more likely to be hospitalized than older children are8. That’s thought to be because the bronchioles, the passageways that deliver air to the lungs, are much smaller in babies. In addition, babies and children can develop a severe illness known as MIS-C (multisystem inflammatory syndrome in children), in which different parts of the body become inflamed after the child contracts </div><h2 style="text-align: left;"><span style="color: #cc0000;"><br />
Milk mysteries </span></h2><div><span style="color: #cc0000;"><br /></span></div><div><br /></div><div> One of the big unknowns now is how much protection babies receive from breast milk. </div><div><br /></div><div> To begin, scientists aren’t sure whether these antibodies are actually functional — meaning that they would kill the virus that causes COVID-19 if they came into contact with it. But early research is promising. Last year, a team in the Netherlands collected antibodies from the breast milk of people with a previous SARS-CoV-2 infection and found that the samples could neutralize the virus in the laboratory. A month later, Young, Jarvinen-Seppo and their colleagues posted similar findings, which were subsequently published. </div><div><br /></div><div> Both teams are currently conducting the same experiment with vaccine-induced antibodies, following a study by scientists in Israel suggesting that antibodies created after vaccination could stop the virus infecting cells. The authors of that study predict that these antibodies should protect the baby, says Yariv Wine, an immunologist at Tel Aviv University and a co-author of the paper.
But this can happen only if the antibodies persist. Scientists don’t yet know how long vaccinated people will continue to make COVID-19 antibodies, but evidence indicates they do so for a considerable time; one study of 33 people15 suggests that antibody production in adults given the Moderna vaccine continues for at least 6 months. That could mean that babies will continue to receive some protection from their mothers, as long as they continue nursing — although antibody concentrations in breast milk do drop over time. </div><div><br /></div><div> And that constant replenishment is key. Scientists suspect that antibodies are digested in the baby’s gut after hours to days. That means their partial immunity will probably disappear once breastfeeding has ceased. It also suggests that giving breast milk to older children (as many vaccinated mothers have discussed in online forums) probably won’t give them partial immunity — at least not for long.
But even for babies who are exclusively breastfed, clinicians urge mothers to continue to follow public-health strategies when they have visitors. “Anyone who’s handling the baby in close contact really should be vaccinated and should be masked,” says Andrea Edlow, a maternal–fetal medicine specialist at Harvard Medical School and Massachusetts General Hospital, who worked on the study with Gray.</div><div><br /></div><div> Luckily, more data are on the way. Gray and her team will be tracking their participants, including Siegel and others, for a full year (although the details are still being discussed). Gaw’s team at the University of California, San Francisco, is planning to assess the overall health and rate of infections of babies while they’re being breastfed — the million-dollar question at the moment. The two studies pitting vaccine-induced antibodies against the virus in a Petri dish should offer another answer to this question.
Scientists are also working to analyse the antibodies in further detail. Chambers and her colleagues at the University of California, San Diego, for example, currently receive milk samples from roughly 40 participants per day; they also plan to follow the babies’ growth and development.</div><div><br /></div><div> Nonetheless, the results so far are promising enough that most experts would urge nursing mothers to get their shots. “If I had an itty-bitty baby right now, I would not take the risk — I would not wait,” Alter says. “If you can empower your kid with immunity, I wouldn’t even question it.”
</div><div><br /></div><div><a href="https://www.arvigen.com/2021/06/natural-selection-increases-female.html"><b><span style="font-size: large;">Natural selection increases female fitness by reversing the exaggeration of a male sexually selected trait</span></b></a><br /></div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com1tag:blogger.com,1999:blog-204734490966806252.post-45719956503206237102021-06-22T21:57:00.004-07:002021-06-23T10:57:48.782-07:00Natural selection increases female fitness by reversing the exaggeration of a male sexually selected trait<p> </p><h1 class="c-article-title" data-article-title="" data-test="article-title" style="-webkit-font-smoothing: antialiased; background-color: white; box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 3rem; letter-spacing: min(max(-0.01875rem, 4vw), -0.0625rem); line-height: 1.2; margin: 0px 0px 16px; padding: 0px;">Natural selection increases female fitness by reversing the exaggeration of a male sexually selected trait</h1><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi8SsMLJfqDIA4FHgP_bBSw-nMFTfm1BRDgeee1r4wF070DHXE2KA3NTRi07WdEGciZ-Ybj4qKxhu7hiKhrfZW92PllX5SEYDUO8Y-hMC1MxKzlFVlWyxagVjgG0i3JELynj4A6hgHU-Yi3/s750/compressed-jani.jpg" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="500" data-original-width="750" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi8SsMLJfqDIA4FHgP_bBSw-nMFTfm1BRDgeee1r4wF070DHXE2KA3NTRi07WdEGciZ-Ybj4qKxhu7hiKhrfZW92PllX5SEYDUO8Y-hMC1MxKzlFVlWyxagVjgG0i3JELynj4A6hgHU-Yi3/s320/compressed-jani.jpg" width="320" /></a></div><br /><div><br /></div><div><ul class="c-article-author-list js-etal-collapsed js-no-scroll" data-component-authors-activator="authors-list" data-etal-small="3" data-etal="25" data-test="authors-list" style="background-color: white; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; list-style: none; margin-bottom: 0px; margin-top: 0px; padding: 0px; width: 751.281px;"><li class="c-article-author-list__item" style="box-sizing: inherit; color: #222222; display: inline; font-size: 1.6rem; margin-left: 0px; padding-right: 0px;">Kensuke Okada, </li><li class="c-article-author-list__item" style="box-sizing: inherit; color: #222222; display: inline; font-size: 1.6rem; margin-left: 0px; padding-right: 0px;">Masako Katsuki, </li><li class="c-article-author-list__item js-smaller-author-etal" style="box-sizing: inherit; color: #222222; display: inline; font-size: 1.6rem; margin-left: 0px; padding-right: 0px;">Manmohan D. Sharma, </li><li class="c-article-author-list__item js-smaller-author-etal" style="box-sizing: inherit; color: #222222; display: inline; font-size: 1.6rem; margin-left: 0px; padding-right: 0px;">Katsuya Kiyose, </li><li class="c-article-author-list__item js-smaller-author-etal" style="box-sizing: inherit; color: #222222; display: inline; font-size: 1.6rem; margin-left: 0px; padding-right: 0px;">Tomokazu Seko, </li><li class="c-article-author-list__item js-smaller-author-etal" style="box-sizing: inherit; color: #222222; display: inline; font-size: 1.6rem; margin-left: 0px; padding-right: 0px;">Yasukazu Okada, </li><li class="c-article-author-list__item js-smaller-author-etal" style="box-sizing: inherit; color: #222222; display: inline; font-size: 1.6rem; margin-left: 0px; padding-right: 0px;">Alastair J. Wilson & </li><li class="c-article-author-list__item" style="box-sizing: inherit; color: #222222; display: inline; font-size: 1.6rem; margin-left: 0px; padding-right: 0px;">David J. Hosken</li><li class="c-article-author-list__item" style="box-sizing: inherit; display: inline; margin-left: 0px; padding-right: 0px;"><ul class="c-article-author-list js-etal-collapsed js-no-scroll" data-component-authors-activator="authors-list" data-etal-small="3" data-etal="25" data-test="authors-list" style="background-color: white; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; list-style: none; margin-bottom: 0px; margin-top: 0px; padding: 0px; width: 751.281px;"><section aria-labelledby="Abs1" data-gtm-vis-first-on-screen-10482319_393="453" data-gtm-vis-first-on-screen-10482319_401="453" data-gtm-vis-has-fired-10482319_393="1" data-gtm-vis-has-fired-10482319_401="1" data-gtm-vis-total-visible-time-10482319_393="10000" data-gtm-vis-total-visible-time-10482319_401="10000" data-title="Abstract" lang="en" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Abs1-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Abs1" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Abstract</h2><div class="c-article-section__content" id="Abs1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Theory shows how sexual selection can exaggerate male traits beyond naturally selected optima and also how natural selection can ultimately halt trait elaboration. Empirical evidence supports this theory, but to our knowledge, there have been no experimental evolution studies directly testing this logic, and little examination of possible associated effects on female fitness. Here we use experimental evolution of replicate populations of broad-horned flour beetles to test for effects of sex-specific predation on an exaggerated sexually selected male trait (the mandibles), while also testing for effects on female lifetime reproductive success. We find that populations subjected to male-specific predation evolve smaller sexually selected mandibles and this indirectly increases female fitness, seemingly through intersexual genetic correlations we document. Predation solely on females has no effects. Our findings support fundamental theory, but also reveal unforseen outcomes—the indirect effect on females—when natural selection targets sex-limited sexually selected characters.</p></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="19558" data-gtm-vis-first-on-screen-10482319_401="19562" data-gtm-vis-total-visible-time-10482319_393="3900" data-gtm-vis-total-visible-time-10482319_401="3900" data-title="Introduction" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Sec1-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec1" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Introduction</h2><div class="c-article-section__content" id="Sec1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Sexual selection typically acts more strongly on males and is responsible for the evolution of a vast array of exaggerated characters that enhance male sexual fitness components. Lande’s and Kirkpatrick’s models of sexual selection via the Fisher<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">7</span> process—the null models of intersexual selection—clearly shows how this can occur. They also demonstrate how natural selection can oppose sexual selection as trait values move beyond their naturally selected optima . While theory is clear on the joint effects of natural and sexual selection on sexual trait evolution, explicit experimental tests of theoretical predictions are required to fully understand sexual trait evolution.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">One potentially important source of natural selection that could affect the evolution of sexually selected traits is predation, and many studies have shown predation can seemingly oppose the exaggeration of male sexual characters. For example, sexual signals are conspicuous to potential mates but may also attract predators and parasitoid. This is particularly well documented in orthopterans and frogs, and this form of natural selection is probably responsible for the loss of cricket sexual signals on two Hawaiian isl. More generally, predation appears to reverse the evolution of extreme sexually selected phenotypes (reviewed in the ref. ) and males frequently reduce their sexual signaling in response to predation risk, which can result in decreased mating success when risk is high. Nonetheless, while there is ample evidence that predation selects against sexual trait enhancement, there is limited direct experimental verification that this selection causes evolutionary responses in these traits (but see e.g. ).</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Female reproductive success can also be impacted by predation. For example, egg-carrying females can be slower and suffer higher predation rates. Resultant anti-predator behaviors may reduce foraging efficiency and reproductive activity, and thus be costly to females (reviewed in the ref.). Costs can accrue via delayed development, slower growth or postponed reproduction. Nonetheless, while there is ample evidence that predation selects on both females and males, the joint action of selection on the sexes is frequently investigated independently. Unfortunately, without exploring how predation affects both sexes, we are unlikely to fully understand how predation affects sexual trait evolution. This is especially true when intersexual genetic correlations link sexually selected male characters with female fitness, because selection on one sex can affect the other through these correlations. And again, controlled experimental tests for evolutionary responses to predation are usually not undertaken.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Here, we use the broad-horned flour beetle <i style="box-sizing: inherit;">Gnatocerus cornutus</i> to directly test for effects of predation on the evolution of a male sexually selected character (their mandibles) and female lifetime reproductive success (LRS). The enormously enlarged male mandibles are used in male–male fights, and males with larger mandibles have higher fighting and mating success. Females lack this exaggerated character completely. Previous work has shown that males with larger mandibles sire daughters with lower fecundity, and that directly selecting for increased (or decreased) mandible size results in decreased (or increased) female fitness (LRS). This apparently occurs because of the beetle’s genetic architecture, which means evolving larger mandibles results in the correlated evolution of masculinized females (even though females never develop mandibles). Basically, the enlarged male mandible requires a masculinized head and prothorax to function optimally and this means males with larger mandibles have smaller abdomens. Thus although females never develop mandibles, selecting on male mandibles ultimately affects female abdomen size, which likely determines the number of eggs a female carries.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">These previous <i style="box-sizing: inherit;">G. cornutus</i> studies clearly document intralocus sexual conflict over beetle morphology and point to a negative intersexual correlation between (male) mandible size and female fitness, although this link has not been directly established and requires confirmation (c.f.). With this in mind we investigate the beetle’s intersexual genetic architecture for key focal traits using a standard pedigree-based mixed model approach (commonly referred to as an animal model). We also establish replicate (3/treatment) experimental evolution populations subjected to either male or female predation, along with control populations (<i style="box-sizing: inherit;">n</i> = 3, for nine total populations) to investigate how predation affects an exaggerate male trait and female fitness. The assassin bug <i style="box-sizing: inherit;">Amphibolus venator</i>, which preys on various stored-product insect pests including flour beetles, is the model predator. After eight generations of experimental evolution, we measure female fitness (LRS) and a range of morphological characters, including mandible size. Morphology was also measured during experimental evolution. We find strong effects of male-specific predation on morphology and female fitness, while predation on females alone had no effects. We additionally test for associations between male mandible size and predation likelihood and find larger mandibles result in greater likelihood of predation, and also show that evolving under predation made males less effective fighters.</p></div></div></section><section data-gtm-vis-first-on-screen-10482319_393="22905" data-gtm-vis-first-on-screen-10482319_401="22905" data-gtm-vis-total-visible-time-10482319_393="6200" data-gtm-vis-total-visible-time-10482319_401="6200" data-title="Results" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><h3 class="c-article__sub-heading" id="Sec6" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><br /></h3></section><section data-gtm-vis-first-on-screen-10482319_393="22905" data-gtm-vis-first-on-screen-10482319_401="22905" data-gtm-vis-total-visible-time-10482319_393="6200" data-gtm-vis-total-visible-time-10482319_401="6200" data-title="Results" style="box-sizing: inherit; font-family: Harding, Palatino, serif; font-size: 18px;"><h3 class="c-article__sub-heading" id="Sec6" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><a href="https://www.arvigen.com/2018/12/study-of-nature-and-origin-of-squirting.html"><span style="color: red;">Study of Nature and origin of "squirting" in female sexualit</span></a><span style="color: red;">y</span><br /></h3></section><section data-gtm-vis-first-on-screen-10482319_393="22905" data-gtm-vis-first-on-screen-10482319_401="22905" data-gtm-vis-total-visible-time-10482319_393="6200" data-gtm-vis-total-visible-time-10482319_401="6200" data-title="Results" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><h3 class="c-article__sub-heading" id="Sec6" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><br /></h3></section><section data-gtm-vis-first-on-screen-10482319_393="22905" data-gtm-vis-first-on-screen-10482319_401="22905" data-gtm-vis-total-visible-time-10482319_393="6200" data-gtm-vis-total-visible-time-10482319_401="6200" data-title="Results" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Sec2-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec2" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Results</h2><div class="c-article-section__content" id="Sec2-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><h3 class="c-article__sub-heading" id="Sec3" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Genetic architecture</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The genetic parameters estimated from the animal model analyses confirmed what previous experimental evolution studies had only inferred. Likelihood ratio comparisons of univariate models confirmed the presence of additive genetic variance in all four traits (Supplementary Tables 1–2) as expected. Note that the decision to treat the two mass measures (body/abdomen) as the same trait across the sexes was justified by an absence of significant genotype-by-sex (GxS) interactions in univariate models—a GxS implies sex limited genetic variance is present and can manifest as cross-sex genetic correlations (<i style="box-sizing: inherit;">r</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">Gmf</span>) <+1 and/or differences in sex specific additive genetic variances. Here <i style="box-sizing: inherit;">r</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">Gmf</span> was approximately +1 for both traits, but we do note there is a qualitative pattern of higher additive variance for body mass in females (see Supplementary Tables 1 and 2). Multivariate models further confirmed the presence of additive genetic variance (LRT comparison of null model to one with a diagonal genetic matrix; <i style="box-sizing: inherit;">χ</i><span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">2</span><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">4</span> = 74.96, <i style="box-sizing: inherit;">P</i> < 0.0001), as well as significant among-trait genetic correlation structure (LRT comparison of a model with a diagonal genetic matrix with all genetic correlations are fixed to zero to the full model in which all genetic correlations are estimated; <i style="box-sizing: inherit;">χ</i><span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">2</span><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">6</span> = 39.02, <i style="box-sizing: inherit;">P</i> < 0.0001). Parameter estimates from the full multivariate model show substantial genetic variation in all traits measured and reasonably high trait heritability (Table 1). All traits were positively genetically correlated except male mandible size and female fitness (lifetime reproductive success) and male mandible size and abdominal mass, which were both strongly negatively correlation. Individual genetic correlations were nominally significant at <i style="box-sizing: inherit;">α</i> = 0.05 (based on |<i style="box-sizing: inherit;">r</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">G</span> |> 1.96SE) except those between body and abdomen mass and between body mass and female LRS. The lack of correlation between body and abdomen mass is consistent with previous findings.</p><div class="c-article-table" data-container-section="table" data-test="inline-table" id="table-1" style="border: 5px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption class="c-article-table__figcaption" style="box-sizing: inherit; line-height: 1.4; margin-bottom: 16px; word-break: break-word;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41467-021-23804-7/figures/1" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: inline-block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-line: none; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><img alt="figure1" aria-describedby="Fig1" height="1005" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41467-021-23804-7/MediaObjects/41467_2021_23804_Fig1_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></figcaption></figure></div><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-test="figure" data-title="Responses to sex-specific selection through predation over seven generations." id="figure-1" style="border: 5px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-1-desc" style="box-sizing: inherit; font-size: 1.6rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px;">Shown are male mandible size (mm) (<span style="box-sizing: inherit; font-weight: bolder;">a</span>), male abdomen size (mg) (<span style="box-sizing: inherit; font-weight: bolder;">b</span>), male body size (mg) (<span style="box-sizing: inherit; font-weight: bolder;">c</span>), female abdomen size (mg) (<span style="box-sizing: inherit; font-weight: bolder;">d</span>) and female body size (mg) (<span style="box-sizing: inherit; font-weight: bolder;">e</span>) (population means). Black circles are the control populations that were not subjected to selection by predation. Blue squares and red triangles, are the populations with male and female exposure to predators, respectively. Note we did not measure female fitness (lifetime reproductive success: LRS) at every generation as it was not logistically possible. Population means (estimated by measuring 50 male and females per generation), for each population/treatment are shown because only populations evolve and thus population is the biologically relevant unit of replication in an experimental evolution study. Source data are provided as a Source Data file.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /></div></figure></div><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-test="figure" data-title="Trait values after eight generations of experimental evolution." id="figure-2" style="border: 5px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig2" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Fig. 2: Trait values after eight generations of experimental evolution.</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; display: inline-block; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41467-021-23804-7/figures/2" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: inline-block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-line: none; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41467-021-23804-7/MediaObjects/41467_2021_23804_Fig2_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure2" aria-describedby="Fig2" height="936" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41467-021-23804-7/MediaObjects/41467_2021_23804_Fig2_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-2-desc" style="box-sizing: inherit; font-size: 1.6rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px;">Replicate populations (3/treatment) were exposed to either male-only predation (middle columns), female-only predation (right-hand columns) or no predation (controls: left-hand columns), and effects on a range of traits was assessed. Traits were: Male mandible size (mm) (<span style="box-sizing: inherit; font-weight: bolder;">a</span>), Male abdomen size (mg) (<span style="box-sizing: inherit; font-weight: bolder;">b</span>), Male body size (mg) (<span style="box-sizing: inherit; font-weight: bolder;">c</span>), Female lifetime reproductive success (LRS: offspring number) (<span style="box-sizing: inherit; font-weight: bolder;">d</span>), Female abdomen size (mg) (<span style="box-sizing: inherit; font-weight: bolder;">e</span>), and Female body size (mg) (<span style="box-sizing: inherit; font-weight: bolder;">f</span>) (shown are upper and lower quartile (the box) with medians (lines) and each dot represents the mean of one replicate population). Only populations evolve and thus population is the biologically relevant unit of replication in an experimental evolution study. Mean population values were estimated by measuring 20 individual males and female beetles/population. Source data are provided as a Source Data file.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /></div></figure></div><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Predation also affected female LRS (Fig. 2d: <i style="box-sizing: inherit;">F</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2,6</span> = 21.29; <i style="box-sizing: inherit;">P</i> = 0.002) (note females mated with “non-evolved” tester males). Post-hoc <i style="box-sizing: inherit;">t</i>-tests showed that this was because in the male predation treatment female fitness was higher (all <i style="box-sizing: inherit;">P</i> < 0.01), while the other treatments did not differ (control = female predation: <i style="box-sizing: inherit;">P</i> = 0.54). Thus exposing males to predators resulted in the evolution of smaller male mandibles and higher female fitness. As with males, female abdomen size also increased under male predation (Fig. 2e. Overall treatment effect: <i style="box-sizing: inherit;">F</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2,6</span> = 10.75; <i style="box-sizing: inherit;">P</i> = 0.010): post-hoc <i style="box-sizing: inherit;">t-</i>test revealed the male predator exposure group evolved the largest female abdomens (all <i style="box-sizing: inherit;">P</i> ≤ 0.01: control = female predation <i style="box-sizing: inherit;">P</i> = 1.0)). Finally, female body size was unaffected by our experimental regimes (Fig. 2f. <i style="box-sizing: inherit;">F</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">2,6</span> = 1.65; <i style="box-sizing: inherit;">P</i> = 0.269).</p><h3 class="c-article__sub-heading" id="Sec5" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Mandible size and predation likelihood</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">In the direct assessment of associations between mandible size and predation probability, 45 of 70 male <i style="box-sizing: inherit;">G. cornutus</i> were preyed by female <i style="box-sizing: inherit;">A. venator</i> within thirty minutes. The likelihood of being attacked by the predator was positively associated with mandible size (Fig. <a data-track-action="figure anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41467-021-23804-7#Fig3" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">3</a>; Intercept (±se) = −7.18 ± 2.34; <i style="box-sizing: inherit;">χ</i><span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">2</span> = 12.45; <i style="box-sizing: inherit;">P</i> < 0.001. Coefficient (±se) = 19.52 ± 5.89; <i style="box-sizing: inherit;">χ</i><span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">2</span> = 15.09; <i style="box-sizing: inherit;">P</i> < 0.001). Possible reasons for this are discussed below.</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-test="figure" data-title="The association between G. cornutus mandible size (mm) and likelihood of predation by A. venator." id="figure-3" style="border: 5px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig3" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Fig. 3: The association between <i style="box-sizing: inherit;">G. cornutus</i> mandible size (mm) and likelihood of predation by A. <i style="box-sizing: inherit;">venator</i>.</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; display: inline-block; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41467-021-23804-7/figures/3" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: inline-block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-line: none; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41467-021-23804-7/MediaObjects/41467_2021_23804_Fig3_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure3" aria-describedby="Fig3" height="331" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41467-021-23804-7/MediaObjects/41467_2021_23804_Fig3_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-3-desc" style="box-sizing: inherit; font-size: 1.6rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px;">The graph shows that as mandible size increases so does the likelihood of predation. The curve is the fitted frequency from the GLM with dot size representing numbers (1–6) of individual <i style="box-sizing: inherit;">G. cornutus</i> (i.e., larger dots = more beetles). Inset shows an <i style="box-sizing: inherit;">A. venator</i> about to attack a male <i style="box-sizing: inherit;">G. cornutus</i>. Note the size difference. Source data are provided as a Source Data file.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /></div></figure></div><h3 class="c-article__sub-heading" id="Sec6" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhO4RAbIeJ8cGbCPqlQiArRyegogkvV6beiAYvF_cr9BvUr9dfOiYAr5Aq44JXOOLQXuIhzmySg1vpcW0kovjAn1Wtg0-9HH-KQlFXhet7o0cpZeceCAJSgnheoO-oWOY0lYSdhJ0vw5o3J/s2048/origin.jpg" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="1165" data-original-width="2048" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhO4RAbIeJ8cGbCPqlQiArRyegogkvV6beiAYvF_cr9BvUr9dfOiYAr5Aq44JXOOLQXuIhzmySg1vpcW0kovjAn1Wtg0-9HH-KQlFXhet7o0cpZeceCAJSgnheoO-oWOY0lYSdhJ0vw5o3J/s320/origin.jpg" width="320" /></a></div><br />Main results</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">To summarize the main findings: male-specific predation results in the evolution of an altered, more feminized male phenotype that includes reductions in the size of a male-limited sexually selected trait. Additionally, because of the beetle’s genetic architecture, the “new” demasculinized male phenotype is transmitted through to the female phenotype and results in higher fitness females (Fig. 4).</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-test="figure" data-title="The phenotypes after experimental evolution." id="figure-4" style="border: 5px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig4" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Fig. 4: The phenotypes after experimental evolution.</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; display: inline-block; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41467-021-23804-7/figures/4" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: inline-block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-line: none; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41467-021-23804-7/MediaObjects/41467_2021_23804_Fig4_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure4" aria-describedby="Fig4" height="900" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41467-021-23804-7/MediaObjects/41467_2021_23804_Fig4_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-4-desc" style="box-sizing: inherit; font-size: 1.6rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px;">The upper panel is a diagrammatic representation of the male and female phenotypes resulting from male-limited predation in comparison to those resulting from sexual selection on males. Sexual selection (left images) results in enlarged male mandibles, which require a masculinized head and prothorax to operate effectively. This fore-body masculinization results in a smaller male abdomen and because of intersexual correlations for abdomen size, a smaller female abdomen and capacity for fewer eggs, even though females never develop mandibles. Male-limited predation selects against the masculinized phenotype, ultimately resulting in larger male and female abdomens, and hence more eggs and higher fitness females (images on the right). The lower plate shows beetles from the male predation and control treatments and reveals the impact of evolving with male predation described above—both males and female evolve more feminized phenotypes (smaller mandibles for males and larger abdomen for both) compared to the controls (NB controls and female-predation have similar phenotypes (Figs. 1 and 2) hence we show only controls for clarity). Images were created by us.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /></div></figure></div></div></div></section><section data-title="Discussion" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Sec7-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec7" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Discussion</h2><div class="c-article-section__content" id="Sec7-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Predation is frequently invoked as an evolutionary brake on the exaggeration of sexually selected traits and there are many studies consistent with this logic, usually documenting selection against larger characters or assessing macro-evolutionary patterns consistent with it we employ experimental evolution and directly demonstrate that male-specific predation not only reversed the exaggeration of a sexually selected trait, but additionally, this reversal results in higher female fitness. This boost to female fitness occurs because predators select against larger mandibles, and this results in a less masculinized beetle phenotype (including a larger abdomen), and because of shared genetic architecture across the sexes, females also become more feminized and produce more offspring. We discuss these findings in turn.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The likelihood of predation increases with male mandible size and when populations were forced to evolve with predation on males, smaller mandibles evolve. Thus increased natural selection via predation reduces the size of a sexually selected trait, which is broadly consistent with theory (reviewed in the ref. <span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">1</span>). Previous work with <i style="box-sizing: inherit;">G. cornutus</i> suggests why predation has greater impact on males with larger mandibles. Mandible size is negatively phenotypically and genetically associated with locomotor activity, and locomotor activity (running) is a predator escape mechanism in flour beetles. Reduced running and lower escape rates for males with large mandibles would explain the microevolutionary pattern we detect. Interestingly, there is no intersexual correlation for locomotion in this beetle<span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 49" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41467-021-23804-7#ref-CR49" id="ref-link-section-d11114e1318" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Fuchikawa, T. & Okada, K. Inter‐and intra-sexual genetic correlations of exaggerated traits and locomotor activity. J. Evol. Biol. 26, 1979–1987 (2013).">49</a></span>, so there was no expectation that predation on females should impact male running speed and hence morphology. Be that as it may, we clearly show that predation risk increases with mandible size and that this natural selection reverses the evolutionary exaggeration of a sexually selected male trait.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Females from the male-predation populations evolve higher fitness (higher LRS when mating with stock tester males) even though females were not directly exposed to predators. Previously Harano et al. demonstrated that directly artificially selecting for larger (male) mandibles reduced female fitness. This occurred because selection for increased mandible size resulted in a more masculinized phenotype, and Harano et al. suggested this masculinization rippled through inferred intersexually-shared genetic architecture to increase the masculinization of the female phenotype, thereby reducing female fitness. The intersexual genetic associations we document here, especially the negative male mandible-female LRS correlation, flesh out this explanation and confirm previous inference. Negative intersexual fitness associations are common because alleles conferring high fitness to one sex frequently lower fitness in the other. In addition to any sexually antagonistic selection on body morphology, male beetles with larger mandibles are also more aggressive toward females. Thus exposure to males with larger mandibles potentially reduces female LRS due to (misdirected) male attacks. Therefore, there could be two avenues to increased female fitness when predators select against large male mandibles, a reduction in ontogenetic conflict load (intralocus sexual conflict load), and reduced male-to-female aggression. In any case, predation on males causes an evolutionary reduction in mandible size and this results in higher female fitness. Thus male-biased predation indirectly selects for an increase in female quality, which is an interesting outcome.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The net population level effects of sexual selection and sexual conflict over optimal phenotypes are not clear, with for example, evidence that sexual selection can both increase and decrease population extinction rate. Additionally, intralocus sexual conflict (as documented in the flour beetle) is thought to constrain population adaptation because sexually antagonistic selection keeps each sex from its fitness optima. Our results suggest that predator removal of males with the largest sexual traits reduces intralocus (ontogenetic) sexual conflict costs, enhancing female reproductive performance, which should (all else being equal) increase population productivity. Relaxing other sexual conflict can also increase population fitness. It is interesting to note that predators are usually seen as suppressing prey populations, and this can have indirect benefits for ecological competitors of the prey taxon (predation on one species can open up ecological space and lead to higher fitness of the prey’s competitor taxa). As we show, sex-biased predation can have analogous indirect effects intra-specifically (predation on males increases female fitness). However, the indirect impacts we document are unidirectional since female-biased predation does not alter female fitness or male sexual-trait size.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Micro-evolutionary responses to our treatments were not readily apparent until about four or five generations of selection, after which there was clear treatment-specific divergence. Non-linear responses like this are common in selection studies (and see e.g., ) and there are many potential genetic and environmental mechanisms that can generate non-linearity. This includes genetic asymmetries (i.e., allele frequencies being unequal), differentially skewed genetic and environmental variation, non-additive genetic variation, and indirect selection. We have no way of knowing the mechanistic cause of the patterns we document, but note that responses to directly selecting on mandible size previously revealed similar non-linearity. There have also been many previous studies showing that males with smaller mandibles are less competitive and have lower fitness, and that fighting ability is genetically correlated with mandible size. Consistent with these findings, males evolving with male-predation have smaller mandibles and are less adept fighters.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Overall this study provides direct evidence that predator-mediated natural selection can evolutionarily reverse the exaggeration of a sexually selected trait. This finding is consistent with a vast body of fundamental theory and empirical evidence from observational and correlational studies (reviewed in the ref. ). We also reveal interesting outcomes when natural selection targets sex-limited sexually selected characters, since predator removal of a male-imposed conflict load increases female fitness. Thus sex-biased predation within a species can essentially mimic indirect ecological competition effects, as discussed above. Investigating the precise mechanistic detail and population level fitness effects of some of these findings is now required.</p></div></div></section><section data-title="Discussion" style="box-sizing: inherit; font-family: Harding, Palatino, serif;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;"><a href="https://www.arvigen.com/2019/06/infertility-is-painful-journey-to-go.html"><span style="color: red; font-size: large;">Ovulation Induction</span></a><br /></p></section><section data-title="Methods" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><h3 class="c-article__sub-heading" id="Sec12" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><i style="box-sizing: inherit;"><br /></i></h3></section><section data-title="Methods" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><h3 class="c-article__sub-heading" id="Sec12" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><i style="box-sizing: inherit;"><br /></i></h3></section><section data-title="Methods" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="Sec8-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec8" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Methods</h2><div class="c-article-section__content" id="Sec8-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><h3 class="c-article__sub-heading" id="Sec9" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><i style="box-sizing: inherit;">Gnatocerus</i> stock culture</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The <i style="box-sizing: inherit;">G. cornutus</i> beetle culture originated from adults collected in Miyazaki City (31° 54′N, 131° 25′E), Japan, and has been maintained in the laboratory of the National Food Research Institute, Japan, for about 50 years on whole meal enriched with yeast as food. The stock is made up of 1500–2000 beetles per generation and maintained in plastic cups (diameter 95 mm, height 50 mm) with a standing density of between 300 and 400 beetles per cup (for a more detailed description of the stock culture, see ref. ). This beetle is a stored product pest, and thus the laboratory conditions very closely mimic what have become their natural conditions. Virgin males and females were removed from the stock population as final instar larvae. Each larva was placed in one well of a 24-well tissue culture plate (Cellstar; Greiner Bio-One, Frickenhausen, Germany) until adult eclosion because pupation in <i style="box-sizing: inherit;">G. cornutus</i> is inhibited under high larval density. After eclosion, both sexes were allowed to sexually mature for a period of 14 days prior to their use. We performed all rearing and experiments in a chamber maintained at 25 °C, 60% relative humidity, and with a photoperiod cycle of 14:10 h light/dark. All experiments in this study follow this protocol unless stated otherwise.</p><h3 class="c-article__sub-heading" id="Sec10" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">The predator</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The assassin bug <i style="box-sizing: inherit;">Amphibolus venator</i> is predator of stored-product insect pests and preys on various stored-product insect pests including flour beetles. These predators are frequently found in stored product facilities, which are the habitat of <i style="box-sizing: inherit;">G. cornutus</i>(and see Fig. 3). The <i style="box-sizing: inherit;">A. venator</i> culture originated from adults collected in a storehouse in Urasoe City, Okinawa, Japan, and has been maintained in the laboratory for about 5 years. The stock was initiated and maintained at 200 bugs per generation and housed in plastic containers (230 mm × 150 mm × 80 mm) with a standing density of between 30 and 40 bugs per cup. Each nymph was given an excess of food (seven final instar larvae of <i style="box-sizing: inherit;">G. cornutus</i> per week). Each adult female was allowed to mate with a male chosen randomly and to lay eggs in order to maintain the predator stocks. The predatory behavior of <i style="box-sizing: inherit;">A. venator</i> follows a stereotypical sequence: they recognize, chase, and capture prey (here <i style="box-sizing: inherit;">G. cornutus</i>) using their enlarged forelegs (similar to praying mantis)</p><h3 class="c-article__sub-heading" id="Sec11" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><i style="box-sizing: inherit;">Gnatocerus</i> breeding design and estimation of quantitative genetic parameters</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Using a full sib/half sib experimental design, males (sires) (<i style="box-sizing: inherit;">N</i> = 35) were randomly assigned to three virgin females (dams) (all collected from the stock population). Pairs were housed in a plastic container (17 mm diameter, 20 mm high) containing filter paper (17 mm diameter), and successful copulation was indicated by a stable end-to-end connection between the male and female. After mating, dams were immediately removed and individually placed in a plastic cup (70 mm diameter, 25 mm high) containing excess food (20 g). Each female was housed thus for two months to obtain offspring. All offspring from each female were reared to final instar (approximately 8 weeks). Three sons and three daughters per dam per sire (all pairings produced sufficient young) were haphazardly chosen for measurement of male traits (mandible, body, and abdomen size) and female traits (LRS, body, and abdomen size) at 14 days after eclosion (<i style="box-sizing: inherit;">N</i> = 315 per sex) (trait measurement protocols below). Although we were primarily interested in the genetics of male mandible size and female fitness, we included both abdomen size and total body size measures. This is because previous work found that directly selecting for larger mandibles caused a correlated decrease in abdomen size, but had no effect on total body size. This suggests a trade-off between abdomen and prothorax size and sexual conflict over body shape.</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Data from the breeding design were then analysed using pedigree-based animal models fit in ASReml-R. First, to confirm the presence of additive genetic variance in each trait we fit a series of univariate animal models to (male) mandible, body and abdomen size, and to female (LRS). For each trait we compared the model fit to a reduced model with no additive genetic effects using a likelihood ratio test (LRT; adjusted for boundary conditions following). We elected to combine male and female records for both body size and abdomen sizes as additional modeling provided little support for genotype-by-sex interactions (see “Results” section). However, for these traits a fixed effect of sex was included, as well as the (random) additive genetic effect since exploratory analysis showed sexual dimorphism in both traits (body size, males are 0.020 mg (SE 0.005) larger on average, <i style="box-sizing: inherit;">t</i> = 3.58, <i style="box-sizing: inherit;">P</i> < 0.001; abdomen size, males are 0.019 mg (SE 0.008) smaller on average, <i style="box-sizing: inherit;">t</i> = 2.463, <i style="box-sizing: inherit;">P</i> = 0.014)).</p><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">We then fitted a multivariate animal model to estimate genetic correlation (<i style="box-sizing: inherit;">r</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">G</span>) structure among the four traits with fixed effects of sex on body size, abdomen size, as well as their heritability (<i style="box-sizing: inherit;">h</i><span style="box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">2</span>). The residual covariance structure was modeled as an unstructured matrix (but note residual covariance between the sex-limited traits of male mandible size and female LRS is not estimable from the data so was fixed to zero). We also ran reduced multivariate models with i) no genetic effects at all, and ii) a diagonal genetic variance matrix (i.e., genetic variance modeled on all traits but all genetic correlations assumed to equal zero) for comparison to the full model by LRT. This allows statistical inference at the level of the multivariate phenotype. We used estimated standard errors (SE) as a guide to nominal significance of pairwise genetic correlations (assuming approximate 95% CI are given by <i style="box-sizing: inherit;">r</i><span style="bottom: -0.25em; box-sizing: inherit; font-size: 13.5px; line-height: 0; position: relative; vertical-align: baseline;">G</span> ± 1.96SE).</p><h3 class="c-article__sub-heading" id="Sec12" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;"><i style="box-sizing: inherit;">Gnatocerus</i> experimental evolution protocol—sex specific predation</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">We first collected 900 male and 900 female <i style="box-sizing: inherit;">G. cornutus</i> from the stock culture and haphazardly generated nine groups of 100 males and 100 females to establish three male-predation populations, three female-predation populations and three control (no predation) populations (generation 0) (Fig. 5). To simulate predation, 100 males (or females) were housed in a plastic container (150 mm diameter, 50 mm high) containing an excess of beetle food (45 g). Then, five adult female <i style="box-sizing: inherit;">A. venator</i> (20–35 day olds) were randomly collected from the predator culture and placed into the container and the males (females) were exposed to them for two weeks. We then selected ten of the males (females) that survived the 2 weeks to act as sires (dams) of the predation treatments—ten opposite sex individuals were also taken/population to act as the non-selected dams (sires) that were not exposed to predation. We note that survival rate during this predation protocol was approximately 20%. To propagate control populations, ten males and ten females were haphazardly selected per population to act as sires and dams. For each population/treatment the ten males and females were placed in a plastic cup (diameter 95 mm, height 50 mm) with 70 g of medium for 2 months, with males able to mate with females and females were allowed to lay eggs, until final instar larvae were obtained. Final instar larvae were collected to obtain the adults for subsequent generations. When the adults reached 14 days old, 100 males and 100 females per population were randomly selected to (potentially) seed the next generation, and in the predation treatments, exposed to predators as above. We then selected surviving animals as above and repeated for eight generations. Additionally, we also collected 50 males and 50 females per population from generation 1 to 7 to assess mandible, abdomen and body size responses to selection. At generation 8, 20 males and 20 females per population were haphazardly collected for measurement of male traits (mandible, body, and abdomen size) and female traits (LRS, body and abdomen size) (<i style="box-sizing: inherit;">N</i> = 180 per sex) (trait measurement protocols below).</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-test="figure" data-title="A diagrammatic representation of the experimental evolution protocol." id="figure-5" style="border: 5px solid rgb(213, 213, 213); box-sizing: inherit; clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;"><figure style="box-sizing: inherit; margin: 0px;"><figcaption style="box-sizing: inherit;"><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig5" style="box-sizing: inherit; display: block; font-weight: bolder; margin-bottom: 8px; word-break: break-word;">Fig. 5: A diagrammatic representation of the experimental evolution protocol.</span></figcaption><div class="c-article-section__figure-content" style="box-sizing: inherit; margin: 0px 0px 16px; padding: 0px;"><div class="c-article-section__figure-item" style="box-sizing: inherit; display: inline-block; margin: 0px; max-width: 100%; padding: 0px;"><a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41467-021-23804-7/figures/5" rel="nofollow" style="background-color: transparent; box-sizing: inherit; color: #006699; display: inline-block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; text-decoration-line: none; vertical-align: baseline; word-break: break-word;"><picture style="box-sizing: inherit;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41467-021-23804-7/MediaObjects/41467_2021_23804_Fig5_HTML.png?as=webp" style="box-sizing: inherit;" type="image/webp"></source><img alt="figure5" aria-describedby="Fig5" height="969" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41467-021-23804-7/MediaObjects/41467_2021_23804_Fig5_HTML.png" style="border: 0px; box-sizing: inherit; height: auto; max-width: 100%; vertical-align: middle;" width="685" /></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-5-desc" style="box-sizing: inherit; font-size: 1.6rem; margin: 0px; padding: 0px;"><p style="box-sizing: inherit; margin: 0px; overflow-wrap: break-word; padding: 0px;">Included is the fighting assay after eight generations of experimental evolution. <i style="box-sizing: inherit;">N</i> refers to the number of replicate populations/treatment.</p></div></div><div class="u-text-right u-hide-print" style="box-sizing: inherit; margin: 0px; padding: 0px; text-align: right;"><br /></div></figure></div><h3 class="c-article__sub-heading" id="Sec13" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Fighting ability</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">We also conducted male–male fights between evolved males and males from stock populations (Fig. 5). Briefly, after generation eight of experimental evolution, males from the nine populations (control, male predation, and female predation) were collected. We observed 20 contests per population (<i style="box-sizing: inherit;">n</i> = 180 in total) in which focal experimental males competed against a male collected from the stock culture using standard methods. https://pagead2.googlesyndication.com/pagead/js/adsbygoogle.js
In brief, the stock males were marked with white spots [Mitsubishi Paint-Marker] on their elytra for identification. Males 14 days old (after final eclosion) were used for the experiments. Before pairing, males were weighed with the electronic balance. To control for the effect of body size on fighting success, males were paired so that the difference in body size between contestants was less than 0.05 mg. Pairs were placed on filter-paper (17 mm diameter) in a plastic container (17 mm diameter, 20 mm high) and allowed to interact (and fight) for 1 h. Previous work has shown that male fights occur in all trials when staged in this manner. Males that pushed opponents and chased them were denoted the winner. Trials were then continuously observed until fight outcomes could be scored.</p><h3 class="c-article__sub-heading" id="Sec14" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Predation and mandible size</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">To more directly test for possible associations between predation and mandible size, seventy adult female <i style="box-sizing: inherit;">A. venator</i> (20–35 days old) were randomly collected from the stock culture and were individually aspirated into in a plastic dish (90 mm diameter, 10 mm high) lined with a filter paper. After 30 min, one male adult <i style="box-sizing: inherit;">G. cornutus</i> collected from the stock culture (14 days old: mandible length measured prior to exposure) was added to each dish, and subsequent predation behavior was continuously observed for 30 min.</p><h3 class="c-article__sub-heading" id="Sec15" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Male trait measurement</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">We measured overall body mass and the posterior body mass (i.e., mesothorax, metathorax, and abdomen) as an abdomen size indicator (and see comments above. Briefly, each male was frozen at −20 °C immediately after adult emergence. Mass measures were obtained to the nearest 0.01 mg on an electronic balance (Mettler-Toledo AG, Laboratory and Weighing Technologies). The mandible length (±0.01 mm) of each male was measured (±0.01 mm) using a dissecting microscope monitoring system (VM-60; Olympus, Tokyo, Japan). Each specimen was positioned so that its longitudinal and dorsoventral axes were perpendicular to the visual axes of the microscope eyepiece (see ref. for landmarks).</p><h3 class="c-article__sub-heading" id="Sec16" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Female trait measurement</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">To obtain LRS (lifetime reproductive success: our fitness proxy) each female (14 days of post-eclosure) was individually paired with a haphazardly selected male from the stock culture. After copulation, each female was maintained in a plastic cup (70 mm diameter, 25 mm high) containing excess food (20 g) for two months and allowed to lay eggs. This schedule was chosen because most eggs are laid by females within 2 months of mating, and thus this is an accurate index of LRS. To measure the LRS of each female, we counted all adults that emerged in the third month after pairing. After the laying period, each female was frozen at −20 °C. Subsequently, the whole and posterior of the body (body size and abdomen size) were weighed with the electronic balance (as above).</p><h3 class="c-article__sub-heading" id="Sec17" style="-webkit-font-smoothing: antialiased; box-sizing: inherit; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 2.4rem; font-weight: 400; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">Analysis</h3><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">Apart from the genetic parameter estimation with an animal model (using ASReml-R as described above), all analyses were conducted using JMP for Windows version 8. For the experimental evolution, we used population as the unit of replication (=9 DF max.) with single fixed factor (with three levels: the experimental treatments) GLMs for each trait to test for effects of experimental evolution, with post-hoc testing for factor-level differences (note we did not have sufficient DF for a multivariate analysis). Results are as reported even after (conservative) sequential Bonferroni correction. Fighting results were compared (as population rates of winning) using a generalized linear model (GLM) (JMP 7). To directly assess how mandible size affected predation probability we used a GLM with a binomial distribution, a logit-link function, and overdispersion test. Male mandible size was used as the explanatory variable, and predation impact (preyed upon = 1, survive = 0) was the response variable.</p></div></div></section><section data-title="Methods" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div><br /></div></section><section data-title="Methods" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div><br /></div></section><section data-title="Data availability" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px;"><div class="c-article-section" id="data-availability-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="data-availability" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">Data availability</h2><div class="c-article-section__content" id="data-availability-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><p style="box-sizing: inherit; margin: 0px 0px 28px; overflow-wrap: break-word; padding: 0px;">The data that support the findings of this study are provided in Supplementary Data <a data-track-action="supplementary material anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41467-021-23804-7#MOESM3" style="background-color: transparent; box-sizing: inherit; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">1</a>. This includes population mean trait values during and on completion of experimental evolution, fighting data, predation-mandible size data and the pedigree data. Source data are provided with this paper.</p></div></div></section><div id="MagazineFulltextArticleBodySuffix" style="box-sizing: inherit; color: #222222; font-family: Harding, Palatino, serif; font-size: 18px; margin: 0px; padding: 0px;"><section aria-labelledby="Bib1" data-title="References" style="box-sizing: inherit;"><div class="c-article-section" id="Bib1-section" style="box-sizing: inherit; clear: both; margin: 0px; padding: 0px;"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Bib1" style="-webkit-font-smoothing: antialiased; border-bottom: 2px solid rgb(213, 213, 213); box-sizing: inherit; font-size: 2.4rem; letter-spacing: -0.01875rem; line-height: 1.24; margin: 0px; padding: 0px 0px 8px;">References</h2><div class="c-article-section__content" id="Bib1-content" style="box-sizing: inherit; margin: 0px 0px 40px; padding: 8px 0px 0px;"><div data-container-section="references" style="box-sizing: inherit; margin: 0px; padding: 0px;"><ol class="c-article-references" style="box-sizing: inherit; list-style: none; margin-bottom: 28px; margin-top: 0px; padding: 0px;"><li class="c-article-references__item js-c-reading-companion-references-item" style="border-bottom: 1px solid rgb(213, 213, 213); box-sizing: inherit; display: flex; flex-wrap: wrap; margin-bottom: 16px; padding-bottom: 16px;"><span class="c-article-references__counter" style="box-sizing: inherit; font-size: 2.4rem; line-height: 1.4; text-align: right;">1.</span><p class="c-article-references__text" id="ref-CR1" style="-webkit-box-flex: 1; box-sizing: inherit; flex: 1 1 0%; margin: 0px 0px 8px; overflow-wrap: break-word; padding: 0px 0px 0px 8px;">Andersson, M. 1994. <i style="box-sizing: inherit;">Sexual Selection</i> (Princeton University Press, 1994).</p></li><li class="c-article-references__item js-c-reading-companion-references-item" style="border-bottom: 1px solid rgb(213, 213, 213); box-sizing: inherit; display: flex; flex-wrap: wrap; margin-bottom: 16px; padding-bottom: 16px;"><span class="c-article-references__counter" style="box-sizing: inherit; font-size: 2.4rem; line-height: 1.4; text-align: right;">2.</span><p class="c-article-references__text" id="ref-CR2" style="-webkit-box-flex: 1; box-sizing: inherit; flex: 1 1 0%; margin: 0px 0px 8px; overflow-wrap: break-word; padding: 0px 0px 0px 8px;">Shuster, S. 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Bateson, P.) 67–108 (Cambridge University Press, 1983)</p></li></ol></div></div></div></section></div></ul></li></ul></div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com4tag:blogger.com,1999:blog-204734490966806252.post-20629109734672744602021-05-13T04:28:00.003-07:002021-05-13T04:28:55.199-07:00How the world failed to curb COVID<div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjAhkVB9zohwJaaQhX8TXcae4Gr7sYCUXb6j9OPpln5gqcjPSCOmbf-s8R5npOeyNyMe2IqSjw2F_Bc5QYdrDkYt-ZWtzYZPQWILpLqv-rLAI_1ngEz9aFwS1qBrhJVVk6PfMt5s998GUXU/s734/images+%25283%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="418" data-original-width="734" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjAhkVB9zohwJaaQhX8TXcae4Gr7sYCUXb6j9OPpln5gqcjPSCOmbf-s8R5npOeyNyMe2IqSjw2F_Bc5QYdrDkYt-ZWtzYZPQWILpLqv-rLAI_1ngEz9aFwS1qBrhJVVk6PfMt5s998GUXU/s320/images+%25283%2529.jpeg" width="320" /></a></div><br /><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiPu2SMxDSgtsODUO_fAHT8aU1_rOXlTu9pZBS8PSww3BQ17LOeRXFTK0qSRGjzoGKS2tHCRdPHScHenZI0KGBshkN252YpOtK9VLbbhhn08DXZo_odtMKhri2lt7ShUSnitX1IrSdMk1g6/s679/images+%25284%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="452" data-original-width="679" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiPu2SMxDSgtsODUO_fAHT8aU1_rOXlTu9pZBS8PSww3BQ17LOeRXFTK0qSRGjzoGKS2tHCRdPHScHenZI0KGBshkN252YpOtK9VLbbhhn08DXZo_odtMKhri2lt7ShUSnitX1IrSdMk1g6/s320/images+%25284%2529.jpeg" width="320" /></a></div><br /><p><br /></p><p> <span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">Communication breakdown among the World Health Organization, national leaders and the public caused the pandemic to explode in February 2020, investigation says.</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">The World Health Organization (WHO) was too cautious in communicating the risks of COVID-19 early last year, according to the </span>first major investigation<span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;"> of the global pandemic response. Had it been bolder, and had nations heeded its guidance, the pandemic might have been curtailed, say the authors of the report</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">Last year, during the annual World Health Assembly, countries demanded that the WHO initiate an independent review of how the COVID-19 crisis unfolded, in order to draw lessons for the future. The resulting report, released on 12 May, was assembled by a panel of 13 global-health experts partly appointed by, but independent from, the WHO.</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">The lengthy investigation identifies February 2020 as the month when — in a parallel universe — the devastating toll of the pandemic might have been prevented, had countries acted fast to limit the spread of the virus. It goes on to list concrete actions that could help prevent a similar fate should another deadly pathogen emerge, and it lays out a plan for how vaccines can reach low- and middle-income countries as soon as possible, to end the current crisis. “The reality is, we are still in the thick of this,” explains Joanne Liu, a panel member and a health-emergency specialist at McGill University in Montreal, Canada.</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">Some researchers say that the panel’s suggestions on how to strengthen the WHO are too vague. But the panel does succeed in making a few ambitious recommendations, including creating a council of world leaders dedicated to fighting pandemics, says Stephen Morrison, director of global health policy at the Center for Strategic and International Studies in Washington DC. “They are trying to grab a moment that everyone knows will pass pretty fast,” says Morrison.</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;"><br /></span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;"><b>A bolder WHO</b></span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">Early last year, the WHO should have sounded its highest alarm, a ‘public health emergency of international concern’, or PHEIC, about a week earlier than it did on 30 January, the independent panel concluded in a preliminary report. But in its final summary of the investigation, the panel places more emphasis on what happened between that alarm and when the WHO called the crisis a pandemic on 11 March. Unlike in December 2019 and January 2020, by February, the danger of the coronavirus SARS-CoV-2 spreading globally was well known and its toll might have been averted by national containment strategies. “It is glaringly obvious that February 2020 was a lost month,” the report says.</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">A handful of Asian countries made rapid moves in February of last year to curb COVID-19, including instituting comprehensive testing for SARS-CoV-2 and tracking people who tested positive. “But the rest of the world sat on their hands,” says Liu. She and her colleagues assessed how the WHO communicated risk during February 2020, and decided that the agency’s cautious weighing of incomplete evidence may help to account for why many countries failed to take action.</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">When it became obvious that the countries that were wearing masks were faring better than the ones that weren’t,” she says, “the WHO might have said that even though we don’t have all of the data, we should apply the precautionary principle,” and recommend masks. Similarly, the report indicates that governments might have taken the danger of SARS-CoV-2 more seriously had the WHO described the epidemic as a ‘pandemic’ sooner, even though the term is not defined in the agency's protocols for handling health emergencies.</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">Just after the report published, WHO director-general Tedros Adhanom Ghebreyesus announced that he would review the investigation's critiques and proposals, along with two other reports expected in the coming weeks, and discuss reforms with all countries comprising the WHO.</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">Global health experts have long worried that the WHO faces severe limitations in triggering action. It lacks legal power to enforce recommendations and demand information. And it struggles to criticize a government’s actions because it is chronically underfunded and reliant on donations from its member countries and territories. So the panel recommends a higher budget for the agency, and it says that every country with an epidemic must permit WHO officials access tooutbreak locations on short notice — a swipe at the weeks of negotiation required for the first WHO visit to Wuhan, China, in February.</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">Jennifer Nuzzo, an epidemiologist at Johns Hopkins University in Baltimore, Maryland, says these potential changes would be fine, but that they don’t fully address problems mentioned in the report. For instance, countries would need to agree to reform the regulations dictating the WHO’s protocols so that it has the authority to declare a pandemic. Currently, it can only declare a PHEIC. Nuzzo says, “The WHO is only what we deem it to be.”</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;"><b>Preventing future pandemics</b></span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">Among the report’s stronger recommendations is the formation of an organization outside of the WHO — a Global Health Threats Council — to hold countries accountable for curbing pandemics. The council would include the presidents and prime ministers of several high-, middle- and low-income countries, and its role would be to admonish governments if they fail to prepare for, or respond to, health emergencies, based on advice from science agencies. It could be especially powerful if it’s enacted along with a pandemic treaty currently being pushed by European countries, in which governments have pledged to strengthen their responses. “Not a bad idea,” says Morrison, “but I don’t know if any of this is feasible in our deeply divided, nationalist world.</span><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">”</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">For such a council to exist, a diverse and large number of governments would need to lobby the United Nations to adopt it. But Morrison says that nations that tend to be cagey with information are unlikely to back a group designed to pressure them into transparency. Nonetheless, Liu says the panel aims high, to match the stakes of preventing another crisis of this scale. “By 2025, we are expecting $22 trillion in losses,” she says. “This pandemic has paralyzed the planet for 18 months — when was the last time that happened?”</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;"><br /></span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">Liu and her colleagues will present the recommendations to world leaders at the Global Health summit next week, and at the World Health Assembly in late May. Their goal is to find countries willing to take the ideas forward so that they can become policy.</span></p>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-79546228069605138282021-04-22T01:32:00.002-07:002021-04-22T01:32:32.614-07:00Antiviral drug Molnupiravir showing promise in trials against coronavirus<p> </p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg4OdpO0cdWKnVY1cJB3TgtssVSi0xfgVmsBUaSkWjOLo32hoLHjos9NJrS2KIump2sfwOtaYYICvozdR7vbUBKxENPYZw3tmw0zLs_2TIATHLMh-4Gt1T2Kz7Ds9wRMdk4-k9QFxL9HEHs/s739/images+%25281%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="415" data-original-width="739" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg4OdpO0cdWKnVY1cJB3TgtssVSi0xfgVmsBUaSkWjOLo32hoLHjos9NJrS2KIump2sfwOtaYYICvozdR7vbUBKxENPYZw3tmw0zLs_2TIATHLMh-4Gt1T2Kz7Ds9wRMdk4-k9QFxL9HEHs/s320/images+%25281%2529.jpeg" width="320" /></a></div><br /><p></p><p>Antiviral drug Molnupiravir showing promise in trials against coronavirus</p><p>Molnupiravir is currently in clinical trials and could be a promising tool against coronavirus.</p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgmIq3G55IfgkE9fANqv7Wavz8TqVMig_8vuWXU6cQ07BXgLu5eCAeZRY17YViP2_WjkVn5eHtt1q6TU_uhSl4gpdrxZSmlOFqNyVSUbfls7ufnajeds5QP_aeZsJV_UIP_Shxm471z3e5L/s505/images+%25282%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="343" data-original-width="505" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgmIq3G55IfgkE9fANqv7Wavz8TqVMig_8vuWXU6cQ07BXgLu5eCAeZRY17YViP2_WjkVn5eHtt1q6TU_uhSl4gpdrxZSmlOFqNyVSUbfls7ufnajeds5QP_aeZsJV_UIP_Shxm471z3e5L/s320/images+%25282%2529.jpeg" width="320" /></a></div><br /><p><br /></p><div class="article__section article__section_type_text utility__text" style="background-color: white; box-sizing: inherit; color: #2e2e2e; font-family: proximanova, sans-serif; font-size: 16px; margin: 15px; max-width: 100%; overflow-wrap: break-word; overflow: auto;"><p style="box-sizing: inherit; margin: 0px;">Pune,</p><p style="box-sizing: inherit; margin: 0px;">— Over the last year, several drugs have either been developed or tested to treat coronavirus. Now there's another that's showing some promise. </p></div><div class="article__section article__section_type_text utility__text" style="background-color: white; box-sizing: inherit; color: #2e2e2e; font-family: proximanova, sans-serif; font-size: 16px; margin: 15px; max-width: 100%; overflow-wrap: break-word; overflow: auto;"><p style="box-sizing: inherit; margin: 0px;"> Molnupiravir is an antiviral drug in clinical trials.</p></div><div class="article__section article__section_type_text utility__text" style="background-color: white; box-sizing: inherit; color: #2e2e2e; font-family: proximanova, sans-serif; font-size: 16px; margin: 15px; max-width: 100%; overflow-wrap: break-word; overflow: auto;"><p style="box-sizing: inherit; margin: 0px;">We talked with molecular epidemiologist Dr. Jill Roberts about it being a treatment for COVID-19. </p><p style="box-sizing: inherit; margin: 0px;"><br /></p><p style="box-sizing: inherit; margin: 0px;">She says the way that it works is it messes up viral replication. It's shown some use against other viruses such as SARS and MERS.</p><p style="box-sizing: inherit; margin: 0px;"><br /></p><p style="box-sizing: inherit; margin: 0px;">Dr. Roberts says during an animal study on ferrets, they tried to get coronavirus to spread, and it wouldn't. So while it's a treatment designed to prevent hospitalizations and deaths, it also seems to prevent transmission. </p><p style="box-sizing: inherit; margin: 0px;"><br /></p><p style="box-sizing: inherit; margin: 0px;">While there is no timetable about when this would be available, it could be promising for future viruses or pandemics. </p><p style="box-sizing: inherit; margin: 0px;"><br /></p><p style="box-sizing: inherit; margin: 0px;">"This is a great tool to have to be able to know for future viruses maybe if we get a mutant that actually circumvents the vaccines, this drug will still work."</p><p style="box-sizing: inherit; margin: 0px;"><br /></p><p style="box-sizing: inherit; margin: 0px;">Dr. Roberts says now that Merck is behind it, it has the resources to do a big enough trial to send to the FDA to get Emergency Use Authorization or even full authorization. </p><p style="box-sizing: inherit; margin: 0px;"><br /></p><p style="box-sizing: inherit; margin: 0px;">A couple of other positives she pointed out with this drug: it can also be used for people who do not want to get a vaccine or don't have the resources to get shots. And, this is a pill, unlike other treatments right now that are IVs and have to be given at a treatment center. </p><p style="box-sizing: inherit; margin: 0px;"><br /></p><p style="box-sizing: inherit; margin: 0px;"><br /></p></div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-78118280300845887842021-04-20T22:58:00.002-07:002021-04-20T22:58:54.387-07:00IISER, UNMC scientists explore Rapamycin as repurposed drug to treat elderly, obese with COVID-19<p> </p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEim-5RTBPnRSn7xtk3Hc4EFneHzrhCJBHFZjyZ3F9ovEw_X-PAHICmkjLR5g5wCfvWtJpOgVLsA2RfWQd9rY-db2fWXFPLdrnaOUZedZCKZ24PcF_D126fYJCFQAf5N1Vm82g-ldOx8M7Nk/s739/images.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="415" data-original-width="739" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEim-5RTBPnRSn7xtk3Hc4EFneHzrhCJBHFZjyZ3F9ovEw_X-PAHICmkjLR5g5wCfvWtJpOgVLsA2RfWQd9rY-db2fWXFPLdrnaOUZedZCKZ24PcF_D126fYJCFQAf5N1Vm82g-ldOx8M7Nk/s320/images.jpeg" width="320" /></a></div><br /><p></p><p><span style="font-family: "Playfair Display", sans-serif; font-size: 15px;">Researchers at the Indian Institute of Science Education and Research (IISER) Bhopal and the University of Nebraska Medical Centre (UNMC) in the US have identified a drug that can be repurposed to treat </span><span class="t-out-span" style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; position: relative;"><a class="covid-tooltip" style="box-sizing: border-box; color: #df5b43; cursor: pointer; display: inline; margin: 0px; outline: 0px; padding: 0px;">COVID-19</a></span><span style="font-family: "Playfair Display", sans-serif; font-size: 15px;"> . According to the team, 'Rapamycin' is currently being used for treating cancer patients and those who have undergone organ transplantation. Its analogues are commonly available in India and abroad. The research was published in a peer-reviewed paper recently in the reputed International Elsevier journal, </span><em style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px;">Chemico Biological Interactions</em><span style="font-family: "Playfair Display", sans-serif; font-size: 15px;">. The researchers showed that the biochemical working of this drug molecule points to its promise in the treatment of </span><span class="t-out-span" style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; position: relative;"><a class="covid-tooltip" style="box-sizing: border-box; color: #df5b43; cursor: pointer; display: inline; margin: 0px; outline: 0px; padding: 0px;">COVID-19</a></span><span style="font-family: "Playfair Display", sans-serif; font-size: 15px;"> .</span></p><p><span style="font-family: "Playfair Display", sans-serif; font-size: 15px;">"The development of a new drug is time-consuming and cannot be relied on as a solution in combating the immediate pandemic. Drug repurposing is an attractive solution, wherein, an existing drug is used to treat another related or unrelated ailment may be tested against COVID-19 ," said Amjad Husain, Principal Scientist, and Chief Executive Officer, Innovation and Incubation Center for Entrepreneurship (IICE), IISER Bhopal.</span></p><p><span style="font-family: "Playfair Display", sans-serif; font-size: 15px;">"Since the repurposed drug has gone through the clinical development process for the treatment of other diseases and has already been tested for toxicity, many steps in preclinical and early clinical development can be avoided and the drug can be directly tested on COVID-19 subjects in phase-II trials," he added.</span></p><p><span style="font-family: "Playfair Display", sans-serif; font-size: 15px;"><br /></span></p><p><span style="font-family: "Playfair Display", sans-serif; font-size: 15px;">An example of such a repurposed drug is Remdesivir, which was originally developed to treat Hepatitis C infection. The drug has shown limited success in treating COVID-19 patients.</span></p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;">Identification of more such drugs is important given the scale of the pandemic. Rapamycin works differently from Remdesivir. While the latter targets the virus itself, Rapamycin targets the host proteins and may resist the infection.</p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;">"Using repurposed drug such as Rapamycin that targets mTOR, a central molecule affecting multiple signalling pathways, may yield a significant clinical benefit for the treatment of <span class="t-out-span" style="box-sizing: border-box; position: relative;"><a class="covid-tooltip" style="box-sizing: border-box; color: #df5b43; cursor: pointer; display: inline; margin: 0px; outline: 0px; padding: 0px;">COVID-19</a></span> ," Husain said.</p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;">According to the team, one of the main challenges in developing antiviral drugs for COVID-19 has been the extensive mutations that the virus undergoes, which makes one antiviral drug ineffective against another mutant, and the development of drug-resistant strains.</p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;"><br /></p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;">"Treatment with drugs such as Rapamycin will not face that problem because it acts on host proteins and not on the virus. Rapamycin inhibits protein synthesis and can also arrest virus replication, irrespective of the type of mutant. At a biochemical level, apart from inhibiting protein synthesis, Rapamycin has been known to inhibit pro-inflammatory cytokines.</p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;"><br /></p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;">"It is known that severe COVID-19 infection results in an increase in inflammatory cytokines in a process known as the 'cytokine storm'. The inhibitory action of Rapamycin towards cytokines also makes it a promising treatment for COVID-19 ," he said.</p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;"><br /></p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;">In addition, Rapamycin is known to reduce obesity through various pathways and this can help in mitigating the severity of COVID-19 effects in obese people.</p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;"><br /></p><p style="box-sizing: border-box; font-family: "Playfair Display", sans-serif; font-size: 15px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 22px; margin: 0px 0px 20px; padding: 0px;">"Rapamycin is also known to induce autophagy, a cellular recycling process that helps in eliminating the damaged proteins and delaying ageing. Given the connection between age and COVID mortality - more fatalities with older people, the anti-ageing properties of Rapamycin can have protective effects against COVID-10 induced morbidities," Hussain said.</p>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-7725554264424681672021-03-22T22:28:00.000-07:002021-03-22T22:28:04.029-07:00Dangerous Hybrid Corona strain ; Rises the concern of Worlds Scientists<p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"> </span></span></p><div class="separator" style="clear: both; text-align: center;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEidOiTOWyHSsXf-YFav7x3u4K5tcTKhfmB6HxnWg0PnRuAFQySkjI9tdpeTV7-PaEkhyXJE73YqVtQP0-cjI0OZ34eVQcxyvMB8PSgRGD8OdnhUgU_zrpA6DG6brY5-Ai7IXcGh4TjJ5rQL/s739/images+%25281%2529+%25289%2529.jpeg" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="415" data-original-width="739" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEidOiTOWyHSsXf-YFav7x3u4K5tcTKhfmB6HxnWg0PnRuAFQySkjI9tdpeTV7-PaEkhyXJE73YqVtQP0-cjI0OZ34eVQcxyvMB8PSgRGD8OdnhUgU_zrpA6DG6brY5-Ai7IXcGh4TjJ5rQL/s320/images+%25281%2529+%25289%2529.jpeg" width="320" /></a></span></span></div><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><br /></span></span><p></p><p><span style="vertical-align: inherit;"></span></p><div class="separator" style="clear: both; text-align: center;"><span style="vertical-align: inherit;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEggCIoweCWk67ej_yr6SKaO7TXEket3ZaA6qzwsCCN_nmVus0R5Nj4PgMvfEnAQuxakRnTYNe3W44hvEYsd2JNDtaP64Ro8BhdmQtLKmCDhwybyqI0ZVS4OeJbiYZsSbRFWSSpVCNfurJs_/s739/images+%25281%2529+%25289%2529.jpeg" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="415" data-original-width="739" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEggCIoweCWk67ej_yr6SKaO7TXEket3ZaA6qzwsCCN_nmVus0R5Nj4PgMvfEnAQuxakRnTYNe3W44hvEYsd2JNDtaP64Ro8BhdmQtLKmCDhwybyqI0ZVS4OeJbiYZsSbRFWSSpVCNfurJs_/s320/images+%25281%2529+%25289%2529.jpeg" width="320" /></a></span></div><span style="vertical-align: inherit;"><br /><span style="vertical-align: inherit;"><br /></span></span><p></p><p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">Pune: While the corona has exploded in the state, now another shocking information has come to light. </span></span></span><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">The threat of hybrid corona virus has increased in the world. </span></span></span><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">It is unknown at this time what he will do after leaving the post. </span></span></span><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">Scientists do not know how dangerous this new virus is.</span></span></span></span></p><p><br /></p><p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">The new hybrid corona virus is currently spreading rapidly around the world. </span></span></span><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">This hybrid virus is born from a combination of two viruses.</span></span></span></span></p><p><br /></p><p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">The virus is transmitted from person to person. </span></span></span><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">Dave Vaninsberg, a scientist at Emory University in the United States, has brought this hybrid virus to the world's attention.</span></span></span></span></p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiFsy-YBAxYCetP5dsrgos5bUsRblRqzuliKiPJ9IWy0sBVKqNuiNRjnQ1W2mmkEriw97x83iGoXi47H8gzl3ioobcOmnNAxERoFOhbAQ-4w30MDjVQHA_8xZRLEikJgfMIpQQUjDwEA02l/s733/images+%25281%2529+%252810%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="418" data-original-width="733" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiFsy-YBAxYCetP5dsrgos5bUsRblRqzuliKiPJ9IWy0sBVKqNuiNRjnQ1W2mmkEriw97x83iGoXi47H8gzl3ioobcOmnNAxERoFOhbAQ-4w30MDjVQHA_8xZRLEikJgfMIpQQUjDwEA02l/s320/images+%25281%2529+%252810%2529.jpeg" width="320" /></a></div><br /><p><br /></p><p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">The hybrid virus is a combination of B.117 in the UK and B.1.429 in California.</span></span></span></span></p><p><br /></p><p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">The virus is currently spreading in Los Angeles, USA. </span></span></span><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">The new virus also has the ability to neutralize antibodies.</span></span></span></span></p><p><br /></p><p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">The spike proteins of the hybrid virus are stronger than those found in Wuhan.</span></span></span></span></p><p><br /></p><p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">It is questionable how effective the current corona vaccine is against the new virus.</span></span></span></span></p><p><br /></p><p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">A new wave of corona has arrived in many countries, not just the United States. </span></span></span><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">The corona exploded in Mumbai and Maharashtra. </span></span></span><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">In this case, the hybrid corona virus is even more worrying.</span></span></span></span></p><p><br /></p><p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">Experts say that since the virus is rapidly changing its structure, vaccine production also needs to change.</span></span></span></span></p><p><br /></p><p><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">The reason why many people get corona infection even after getting vaccinated is because of hybrid corona, isn't it? </span></span></span><span style="vertical-align: inherit;"><span style="vertical-align: inherit;"><span style="vertical-align: inherit;">Such a question is currently present.</span></span></span></span></p><p><br /></p><p><br /></p>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-15117404028366314282021-03-21T21:17:00.004-07:002021-03-21T21:25:33.608-07:00Why Covishield (Corona Vaccine) is less effective in some patients<div><br /></div><div><br /></div>
Hello friends, <div><br /></div><div> I asked all my doctor friends out of curiosity if they have seen Cold Chain Maintenance by taking Covishield Vaccine. When asked by many, Dr. Abhi Annadate said, "No. I sat on a stool like a sensible patient, put it on my arm, vaccinated. I took it and sat in the hall for half an hour, there was no reaction, I took paracetamol tablets, thanked Sister and the person who gave me the pills and walked home. I didn't see what Sister had pierced, where she had put it, where she had taken it out. It doesn't even come to mind.
"This is a very representative answer.
Everyone did the same and<div class="separator" style="clear: both;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgQaPxMtoMv1DfCO92-mgKMgnRVSoTkHzBJA2A0kYGbkIRqKbyhma9le_ntVJxfs0nUP4fV4Yb6R6DtCvrqOVu8yZI6tCt_ttJHTz41nbOpS1E7vmaDZRMuxh4kWWiyc3ahbTiYu6tusmIi/s640/images+%25281%2529+%25286%2529.jpeg" style="display: block; padding: 1em 0px; text-align: center;"><img alt="" border="0" data-original-height="480" data-original-width="640" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgQaPxMtoMv1DfCO92-mgKMgnRVSoTkHzBJA2A0kYGbkIRqKbyhma9le_ntVJxfs0nUP4fV4Yb6R6DtCvrqOVu8yZI6tCt_ttJHTz41nbOpS1E7vmaDZRMuxh4kWWiyc3ahbTiYu6tusmIi/s320/images+%25281%2529+%25286%2529.jpeg" width="320" /></a></div><div class="separator" style="clear: both;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh7mwN2wHCT9YtbsEo30MxUR9p-SRioNLaJApad7W751henD9egU-RnTrtoSKlR2gmw-qbo1YnrO_cmA935gXHlgzMOmLg1toqC4gqFSiz74ecWHNyIDx7HtVbR4KzYl6j4VA_euj6nXjQ_/s549/images+%25281%2529+%25287%2529.jpeg" style="display: block; padding: 1em 0px; text-align: center;"><img alt="" border="0" data-original-height="309" data-original-width="549" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh7mwN2wHCT9YtbsEo30MxUR9p-SRioNLaJApad7W751henD9egU-RnTrtoSKlR2gmw-qbo1YnrO_cmA935gXHlgzMOmLg1toqC4gqFSiz74ecWHNyIDx7HtVbR4KzYl6j4VA_euj6nXjQ_/s320/images+%25281%2529+%25287%2529.jpeg" width="320" /></a></div> there is nothing wrong with that.</div><div><br /></div><div> But I visited two vaccination centers as the responsibility of IMA, YEOLA BR.PRESIDENT. </div><div><br /></div><div> So the following type was found </div><div><br /></div><div>where in one place the vial was in an ice cube but the ice cube contained water instead of ice. In another place, at four o'clock, the vaccine fell on the vaccine vial on the wool table through the west window. In both cases, there was no cold force to hold the vaccine vial. This story is heartbreaking for all of us. So I decided to start an awareness campaign among doctors about this as a representative of IMA. This article </div><div>...........
If the current corona vaccination does not maintain cold chain, this vaccination campaign will be a curse instead of a boon. Because .... </div><div><br /></div><div> 1. The Covishield vaccine is a life saver ... but it is just as fragile. </div><div><br /></div><div>2. It is useful only if its temperature is maintained at 2 to 10 Degree celcius. </div><div><br /></div><div> 3. If the temperature is not maintained, injection of vaccine will be like injection of distilled water and it will be of no use. And Corona will happen again. </div><div><br /></div><div> 4. The journey from the time the vaccine is made in the factory to the time it is injected directly into the patient is very long and goes through many stages. It goes through many people, vehicles, storage refrigerators, injectors.</div><div><br /></div><div> 5. Maintaining its temperature at 2 to 8 degrees during all these journeys means maintaining its cold chain is a very complex and responsible task. </div><div><br /></div><div> 6. There is talk in the media today that some people are getting coronary heart disease even after taking both doses. So for those people, it's a "cold chain that hasn't been maintained." The possibility that the vaccine has been injected cannot be ruled out. </div><div><br /></div><div> 7. Because a simple fridge does not work to store this vaccine. This is because its temperature is constantly fluctuating. Suppose this simple fridge is closed for a long time, e.g. When it is closed overnight, the temperature drops below 2 degrees. Ice is formed and if the fridge is opened, the cold air inside is heavy and it goes down quickly to the feet. As the hot air cools, the latency heat releases 540 calories as the temperature drops by one degree Celsius, so the air inside the fridge is initially heated and the temperature immediately rises to a maximum of 8 degrees. Which means it's about to be the most delusional time of the year, as well. Also, even if the vaccine is accidentally frozen, it loses its potency. A special ILR (Ice Lined Refrigerator) is required for such vaccines. Its structure consists of vertical ice-filled tubes attached to each other. Besides, it is horizontal, so when opened, the heavy cold remains in it and its temperature is stable at 2 to 8 degrees. We all know this. "I don't want to ice the vaccine, I want to keep it next to the ice." </div><div><br /></div><div> 8. There may be such ILRs in government hospitals, but it is important to make sure that there are freezers in the private hospitals that administer the vaccine. If not, private vaccination centers should be closed immediately. </div><div> 9. The journey and handling of the vaccine from this ILR to the patient is very important and responsible. 8. In order to remove the vaccine from the ILR and deliver it to the vaccine center, it has to be dropped in the "Vaccine Carrier" box. So this "vaccine carrier" It is not possible to take a handful of onions and potatoes and put them in a vaccine carrier by placing them in a vaccine bottle which is called "Vail" in medical language as it will come in contact with the body and its temperature will reach 37.5 degrees and the vaccine will be useless. </div><div><br /></div><div> 10. So ... each of these should be picked up one by one with the tweezers already cooled and put in the vaccine carrier. 10. This vaccine carrier should be chilled with an ice pack one hour in advance. Otherwise the vaccine carrier is taken and filled with the vaccine in it, while the air at this room temperature i.e. 28-30 degrees can cause the vaccine to fail by increasing the temperature of the vaccine vial in this vaccine carrier. The vaccine carrier should be tightly closed when the vaccine is filled. </div><div><br /></div><div> 11. Now, at the vaccination center, the vaccine should be taken by the carrier leader in the shade and in an AC vehicle. In any case, the leader of the vehicle should take care that the heat coming from the window of the vehicle does not fall on it. </div><div> 12. Vaccine should be taken out one by one for injecting with the help of pre-cooled tweezers.
</div><div><br /></div><div>13. Keep the extracted vaccine in an ice pack. Instead, in many places, the vaccine vial (bottle) is seen lying on the table outside, or if it is kept in an ice pack, but the ice pack contains ice water, but the new ice pack is not taken, or summer has just started, the window is warming up. If these vaccine vials are seen on that table, then such vaccines will be of no use.
</div><div><br /></div><div>14. The last and most important step where the mistake is made is not to hold the vaccine bottle with all fingers or fists. Because if it is straightforward, the temperature of the vaccine will be the same as the body and the vaccine will be useless. To do this, grab the bottle by the thumb and the finger near the thumb and hold it at the mouth of the bottle where there is a rubber butt (stop rubber) inside and remove the required dose from it and immediately place the bottle in the ice pack. This means that the temperature will be maintained. This is the process. In-depth training is given to all health workers. But due to the work ethic, it is ignored by the actual vaccinator. And if that happens, this drug worth crores of rupees will be wasted and this huge amount of labor will be wasted. To prevent this from happening, mobile teams should be formed to monitor this. They should keep a close eye on this vaccination. </div><div><br /></div><div> Officials say the vaccine has a 70% potential. It really will be. But even though the vaccine has a capacity of 70-80 per cent, which means it cannot provide 100 per cent protection, if a vaccine with a strict cold chain maintenance is injected, the 70 per cent protection that will be given will save lives. There will be no death. But if the vaccine that is not maintained in cold chain is injected, no one will be protected from corana at all .... </div><div> but .... </div><div> but ...... </div><div>People will be under the delusion of getting vaccinated ... they will not wear masks ... stop taking care Will do. And then corona cases will continue to grow at a similarly rapid rate. This orgy of death will continue like this. Instead of getting protection from vaccination, it will be a curse to you. </div><div>This should be noted with good conscience and care. Dr. S. Patil ... President, I. M. A. Yeola </div><div><br /></div><div><br /></div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-8539474245473855312021-03-18T21:53:00.002-07:002021-03-18T21:53:26.908-07:00Study of Dopamine on male and female mice<p><span style="font-size: large;">Specific neurons release dopamine to regulate pain differently in male and female mice.</span></p><p></p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgNqYgeBNZL5ebQqhWMqQblaQTElHtWtVQZ5ZsnxbiW6QoSy5RzJMwpk1I7bYbwEDRdyA-9wE6wgWRyOmQ9fc8dR2sr8zokYgGi0fdrImfudtoQnTSJ51Eoo-Dx4zq4jfHq8RaTARiCiiGK/s661/images+%25281%2529+%25285%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="464" data-original-width="661" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgNqYgeBNZL5ebQqhWMqQblaQTElHtWtVQZ5ZsnxbiW6QoSy5RzJMwpk1I7bYbwEDRdyA-9wE6wgWRyOmQ9fc8dR2sr8zokYgGi0fdrImfudtoQnTSJ51Eoo-Dx4zq4jfHq8RaTARiCiiGK/s320/images+%25281%2529+%25285%2529.jpeg" width="320" /></a></div><br /><span style="font-size: large;"><br /></span><p></p><p><span style="font-size: large;"><br /></span></p><p>Males and females, generally speaking, experience and respond to pain differently, but scientists have yet to understand all the brain circuits involved in these differences. Now, new research from the UNC School of Medicine lab of Thomas Kash, PhD, shows how neurons use dopamine to regulate pain differently in male and female mice.</p><p><br /></p><p>The discovery, published in the journal Neuron, could help the scientific community devise better pain management strategies, particularly for women, who are disproportionally affected by pain throughout their lifespans.</p><p>Dopamine, long known as the brain's pleasure chemical, can actually regulate a wide variety of behaviors. The dopamine neurons that Kash and his lab looked at had previously been shown to be important for both the rewarding properties and the pain-relieving properties of heroin. Beyond this, several studies have shown that these neurons can regulate attention, suggesting a link between drug abuse, pain, and attention.</p><p><br /></p><p>Previously, using male mice, the Kash lab found that dopaminergic neurons played a key role in how opiates dampen pain, likely through the release of dopamine and glutamate. In the new experiments, his lab focused on a neural pathway starting at the midbrain region called the periaqueductal grey, including part of the dorsal raphe.</p><p><br /></p><p>We focused on this neural pathway because our previous work and that of others show that specific neurons release dopamine to regulate pain responses. Unfortunately, that research was done only in male mice. So we decided to look at both male and female mice, and what we found was very surprising."</p><p><br /></p><p><br /></p><p>Dopamine, long known as the brain's pleasure chemical, can actually regulate a wide variety of behaviors. The dopamine neurons that Kash and his lab looked at had previously been shown to be important for both the rewarding properties and the pain-relieving properties of heroin. Beyond this, several studies have shown that these neurons can regulate attention, suggesting a link between drug abuse, pain, and attention.</p><p><br /></p><p>That brain region is involved in behavioral adaptation – how animals learn to respond to their environment. The neurons that make dopamine in that region operate in conjunction with a brain structure called the bed nucleus of the stria terminalis, or BNST, forming a neural pathway.</p><p><br /></p><p>"We found that activating this pathway reduced pain sensitivity in male mice, but made female mice move more, especially in the presence of something capturing their attention," said first author Waylin Yu, PhD, a former graduate student in the Kash lab and current postdoctoral researcher at UC San Francisco. "We think this is because of the different ways males and females respond to pain."</p><p><br /></p><p>In particular, these experiments seem to indicate that dopamine helps males simply not feel as much pain, while in females, dopamine helps the mice focus attention elsewhere while in the presence of pain.</p><p><br /></p><p>More research is needed, but the Kash lab research shows that activating specific neural projections to the BNST reduces acute and persistent inflammatory pain, providing further evidence that dopamine signaling can enhance the blocking of pain stimuli, thus counteracting severe pain.</p><p><br /></p><p>"We hope to investigate how this pathway can regulate more emotional behaviors associated with chronic pain, and then also look at the dynamics of the system, such as how this pathway works in real time during behavior measurements," Kash said. "These neurons are also implicated in the actions of opioids such as morphine, so we plan to investigate that domain, as well."</p><p>Source:</p><p>University of North Carolina School of Medicine</p>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-5138620670536956972021-03-18T04:28:00.000-07:002021-03-18T04:28:01.165-07:00Women Are More Interested In Sex Than You Think, Studies Show<p> </p><h1 class="wsj-article-headline" style="background-color: white; color: var(--color-coal); font-family: var(--font-serif-display); font-size: 40px; line-height: 1em; margin: 0px 10px 6px; padding: 0px;">Women Are More Interested In Sex Than You Think, Studies Show </h1><div><br /></div><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhSCyGMjrcY4MK5RSWL81_Ag3RCpNx4kA8591kCUKSiDcn6m0m5mTbpzGnwnufzQoJ3aFCWQZNI4Y0pv7ZyBfFEP3zW8kxEjpKIy0ANDeWMRF_uyPcDI_5d4iPlNMUHQ5V6Ck1IJaiT75eQ/s678/images.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="452" data-original-width="678" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhSCyGMjrcY4MK5RSWL81_Ag3RCpNx4kA8591kCUKSiDcn6m0m5mTbpzGnwnufzQoJ3aFCWQZNI4Y0pv7ZyBfFEP3zW8kxEjpKIy0ANDeWMRF_uyPcDI_5d4iPlNMUHQ5V6Ck1IJaiT75eQ/s320/images.jpeg" width="320" /></a></div><br /><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgxL28KHQgke59J1ADHoBwSB-c3ilK2QWT60xFUlEo_eVB9KTms2INo8ANeLpW9EBqjoEb9E3FlJoWVyzDSo_r5rT1rycL8Cr3xdkhR7zWRSn5P545BZzurKjCh0JOO_LQckpZ7Fwfve3-M/s496/images+%25281%2529+%25284%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="331" data-original-width="496" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgxL28KHQgke59J1ADHoBwSB-c3ilK2QWT60xFUlEo_eVB9KTms2INo8ANeLpW9EBqjoEb9E3FlJoWVyzDSo_r5rT1rycL8Cr3xdkhR7zWRSn5P545BZzurKjCh0JOO_LQckpZ7Fwfve3-M/s320/images+%25281%2529+%25284%2529.jpeg" width="320" /></a></div><br /><div><br /></div><div>Men underestimate their wife’s or girlfriend’s sexual desire; read her signals</div><div>Rarely are researchers’ findings so satisfying. Women may want more sex than their husbands or partners think.</div><div><br /></div><div>New research by psychologists at the University of Toronto and the University of Western Ontario, published earlier this month in the Journal of Personality and Social Psychology, found that men in long-term relationships often underestimate how often their wives or girlfriends want to be intimate.</div><div><br /></div><div>The research consists of three studies, following a total of 229 long-term couples, most of whom are heterosexual. (The sample of homosexual couples was too small to be statistically significant, the researchers say.) Participants ranged in age from 18 to 68 years old; the couples had been together six years on average, and they reported they had sex an average of one to two times a week.</div><div><br /></div><div>In study one, 44 couples kept a diary for three weeks: Partners reported on their own level of sexual desire each day, as well as their perception of their partner’s level of desire and their level of relationship satisfaction. In study two, 84 couples came into the laboratory once and reported on the general levels of their desire, their perception of their partner’s desire and their happiness in the relationship. And in study three, 101 couples kept a diary for three weeks, reporting on the same three issues. They were also asked to report how motivated they were each day to avoid sexual rejection.</div><div><br /></div><div>MARRIED SEX BY THE NUMBERS</div><div>According to ‘The Social Organization of Sexuality: Sexual Practices in the United States,’ a 1994 University of Chicago study considered the most comprehensive in the field:</div><div><br /></div><div> Almost 80% of married couples have sex a few times a month or more.</div><div> Thirty-two percent of married couples report having sex two to three times a week.</div><div> Forty-seven percent of married couples report having sex a few times a month.</div><div> Less than 10% of married people say their last sexual event lasted an hour or more.</div><div>All three studies showed the same thing: Men consistently underestimated their female partner’s desire, while the women had an accurate read on whether or not their partner was interested in sex. And on the days when the men thought their partner was less sexually interested than she actually was, the women reported being more satisfied in and committed to the relationship.</div><div><br /></div><div>The researchers believe that men underestimate their partner’s desire to avoid sexual rejection. If a man initiates sex and his wife rebuffs him, he may feel bad or resentful and she may feel annoyed. By assuming she isn’t interested and not initiating sex, he avoids this downward spiral. And he also may work harder to entice her, which may explain why she still feels content on those days. “It is better for the relationship for him to under-perceive, because it avoids complacency,” says Amy Muise, a postdoctoral fellow at the University of Toronto.</div><div><br /></div><div>How much sex is “normal”? Almost 80% of married couples have sex a few times a month or more: 32% report having sex two to three times a week; 47% say they have sex a few times a month, according to “The Social Organization of Sexuality: Sexual Practices in the United States,” a 1994 University of Chicago study considered the most comprehensive in the field.</div><div><br /></div><div>Men have a higher sex drive on average, research has found. But in long-term relationships—typically defined as longer than three years—men are equally as likely as women to be the partner with low sexual desire. A June 2015, article in the journal “Current Sexual Health Reports” reviewed 31 research studies on sexual desire and sexual discrepancy and found no gender differences in which partner had the higher sex drive.</div><div><br /></div><div>“The assumption that women are going to be the lower-desire partner needs to be thrown out,” says Kristen Mark, author of the article and director of the sexual health promotion laboratory at the University of Kentucky.</div><div><br /></div><div>There are a number of reasons why a man might underestimate how much sex his female partner wants, psychologists say. Some women don’t feel comfortable initiating sex. Others give up initiating after their cues are ignored or missed repeatedly. And many just don’t send clear enough signals.</div><div><br /></div><div><br /></div><div>“I will see women in my office who will tell their husband: ‘Remember when I was joking about that sex scene in that movie we saw? Well, I was trying to come onto you,’” says Sari Cooper, a sex and marriage therapist in New York City. “He may need something more overt.”</div><div><br /></div><div>The problem of women not communicating well about their desire is more complex than couples think, Ms. Cooper says. The woman may not really know what she wants sexually, so she has trouble communicating her wishes or would feel uncomfortable following through with what she asked for. Or she may know that she is the higher-desire partner and be trying to spare his feelings, so he doesn’t feel pressured or unmanly if he doesn’t want to have sex.</div><div><br /></div><div>(The women in the Toronto study who said they were more satisfied in their relationship on days when their partners underestimated their sex drive are probably happier in general with their sex life than the couples who show up for sex therapy, the researchers say.)</div><div><br /></div><div>So what can a couple do? Communicate—not just about when they want to have sex or what they like, but also about what signals they use to show their desire. They should also talk about what signals they prefer to receive. “It’s important not to initiate sex in a way that is a turn off to your partner,” the University of Toronto’s Dr. Muise says.</div><div><br /></div><div><br /></div><div>When talking with your partner about sex—or anything sensitive—use the word “we” instead of the word “you.” A good start is to say: “This is important to me. How can we create a situation that is comfortable for both of us?” “That way there is no blaming going on,” Ms. Cooper, the sex therapist, says. “The couple is sitting down to solve the problem together.”</div><div><br /></div><div>If you can tell your partner is interested in sex but you aren't in the mood, acknowledge their desire. Explain that you find your partner attractive and would like to be intimate, just not at the moment. And promise to find another time.</div><div><br /></div><div>Consider having sex even if you’re not in the mood. Research shows that people in long-term relationships who do this—it is called showing “sexual communal strength”—are better able to maintain their sexual desire over time.</div><div><br /></div><div>Think about scheduling sex. It doesn’t sound romantic. But it is essentially what newer couples do when they plan a date. A study of strategies women use to sync their desire with their partner’s, conducted by researchers at Indiana University, in Bloomington, Ind., and the University of Kentucky and published in 2013 in the Journal of Sexual Medicine, showed this to be a very effective at boosting couples’ sexual satisfaction.</div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-5996720850199451122021-03-13T01:56:00.001-08:002021-03-13T01:56:09.918-08:00Boy or girl? It's in the father's genes<p> </p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgfA1Kt11RjPni6ECBVodfzzWA2_KcQsnUTK13199zrHtV53N0GoqiMT_IjDLW7nbXL1vTEcAcMP8SdQYuJjXaZ8_TRW1Gn8xEkF4uOCpGn0a8WbA_o48nopJrnrkfDrycJ5JJTwM5iLoCO/s768/pregnancy-scan-get-baby-sex-wrong_147398-b520871.jpg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="574" data-original-width="768" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgfA1Kt11RjPni6ECBVodfzzWA2_KcQsnUTK13199zrHtV53N0GoqiMT_IjDLW7nbXL1vTEcAcMP8SdQYuJjXaZ8_TRW1Gn8xEkF4uOCpGn0a8WbA_o48nopJrnrkfDrycJ5JJTwM5iLoCO/s320/pregnancy-scan-get-baby-sex-wrong_147398-b520871.jpg" width="320" /></a></div><div class="separator" style="clear: both; text-align: center;"><br /></div><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjIdA6G28Wt26sTfU9HGTHMljX19wPdeSv9rLaBF6JiuLezHFdiNTJcrzmeD2wQu1khoFNpqTO9-3kEnxCrKeJnndjBWLKPwwjellS2eQJe_qh5RaLObScX9Y9LOfrTJuyd60iUHKVV5PN1/s1025/baby-gender-via-pixabay.jpg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="1025" data-original-width="683" height="320" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjIdA6G28Wt26sTfU9HGTHMljX19wPdeSv9rLaBF6JiuLezHFdiNTJcrzmeD2wQu1khoFNpqTO9-3kEnxCrKeJnndjBWLKPwwjellS2eQJe_qh5RaLObScX9Y9LOfrTJuyd60iUHKVV5PN1/w303-h320/baby-gender-via-pixabay.jpg" width="303" /></a></div><br /><div class="separator" style="clear: both; text-align: center;"><br /></div><br /><p></p><dt style="background-color: white; border-radius: 0px !important; box-sizing: border-box; clear: left; color: #595959; float: left; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; font-style: italic; line-height: 1.42857; overflow: hidden; padding-bottom: 5px; text-align: right; text-overflow: ellipsis; white-space: nowrap; width: 70px;">Date:</dt><dd id="date_posted" style="background-color: white; border-radius: 0px !important; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-left: 90px; padding-bottom: 5px;">March 12, 2021</dd><dt style="background-color: white; border-radius: 0px !important; box-sizing: border-box; clear: left; color: #595959; float: left; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; font-style: italic; line-height: 1.42857; overflow: hidden; padding-bottom: 5px; text-align: right; text-overflow: ellipsis; white-space: nowrap; width: 70px;">Source:</dt><dd id="source" style="background-color: white; border-radius: 0px !important; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-left: 90px; padding-bottom: 5px;">Newcastle University</dd><dt style="background-color: white; border-radius: 0px !important; box-sizing: border-box; clear: left; color: #595959; float: left; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; font-style: italic; line-height: 1.42857; overflow: hidden; padding-bottom: 5px; text-align: right; text-overflow: ellipsis; white-space: nowrap; width: 70px;">Summary:</dt><dd id="abstract" style="background-color: white; border-radius: 0px !important; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-bottom: 15px; margin-left: 90px; padding-bottom: 5px;">A study of hundreds of years of family trees suggests a man's genes play a role in him having sons or daughters. Men inherit a tendency to have more sons or more daughters from their parents. This means that a man with many brothers is more likely to have sons, while a man with many sisters is more likely to have daughters.</dd><dd id="abstract" style="background-color: white; border-radius: 0px !important; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-bottom: 15px; margin-left: 90px; padding-bottom: 5px;"><br /></dd><dd id="abstract" style="background-color: white; border-radius: 0px !important; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-bottom: 15px; margin-left: 90px; padding-bottom: 5px;"><p class="lead" id="first" style="border-radius: 0px !important; box-sizing: border-box; font-size: 18px; line-height: 1.4; margin: 0px 0px 20px;">A Newcastle University study involving thousands of families is helping prospective parents work out whether they are likely to have sons or daughters.</p><div id="text" style="border-radius: 0px !important; box-sizing: border-box;"><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">The work by Corry Gellatly, a research scientist at the university, has shown that men inherit a tendency to have more sons or more daughters from their parents. This means that a man with many brothers is more likely to have sons, while a man with many sisters is more likely to have daughters.</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">The research involved a study of 927 family trees containing information on 556,387 people from North America and Europe going back to 1600.</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">"The family tree study showed that whether you’re likely to have a boy or a girl is inherited. We now know that men are more likely to have sons if they have more brothers but are more likely to have daughters if they have more sisters. However, in women, you just can’t predict it," Mr Gellatly explains.</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">Men determine the sex of a baby depending on whether their sperm is carrying an X or Y chromosome. An X chromosome combines with the mother’s X chromosome to make a baby girl (XX) and a Y chromosome will combine with the mother’s to make a boy (XY).</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">The Newcastle University study suggests that an as-yet undiscovered gene controls whether a man’s sperm contains more X or more Y chromosomes, which affects the sex of his children. On a larger scale, the number of men with more X sperm compared to the number of men with more Y sperm affects the sex ratio of children born each year.</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;"><strong style="border-radius: 0px !important; box-sizing: border-box;">Sons or daughters?</strong></p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">A gene consists of two parts, known as alleles, one inherited from each parent. In his paper, Mr Gellatly demonstrates that it is likely men carry two different types of allele, which results in three possible combinations in a gene that controls the ratio of X and Y sperm;</p><ul style="border-radius: 0px !important; box-sizing: border-box; margin-bottom: 10px; margin-top: 0px;"><li style="border-radius: 0px !important; box-sizing: border-box;">Men with the first combination, known as mm, produce more Y sperm and have more sons.</li><li style="border-radius: 0px !important; box-sizing: border-box;">The second, known as mf, produce a roughly equal number of X and Y sperm and have an approximately equal number of sons and daughters.</li><li style="border-radius: 0px !important; box-sizing: border-box;">The third, known as ff produce more X sperm and have more daughters.</li></ul><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">“The gene that is passed on from both parents, which causes some men to have more sons and some to have more daughters, may explain why we see the number of men and women roughly balanced in a population. If there are too many males in the population, for example, females will more easily find a mate, so men who have more daughters will pass on more of their genes, causing more females to be born in later generations,” says Newcastle University researcher Mr Gellatly.</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;"><strong style="border-radius: 0px !important; box-sizing: border-box;">More boys born after the wars</strong></p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">In many of the countries that fought in the World Wars, there was a sudden increase in the number of boys born afterwards. The year after World War I ended, an extra two boys were born for every 100 girls in the UK, compared to the year before the war started. The gene, which Mr Gellatly has described in his research, could explain why this happened.</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">As the odds were in favour of men with more sons seeing a son return from the war, those sons were more likely to father boys themselves because they inherited that tendency from their fathers. In contrast, men with more daughters may have lost their only sons in the war and those sons would have been more likely to father girls. This would explain why the men that survived the war were more likely to have male children, which resulted in the boy-baby boom.</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">In most countries, for as long as records have been kept, more boys than girls have been born. In the UK and US, for example, there are currently about 105 males born for every 100 females.</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">It is well-documented that more males die in childhood and before they are old enough to have children. So in the same way that the gene may cause more boys to be born after wars, it may also cause more boys to be born each year.</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;"><strong style="border-radius: 0px !important; box-sizing: border-box;">How does the gene work?</strong></p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">The trees (above) illustrate how the gene works. It is a simplified example, in which men either have only sons, only daughters, or equal numbers of each, though in reality it is less clear cut. It shows that although the gene has no effect in females, they also carry the gene and pass it to their children.</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">In the first family tree (A) the grandfather is mm, so all his children are male. He only passes on the m allele, so his children are more likely to have the mm combination of alleles themselves. As a result, those sons may also have only sons (as shown). The grandsons have the mf combination of alleles, because they inherited an m from their father and an f from their mother. As a result, they have an equal number of sons and daughters (the great grandchildren).</p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;">In the second tree (B) the grandfather is ff, so all his children are female, they have the ff combination of alleles because their father and mother were both ff. One of the female children has her own children with a male who has the mm combination of alleles. That male determines the sex of the children, so the grandchildren are all male. The grandsons have the mf combination of alleles, because they inherited an m from their father and f from their mother. As a result, they have an equal number of sons and daughters (the great-grandchildren).</p></div><hr style="border-bottom: 0px; border-image: initial; border-left: 0px; border-radius: 0px !important; border-right: 0px; border-top-color: rgb(238, 238, 238); border-top-style: solid; box-sizing: content-box; height: 0px; margin-bottom: 20px; margin-top: 20px;" /><div id="story_source" style="border-radius: 0px !important; box-sizing: border-box;"><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;"><strong style="border-radius: 0px !important; box-sizing: border-box;">Story Source:</strong></p><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;"><a href="https://www.ncl.ac.uk/press/articles/archive/2015/08/boyorgirlitsinthefathersgenes.html" rel="nofollow" style="background-color: transparent; border-radius: 0px !important; box-sizing: border-box; color: #004276; text-decoration-line: none;" target="_blank">Materials</a> provided by <a href="http://www.ncl.ac.uk/" rel="nofollow" style="background-color: transparent; border-radius: 0px !important; box-sizing: border-box; color: #004276; text-decoration-line: none;" target="_blank"><strong style="border-radius: 0px !important; box-sizing: border-box;">Newcastle University</strong></a>. <em style="border-radius: 0px !important; box-sizing: border-box;">Note: Content may be edited for style and length.</em></p></div><hr style="border-bottom: 0px; border-image: initial; border-left: 0px; border-radius: 0px !important; border-right: 0px; border-top-color: rgb(238, 238, 238); border-top-style: solid; box-sizing: content-box; height: 0px; margin-bottom: 20px; margin-top: 20px;" /><div id="journal_references" style="border-radius: 0px !important; box-sizing: border-box;"><p style="border-radius: 0px !important; box-sizing: border-box; margin: 0px 0px 10px;"><strong style="border-radius: 0px !important; box-sizing: border-box;">Journal Reference</strong>:</p><ol class="journal" style="border-radius: 0px !important; box-sizing: border-box; margin: 0px; padding-left: 20px;"><li style="border-radius: 0px !important; box-sizing: border-box; margin: 0px; padding-left: 5px;">Gellatly et al. <strong style="border-radius: 0px !important; box-sizing: border-box;">Trends in Population Sex Ratios May be Explained by Changes in the Frequencies of Polymorphic Alleles of a Sex Ratio Gene</strong>. <em style="border-radius: 0px !important; box-sizing: border-box;">Evolutionary Biology</em>, Dec 11, 2008; DOI: <a href="http://dx.doi.org/10.1007/s11692-008-9046-3" rel="nofollow" style="background-color: transparent; border-radius: 0px !important; box-sizing: border-box; color: #004276; text-decoration-line: none;" target="_blank">10.1007/s11692-008-9046-3</a></li></ol></div></dd>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-42013911717816912082021-03-13T01:45:00.002-08:002021-03-13T01:46:24.648-08:00Prediabetes may be linked to worse brain health<p> </p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg4nac68D6p7iNfLK-Tkc-MfXwVohwmuyWtCQNG2Pxpxi-qSMsN_HOPazO1xtHFFKw3xEnZEVN_QlppGb6QKT4Ku3fgQFAywVqLHJXl59dpW8ti-BJel9jDoT-9Mg2pdegXdqVcwTi4QFJM/s770/prediabetes-brain-health-neurosciecnes.jpeg" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="481" data-original-width="770" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg4nac68D6p7iNfLK-Tkc-MfXwVohwmuyWtCQNG2Pxpxi-qSMsN_HOPazO1xtHFFKw3xEnZEVN_QlppGb6QKT4Ku3fgQFAywVqLHJXl59dpW8ti-BJel9jDoT-9Mg2pdegXdqVcwTi4QFJM/s320/prediabetes-brain-health-neurosciecnes.jpeg" width="320" /></a></div><br /><p></p><h2 class="subtitle" id="subtitle" style="background-color: white; border-radius: 0px; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 18px; font-weight: 500; line-height: 1.1; margin: 5px 0px 0px;">People with prediabetes, whose blood sugar levels are higher than normal, may have an increased risk of cognitive decline and vascular dementia, according to a new study led by UCL researchers.</h2><div><br /></div><div><dt style="background-color: white; border-radius: 0px; box-sizing: border-box; clear: left; color: #595959; float: left; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; font-style: italic; line-height: 1.42857; overflow: hidden; padding-bottom: 5px; text-align: right; text-overflow: ellipsis; white-space: nowrap; width: 70px;">Date:</dt><dd id="date_posted" style="background-color: white; border-radius: 0px; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-left: 90px; padding-bottom: 5px;">March' 13, 2021</dd><dt style="background-color: white; border-radius: 0px; box-sizing: border-box; clear: left; color: #595959; float: left; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; font-style: italic; line-height: 1.42857; overflow: hidden; padding-bottom: 5px; text-align: right; text-overflow: ellipsis; white-space: nowrap; width: 70px;">Source:</dt><dd id="source" style="background-color: white; border-radius: 0px; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-left: 90px; padding-bottom: 5px;">University College London</dd><dt style="background-color: white; border-radius: 0px; box-sizing: border-box; clear: left; color: #595959; float: left; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; font-style: italic; line-height: 1.42857; overflow: hidden; padding-bottom: 5px; text-align: right; text-overflow: ellipsis; white-space: nowrap; width: 70px;">Summary:</dt><dd id="abstract" style="background-color: white; border-radius: 0px; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-bottom: 15px; margin-left: 90px; padding-bottom: 5px;">Researchers analyzed data from the UK Biobank of 500,000 people aged 58 years on average, and found that people with higher than normal blood sugar levels were 42% more likely to experience cognitive decline over an average of four years, and were 54% more likely to develop vascular dementia over an average of eight years (although absolute rates of both cognitive decline and dementia were low).</dd><dd id="abstract" style="background-color: white; border-radius: 0px; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-bottom: 15px; margin-left: 90px; padding-bottom: 5px;"><br /></dd><dd id="abstract" style="background-color: white; border-radius: 0px; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-bottom: 15px; margin-left: 90px; padding-bottom: 5px;"><p class="lead" id="first" style="border-radius: 0px; box-sizing: border-box; font-size: 18px; line-height: 1.4; margin: 0px 0px 20px;">For the study, published in the journal <em style="border-radius: 0px; box-sizing: border-box;">Diabetes, Obesity and Metabolism</em>, researchers analysed data from the UK Biobank of 500,000 people aged 58 years on average, and found that people with higher than normal blood sugar levels were 42% more likely to experience cognitive decline over an average of four years, and were 54% more likely to develop vascular dementia over an average of eight years (although absolute rates of both cognitive decline and dementia were low).</p><div id="text" style="border-radius: 0px; box-sizing: border-box;"><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">The associations remained true after other influential factors had been taken into account -- including age, deprivation, smoking, BMI and whether or not participants had cardiovascular disease.</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">People with prediabetes have blood sugar levels that are higher than usual, but not high enough to be diagnosed with type 2 diabetes. It means they are more at risk of developing diabetes. There are an estimated five to seven million people* with prediabetes in the UK.</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">Lead author Dr Victoria Garfield (UCL Institute of Cardiovascular Science and the UCL MRC Unit for Lifelong Health & Ageing) said: "Our research shows a possible link between higher blood sugar levels -- a state often described as 'prediabetes' -- and higher risks of cognitive decline and vascular dementia. As an observational study, it cannot prove higher blood sugar levels cause worsening brain health. However, we believe there is a potential connection that needs to be investigated further.</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">"Previous research has found a link between poorer cognitive outcomes and diabetes but our study is the first to investigate how having blood sugar levels that are relatively high -- but do not yet constitute diabetes -- may affect our brain health."</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">In the study, researchers investigated how different blood sugar levels, or glycaemic states, were associated with performance in cognitive tests over time, dementia diagnoses, and brain structure measured by MRI scans of the brain. Each of these measures were limited to smaller subsets of the Biobank sample (for instance, only 18,809 participants had follow-up cognitive tests).</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">At recruitment all of the UK Biobank participants underwent an HbA1c test, which determines average blood sugar levels over the past two to three months. Participants were divided into five groups on the basis of the results -- "low-normal" level of blood sugar, normoglycaemia (having a normal concentration of sugar in the blood), prediabetes, undiagnosed diabetes and diabetes. A result between 42-48 mmol/mol (6.0-6.5%) was classified as prediabetes.</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">The researchers used data from repeated assessments of visual memory to determine whether participants had cognitive decline or not. Though absolute rates of cognitive decline were low, people with prediabetes and diabetes had a similarly higher likelihood of cognitive decline -- 42% and 39% respectively.</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">Looking at dementia diagnoses, researchers found that prediabetes was associated with a higher likelihood of vascular dementia, a common form of dementia caused by reduced blood flow to the brain, but not Alzheimer's disease. People with diabetes, meanwhile, were three times more likely to develop vascular dementia than people whose blood sugar levels were classified as normal, and more likely to develop Alzheimer's disease.</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">Senior author Professor Nishi Chaturvedi (UCL MRC Unit for Lifelong Health & Ageing) said: "In this relatively young age group, the risks of cognitive decline and of dementia are very low; the excess risks we observe in relation to elevated blood sugar only modestly increase the absolute rates of ill health. Seeing whether these effects persist as people get older, and where absolute rates of disease get higher, will be important.</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">"Our findings also need to be replicated using other datasets. If they are confirmed, they open up questions about the potential benefits of screening for diabetes in the general population and whether we should be intervening earlier."</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">Among 35,418 participants of the UK Biobank study who underwent MRI brain scans, researchers found that prediabetes was associated somewhat with a smaller hippocampus and more strongly associated with having lesions on the brain (white matter hyperintensities, WMHs) -- both associated with age-related cognitive impairment.</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">The researchers said that some of these differences could be explained by elevated blood pressure, as those participants taking antihypertensive medication were likely to have more WMHs and smaller hippocampal volume. Rather than the treatment having an adverse effect on the brain, the researchers said use of such medication might be an indicator of earlier untreated high blood pressure.</p><p style="border-radius: 0px; box-sizing: border-box; margin: 0px 0px 10px;">People with prediabetes can reduce their risk of developing type 2 diabetes by eating a healthy, balanced diet, being more active, and staying at a healthy weight.</p></div><div class="impact-unit-wrapper" style="border-radius: 0px; box-sizing: border-box; margin: 25px 5px 25px 0px;"><div class="pgs-dpg-btn" data-loaded="true" data-pgs-partner-id="sciencedaily" style="border-radius: 0px; box-sizing: border-box;" widget-loading="true"></div></div></dd><dd id="abstract" style="background-color: white; border-radius: 0px; box-sizing: border-box; color: #333333; font-family: "Helvetica Neue", Helvetica, Arial, sans-serif; font-size: 14px; line-height: 1.42857; margin-bottom: 15px; margin-left: 90px; padding-bottom: 5px;"><br /></dd></div>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-18932496471296502612020-09-28T21:44:00.004-07:002020-09-28T21:44:28.649-07:00How COVID-19 can damage the brain<p> <b><span style="font-size: medium;">How COVID-19 can damage the brain</span></b></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">Some people who become ill with the coronavirus develop neurological symptoms. Scientists are struggling to understand why.</span></p><p></p><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh4k8C1BRsrTaK9JUros4RTJ9V18w-HGQo5JG4RmBoYZsk1HeoDrTxjrsZbLdFH34wCgTNuS8qrL0pUtjNABkwhtp7XLC6eMGpsrZo-L3GmLvsyUu_Vhon3crlftuzZsH8WuXQUxjnmKsiK/s715/images+%25286%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="429" data-original-width="715" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh4k8C1BRsrTaK9JUros4RTJ9V18w-HGQo5JG4RmBoYZsk1HeoDrTxjrsZbLdFH34wCgTNuS8qrL0pUtjNABkwhtp7XLC6eMGpsrZo-L3GmLvsyUu_Vhon3crlftuzZsH8WuXQUxjnmKsiK/s320/images+%25286%2529.jpeg" width="320" /></a></div><br /><div class="separator" style="clear: both; text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEinP9ZRFit8IKmIxBylMLbh2tVyT2HdZYmIfwOZZ2nXtqxbrQ2SJE4F-hTHTjrtG1koSwh2hMrfn1OtrjPJ0urUplIbuGjY3BrX1mCouZfw1JS8fwSbEK68_1V_y2mJitNHqCAvJg05liNb/s640/images+%25287%2529.jpeg" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" data-original-height="480" data-original-width="640" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEinP9ZRFit8IKmIxBylMLbh2tVyT2HdZYmIfwOZZ2nXtqxbrQ2SJE4F-hTHTjrtG1koSwh2hMrfn1OtrjPJ0urUplIbuGjY3BrX1mCouZfw1JS8fwSbEK68_1V_y2mJitNHqCAvJg05liNb/s320/images+%25287%2529.jpeg" width="320" /></a></div><br /><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;"><br /></span><p></p><p>The woman had seen lions and monkeys in her house. She was becoming disoriented and aggressive towards others, and was convinced that her husband was an impostor. She was in her mid-50s — decades older than the age at which psychosis typically develops — and had no psychiatric history. What she did have, however, was COVID-19. Hers was one of the first known cases of someone developing psychosis after contracting the disease</p><p>In the early months of the COVID-19 pandemic, doctors struggled to keep patients breathing, and focused mainly on treating damage to the lungs and circulatory system. But even then, evidence for neurological effects was accumulating. Some people hospitalized with COVID-19 were experiencing delirium: they were confused, disorientated and agitated. In April, a group in Japan published the first report of someone with COVID-19 who had swelling and inflammation in brain tissues. Another report described a patient with deterioration of myelin, a fatty coating that protects neurons and is irreversibly damaged in neurodegenerative diseases such as multiple sclerosis.</p><p>“The neurological symptoms are only becoming more and more scary,” says Alysson Muotri, a neuroscientist at the University of California, San Diego, in La Jolla.</p><p>The list now includes stroke, brain haemorrhage and memory loss. It is not unheard of for serious diseases to cause such effects, but the scale of the COVID-19 pandemic means that thousands or even tens of thousands of people could already have these symptoms, and some might be facing lifelong problems as a result.</p><p>Yet researchers are struggling to answer key questions — including basic ones, such as how many people have these conditions, and who is at risk. Most importantly, they want to know why these particular symptoms are showing up.</p><p>Although viruses can invade and infect the brain, it is not clear whether SARS-CoV-2 does so to a significant extent. The neurological symptoms might instead be a result of overstimulation of the immune system. It is crucial to find out, because these two scenarios require entirely different treatments. “That’s why the disease mechanisms are so important,” says Benedict Michael, a neurologist at the University of Liverpool, UK.</p><p><br /></p><p><span style="font-size: large;"><b>Affected brains.</b></span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">As the pandemic ramped up, Michael and his colleagues were among many scientists who began compiling case reports of neurological complications linked to COVID-19.</span></p><p><span style="background-color: white; color: #222222; font-family: Harding, Palatino, serif; font-size: 17px;">In a June paper5, he and his team analysed clinical details for 125 people in the United Kingdom with COVID-19 who had neurological or psychiatric effects. Of these, 62% had experienced damage to the brain’s blood supply, such as strokes and haemorrhages, and 31% had altered mental states, such as confusion or prolonged unconsciousness — sometimes accompanied by encephalitis, the swelling of brain tissue. Ten people who had altered mental states developed psychosis</span></p><p>Not all people with neurological symptoms have been seriously ill in intensive-care units, either. “We’ve seen this group of younger people without conventional risk factors who are having strokes, and patients having acute changes in mental status that are not otherwise explained,” says Michael.</p><p>A similar study1 published in July compiled detailed case reports of 43 people with neurological complications from COVID-19. Some patterns are becoming clear, says Michael Zandi, a neurologist at University College London and a lead author on the study. The most common neurological effects are stroke and encephalitis. The latter can escalate to a severe form called acute disseminated encephalomyelitis, in which both the brain and spinal cord become inflamed and neurons lose their myelin coatings — leading to symptoms resembling those of multiple sclerosis. Some of the worst-affected patients had only mild respiratory symptoms. “This was the brain being hit as their main disease,” says Zandi.</p><p>Less common complications include peripheral nerve damage, typical of Guillain–Barré syndrome, and what Zandi calls “a hodgepodge of things”, such as anxiety and post-traumatic stress disorder. Similar symptoms have been seen in outbreaks of severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS), also caused by coronaviruses. But fewer people were infected in those outbreaks, so less data are available.</p><p><br /></p><p>How many people?</p><p>But a major problem in quantifying cases is that clinical studies have typically focused on people with COVID-19 who were hospitalized, often those who required intensive care. The prevalence of neurological symptoms in this group could be “more than 50%”, says neurobiologist Fernanda De Felice at the Federal University of Rio de Janeiro in Brazil. But there is much less information about those who had mild illness or no respiratory symptoms.</p><p>That scarcity of data means it is difficult to work out why some people have neurological symptoms and others do not. It is also unclear whether the effects will linger: COVID-19 can have other health impacts that last for months, and different coronaviruses have left some people with symptoms for years</p><p>Infection or inflammation?</p><p>The most pressing question for many neuroscientists, however, is why the brain is affected at all. Although the pattern of disorders is fairly consistent, the underlying mechanisms are not yet clear, says De Felice.</p><p>Finding an answer will help clinicians to choose the right treatments. “If this is direct viral infection of the central nervous system, these are the patients we should be targeting for remdesivir or another antiviral,” says Michael. “Whereas if the virus is not in the central nervous system, maybe the virus is clear of the body, then we need to treat with anti-inflammatory therapies.”</p><p>Getting it wrong would be harmful. “It’s pointless giving the antivirals to someone if the virus is gone, and it’s risky giving anti-inflammatories to someone who’s got a virus in their brain,” says Michael.</p><p>There is clear evidence that SARS-CoV-2 can infect neurons. Muotri’s team specializes in building ‘organoids’ — miniaturized clumps of brain tissue, made by coaxing human pluripotent stem cells to differentiate into neurons.</p><p>In a May preprint, the team showed that SARS-CoV-2 could infect neurons in these organoids, killing some and reducing the formation of synapses between them. Work by immunologist Akiko Iwasaki and her colleagues at Yale University School of Medicine in New Haven, Connecticut, seems to confirm this using human organoids, mouse brains and some post-mortem examinations, according to a preprint published on 8 September. But questions remain over how the virus might reach people’s brains.</p><p>Because loss of smell is a common symptom, neurologists wondered whether the olfactory nerve might provide a route of entry. “Everyone was concerned that this was a possibility,” says Michael. But the evidence points against it.</p><p>A team led by Mary Fowkes, a pathologist at the Icahn School of Medicine at Mount Sinai in New York City, posted a preprint in late May describing post mortems in 67 people who had died of COVID-19. “We have seen the virus in the brain itself,” says Fowkes: electron microscopes revealed its presence. But virus levels were low and were not consistently detectable. Furthermore, if the virus was invading through the olfactory nerve, the associated brain region should be the first to be affected. “We’re simply not seeing the virus involved in the olfactory bulb,” says Fowkes. Rather, she says, infections in the brain are small and tend to cluster around blood vessels.</p><p>Michael agrees that the virus is hard to find in the brain, compared with other organs. Tests using the polymerase chain reaction (PCR) often do not detect it there, despite their high sensitivity, and several studies have failed to find any virus particles in the cerebrospinal fluid that surrounds the brain and spinal cord. One reason might be that the ACE2 receptor, a protein on human cells that the virus uses to gain entry, is not expressed much in brain cells.</p><p>“It seems to be incredibly rare that you get viral central nervous system infection,” Michael says. That means many of the problems clinicians are seeing are probably a result of the body’s immune system fighting the virus.</p><p>Still, this might not be true in all cases, which means that researchers will need to identify biomarkers that can reliably distinguish between a viral brain infection and immune activity. That, for now, means more clinical research, post mortems and physiological studies.</p><p>De Felice says that she and her colleagues are planning to follow patients who have recovered after intensive care, and create a biobank of samples including cerebrospinal fluid. Zandi says that similar studies are beginning at University College London. Researchers will no doubt be sorting through such samples for years. Although the questions they’re addressing have come up during nearly every disease outbreak, COVID-19 presents new challenges and opportunities, says Michael. “What we haven’t had since 1918 is a pandemic on this scale.”</p>Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com3tag:blogger.com,1999:blog-204734490966806252.post-20543581848714509262020-07-25T20:29:00.001-07:002020-07-25T20:29:23.042-07:00Study identifies six different "types" of COVID-19<div class="separator" style="clear: both; text-align: center;">
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<span style="background-color: white;">Study identifies six different "types" of COVID-19</span></h2>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">A new study of </span><span class="link" style="background-color: white; box-sizing: border-box; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;"><a data-ftag-appended="" data-invalid-url-rewritten-http="" href="https://www.cbsnews.com/feature/coronavirus/?ftag=CNM-00-10aab4i" style="box-sizing: border-box; color: #101010;">COVID-19</a></span><span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">, based on data from a symptom tracker app, determined that there are six distinct "types" of the disease involving different clusters of symptoms. The discovery could potentially open new possibilities for how doctors can better treat individual patients and predict what level of hospital care they would need.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">Researchers from King's College London studied data from approximately 1,600 U.K. and U.S. patients who regularly logged their symptoms in the COVID Symptom Tracker App in March and April.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">Typically, doctors will look for key symptoms such as cough, fever and loss of the sense of smell to detect COVID-19. The study, which has not been peer-reviewed, says the six different "types" of COVID-19 can vary by severity and come with their own set of symptoms.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">"I think it's very, very interesting," Dr. Bob Lahita, who is not affiliated with the study, told CBSN anchors Vladimir Duthiers and Anne-Marie Green. "Among the patients I see, those who recovered, many of them present different ways: some people with fever and some without fever, and some with nausea and vomiting, some people with diarrhea, etc."</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">The six clusters of symptoms outlined in the study are:</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;"><b>1)Flu-like with no fever</b>: </span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">Headache, loss of smell, muscle pains, cough, sore throat, chest pain, no fever.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;"><b>2)Flu-like with fever: </b></span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">Headache, loss of smell, cough, sore throat, hoarseness, fever, loss of appetite</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;"><b>3)Gastrointestinal:</b></span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;"> Headache, loss of smell, loss of appetite, diarrhea, sore throat, chest pain, no cough.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;"><b>4)Severe level one, fatigue: </b></span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">Headache, loss of smell, cough, fever, hoarseness, chest pain, fatigue.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;"><b>5)Severe level two, confusion:</b></span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;"> Headache, loss of smell, loss of appetite, cough, fever, hoarseness, sore throat, chest pain, fatigue, confusion, muscle pain.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;"><b>6)Severe level three, abdominal and respiratory: </b></span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">Headache, loss of smell, loss of appetite, cough, fever, hoarseness, sore throat, chest pain, fatigue, confusion, muscle pain, shortness of breath, diarrhea, abdominal pain.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">The first level, "flu-like with no fever," is associated with headaches, loss of smell, muscle pains, cough, sore throat and chest pain. Patients at this level have a 1.5% chance of needing breathing support such as oxygen or a ventilator.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">The second type, "flu-like with fever," includes symptoms like loss of appetite, headache, loss of smell, cough, sore throat, hoarseness and fever. Researchers say about 4.4% of patients at this level needed breathing support.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">Patients with the third type, simply described as "gastrointestinal," do not have a cough as part of their illness. Instead, they experience headache, diarrhea, loss of smell, loss of appetite, sore throat and chest pain, and about 3.3% needed breathing support.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">In type four, or "severe level one," patients experience fatigue along with headache, loss of smell, cough, fever, hoarseness and chest pain. Patients at this level needed breathing support at a rate of 8.6%</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">Type five, "severe level two," includes the symptoms of type four along with loss of appetite, sore throat and muscle pain, and is mainly distinguished by confusion.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">"That means you don't know where you are or where you live, whether you are in or out of the hospital, who your relatives are," Lahita explained. "That is very scary." Almost 10% of patients at that level need breathing support.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">"That means you don't know where you are or where you live, whether you are in or out of the hospital, who your relatives are," Lahita explained. "That is very scary." Almost 10% of patients at that level need breathing support.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">The most severe type of COVID-19 is referred to as "severe level three, abdominal and respiratory," and has all the above symptoms along with abdominal pain, shortness of breath and diarrhea. Nearly 20% of these patients need breathing support.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">"Those are the severe level threes who wind up on a ventilator, and then it is touch-and-go as to whether they survive the infection entirely," Lahita said.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">The U.K. researchers also found that only 16% of patients with type one COVID-19 required hospitalization, compared with nearly half of the patients with type six.</span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">Patients in the severe clusters also tended to be older or with pre-exisiting conditions and weakened immune systems, compared to those in the first three. </span></div>
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<span style="background-color: white; color: #101010; font-family: "publico text" , serif; font-size: 19.04px;">Scientists hope the discovery, once further studied, could help predict what types of care patients with COVID-19 might need, and give doctors the ability to predict which patients would fall into which category. </span></div>
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Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com6tag:blogger.com,1999:blog-204734490966806252.post-85244751135770338272020-06-21T22:13:00.004-07:002020-06-21T22:18:16.270-07:003 COVID-19 treatment drugs available in India today<div class="separator" style="clear: both; text-align: center;">
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEj22jCw0QphvJxUgnOxBmgJ0Nkc4dVibsurwDMiiMl32ul7Z5XUesnrPCL2EN7bzwxhZYuGNnEXIJU86u1xfmE1NF3-Pava8JrRMrdBuBCUII_ivjBeOL5B4ZKLnFg8AMLIHu3X46i_DPgV/s1600/Injections-Vs.-Oral-Medications_-Which-Is-More-Effective_.Featured-1080x630.jpg" imageanchor="1" style="clear: left; float: left; margin-bottom: 1em; margin-right: 1em;"><img border="0" data-original-height="630" data-original-width="1080" height="232" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEj22jCw0QphvJxUgnOxBmgJ0Nkc4dVibsurwDMiiMl32ul7Z5XUesnrPCL2EN7bzwxhZYuGNnEXIJU86u1xfmE1NF3-Pava8JrRMrdBuBCUII_ivjBeOL5B4ZKLnFg8AMLIHu3X46i_DPgV/s400/Injections-Vs.-Oral-Medications_-Which-Is-More-Effective_.Featured-1080x630.jpg" width="400" /></a></div>
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<span style="font-size: small;">Coronavirus pandemic has killed over 4.7 lakh people across the globe. In India, the death toll is over 13,000. The good news is that DGCI has given nod to 3 companies to roll out their COVID-19 treatment medicine in India. These companies are -- Cipla, Glenmark and Hetero. And the three medicines are called Cipremi, FabiFlu and Covifor.</span></h2>
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<span style="color: #010101; font-family: "roboto" , sans-serif;">The three have reportedly shown good results so far and will be now slowly put into use under strict medical observat</span><span style="color: #010101; font-family: "roboto" , sans-serif;">ion.</span></div>
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<strong style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; margin: 0px; outline: 0px; padding: 0px; vertical-align: baseline;">Here is everything we know about these drugs.</strong></h2>
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<span style="background-position: 0px center; border: 0px; margin: 0px; outline: 0px; padding: 0px; vertical-align: baseline;"><strong style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; margin: 0px; outline: 0px; padding: 0px; vertical-align: baseline;">CIPREMI</strong></span></div>
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Cipla has launched its own remedesivir under the name of Cipremi. The medicine is lyophilized powder for injection 100mg. </div>
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The drug will be marketed by both the government and market channels. </div>
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The drug has been approved for adult and paediartric patients hospitalised due to COVID-19 infection. The drug is most affective on those who need oxygen support.</div>
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The drug is most affective on those who need oxygen support. </div>
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Cipla is yet to disclose the pricing for the drug.</div>
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<strong style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-family: Roboto, sans-serif; font-size: 16px; margin: 0px; outline: 0px; padding: 0px; vertical-align: baseline;"><br /></strong></div>
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<strong style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-family: Roboto, sans-serif; font-size: 16px; margin: 0px; outline: 0px; padding: 0px; vertical-align: baseline;">FABIFLU</strong></div>
<ul style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 26px; margin: 0px; outline: 0px; padding: 10px 0px 0px 30px; vertical-align: baseline;">
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-size: 16px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">Priced at Rs 3,500 for 34 tablets, the dosage is 200 mg X 9 tablets on day one and 200 mg X 4 tablets a day for 14 days.</li>
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-size: 16px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">Global trials show the efficacy of over 80-88%; Japan, Bangladesh and UAE already use the drug for COVID-19 treatment.</li>
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-size: 16px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">The drug will be available both through hospitals and the retail channel.</li>
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-size: 16px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">Reportedly, Strides Pharma, Brinton Pharmaceuticals, Lasa Supergenerics and Optimus Pharma among firms readying its launch</li>
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-size: 16px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">Glenmark had developed the active pharmaceutical ingredient (API) and the formulation for FabiFlu through in-house R&D.</li>
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-size: 16px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">Favipiravir is backed by strong clinical evidence, showing encouraging results in patients with mild to moderate Covid-19.</li>
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-size: 16px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">Patients from over 10 leading government and private hospitals were enrolled for the study.</li>
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; font-size: 16px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">It offers rapid reduction in viral load within four days and provides faster symptomatic and radiological improvement. </li>
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<span style="font-family: "roboto" , sans-serif;"><span style="font-family: "Times New Roman"; font-size: 16px;"> </span><strong style="background-position: 0px center; border: 0px; font-size: 16px; margin: 0px; outline: 0px; padding: 0px; vertical-align: baseline;">COVIFOR</strong></span></div>
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<ul style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; color: #010101; font-family: Roboto, sans-serif; font-size: 16px; font-stretch: normal; font-variant-east-asian: normal; font-variant-numeric: normal; line-height: 26px; margin: 0px; outline: 0px; padding: 10px 0px 0px 30px; vertical-align: baseline;">
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">The drug will be available in 100 mg injectable form which has to be administered intravenously in a hospital setting under the supervision of a healthcare professional. It is not a drug you can take at home. </li>
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">The company is sure about enough stock to cater to the present needs of the medicine. </li>
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">Hetero has confirmed that Covifor would cost between 5,000 to 6,000 per dose. </li>
<li style="background-attachment: initial; background-clip: initial; background-image: initial; background-origin: initial; background-position: 0px center; background-repeat: initial; background-size: initial; border: 0px; line-height: 26px; margin: 0px; outline: 0px; padding: 0px 0px 10px; vertical-align: baseline;">The COVID-19 treatment by Covifor will cost not more than 30,000 per patient. Six dozes of the medicine will be given in this timeframe. </li>
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Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-1573953511529458142020-06-15T09:19:00.001-07:002020-06-15T09:22:32.714-07:00FSH Followed by HMG vs FSH Plus HMG in IVF<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiP60JKHeE-Ixr9SePy0iXJlbWZNJ3v05DaGAUXewy0u8DeMUd2qdHd2wLcsw8I6MhDaZaLCvoFqgBGLDaYJkcj8A0V6150-Pj7JpausMS0GbdMCOGoG-joKeyD9lSNUK0GXPLIqpEIgt50/s1600/F2.large.jpg" imageanchor="1"><img border="0" data-original-height="866" data-original-width="1280" height="217" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiP60JKHeE-Ixr9SePy0iXJlbWZNJ3v05DaGAUXewy0u8DeMUd2qdHd2wLcsw8I6MhDaZaLCvoFqgBGLDaYJkcj8A0V6150-Pj7JpausMS0GbdMCOGoG-joKeyD9lSNUK0GXPLIqpEIgt50/s320/F2.large.jpg" width="320" /></a><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjOLZ0f_QTVnKUh_aRHNItDqZ5AYg7wdj9QwqwoEa5LUgiiraOtTK-_lWNVZMiOX4MC1TJcEUI6rW7dub-WhXAsTQLfNmFHihBUSS9OLjyvMrGRh4jzlJZNUrkek1MCmxF_TQPL3pCYuE9z/s1600/GettyImages-107702265-56c0358a3df78c0b138e8365.jpg" imageanchor="1"><img border="0" data-original-height="1200" data-original-width="1600" height="240" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjOLZ0f_QTVnKUh_aRHNItDqZ5AYg7wdj9QwqwoEa5LUgiiraOtTK-_lWNVZMiOX4MC1TJcEUI6rW7dub-WhXAsTQLfNmFHihBUSS9OLjyvMrGRh4jzlJZNUrkek1MCmxF_TQPL3pCYuE9z/s320/GettyImages-107702265-56c0358a3df78c0b138e8365.jpg" width="320" /></a><br />
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<br /> FSH Followed by HMG vs FSH Plus HMG in IVF </h2>
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<b>Brief description of study </b></h2>
The aim of this study is to compare the clinical outcomes of sequential administration of FSH and HP-hMG FSH alone versus concomitant administration of FSH and HP-hMG during controlled ovarian stimulation in IVF cycles.<br />
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<br /><b>Detailed Study Description</b></h2>
Women who are planned to be subjected to IVF/ICSI through COS by long GnRHa protocol will be assessed for possibility of participation in our study. Eligible participants in our study will be those with regular menstrual cycle (21-35 days) and normal uterine anatomy (confirmed by transvaginal ultrasound examination and in some cases hysteronsalpingography and hysteroscopy).<br />
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Women with any of the following criteria will be excluded from the study: 1) age < 20 or > 37 years; 2) body mass index (BMI) < 18 or > 25 kg/m2; 3) low ovarian reserve (AFC < 7 and/or AMH < 1.1 ng/ml); 4) presence of polycystic ovarian syndrome (PCOS), endometrioma or hydrosalpinx; 5) history of chemotherapy, radiotherapy or ovarian surgery; 6) the husband needs testicular biopsy to obtain sperm; or 7) previous implantation failure.<br />
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A written informed consent will be taken from each women selected to participate before inclusion in the study. All women participating in the study will start GnRHa on day 21 of the preceding cycle and when down regulation occurs each woman will be randomly allocated into one of the two groups; group 1 and group 2. Women in group 1 will receive 225 IU FSH alone from day one of ovarian stimulation and when the follicular diameters reaches 10-12 mm, the 150 IU HP-hMG will substitute FSH and continued to the day of triggering. Women in group 2 will receive 150 IU FSH plus 75 IU HP-hMG from day one of ovarian stimulation and 150IU HP-HMG when the follicular diameters reaches 10-12 mm till day of triggering. The randomization will be simple and balanced (1:1) and will be carried out by a nurse through sealed, unlabeled, opaque envelopes containing computer-generated random numbers. The data assesor will be blinded to group assignment.<br />
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In both groups, estradiol and LH will be measured on the third day of menstruation before start of stimulation and on day 6 of stimulation TVS will be performed. Progesterone and E2 will be measured and on day of triggering. The primary outcome measure of this study will be the ongoing pregnancy rate. The secondary outcomes measures will be cancellation rate, the number of oocytes retrieved, the number of embryos, the number of vitrified embryos, the clinical pregnancy rate, the implantation rate, OHSS rate, multiple pregnancy rate, and the miscarriage rate.Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-35297186166700904432020-06-13T09:14:00.000-07:002020-06-13T09:16:04.942-07:00Cost-effectiveness analysis of GnRH-agonist long-protocol and GnRH-antagonist protocol for in vitro fertilization<div class="c-article-header" style="background-color: white; color: #222222; font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 18px; margin: 0px 0px 40px; padding: 0px;">
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Cost-effectiveness analysis of GnRH-agonist long-protocol and GnRH-antagonist protocol for <i>in vitro</i> fertilization</h1>
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<li class="c-author-list__item" itemprop="author" itemscope="itemscope" itemtype="http://schema.org/Person" style="display: inline; margin-left: 0px; padding-right: 0px;"><span itemprop="name">Miaomiao Jing</span>, </li>
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<li class="c-author-list__item js-smaller-author-etal" itemprop="author" itemscope="itemscope" itemtype="http://schema.org/Person" style="display: inline; margin-left: 0px; padding-right: 0px;"><span itemprop="name">Qi Chen</span>, </li>
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<li class="c-author-list__item" itemprop="author" itemscope="itemscope" itemtype="http://schema.org/Person" style="display: inline; margin-left: 0px; padding-right: 0px;"><span itemprop="name">Runju Zhang<svg class="u-icon" height="16" width="16"><use xlink:href="#global-icon-email" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></span> </li>
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, Article number: <span data-test="article-number">8732</span> (<span data-test="article-publication-year">2020</span>) <span id="goog_290097558"></span><span id="goog_290097559"></span></div>
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Abstract</h2>
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The gonadotropin releasing hormone agonist (GnRH-a) long-protocols and the GnRH-antagonist protocols are two commonly used protocols for <i>in vitro</i> fertilization (IVF), but their cost-effectiveness has not been studied, especially in China. A retrospective study involving 1638 individuals in GnRH-a long-protocol and 621 in GnRH-antagonist protocol were conducted and a decision tree model analysis was used to analyze the cost-effectiveness. Both direct and indirect costs were calculated. As a result, during the fresh embryo transplantation cycles, there was no significant difference in the rate of ongoing pregnancy between the two protocols, the average cost of per ongoing pregnancy in the GnRH-antagonist protocol was $ 16970.85, and that in the GnRH-agonist long-protocol was $19902.24. The probability of cumulative ongoing pregnancy per start cycle was estimated at 60.65% for the GnRH-antagonist protocol and 71.6% for the GnRH-agonist long-protocol (P < 0.01). Considering the cumulative ongoing pregnancy rate, the mean costs per ongoing pregnancy were estimated at $8176.76 and at $7595.28 with GnRH-antagonist protocol and GnRH-agonist long protocol, respectively. In conclusion, in fresh embryo transplantation cycle, the GnRH-antagonist protocol has economic advantage. However, the GnRH-agonist long protocol is more cost effective considering the cumulative ongoing pregnancy rate in the fresh embryo and frozen embryo transplantation cycles.</div>
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<h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec1" style="border-bottom: 2px solid rgb(213, 213, 213); font-size: 2.4rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px 0px 8px;">
Introduction</h2>
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According to a World Health Organization (WHO) report, about 48.5 million couples are affected by infertility worldwide in 2010. With the refinement and development of assisted reproductive technology (ART), increasing number of infertile couples seek ART. The European Society of Human Reproduction and Embryology (ESHRE) reported that the world-wide number of babies born as a result of ART has reached an estimated total of 8 million since the world’s first, Louise Brown, was born in July 1978. Up to now, ART is the most important method to treat infertility in the world.</div>
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Study has shown that cumulative live birth rates are increased with the number of oocytes obtained. Therefore, controlled ovarian hyperstimulation (COH) is an important process to obtain a set number of oocytes for IVF. The GnRH-agonists were introduced into IVF in the late 1980s and the GnRH-agonist long-protocol is still the most frequently used protocol in most centers worldwide<span style="font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 4" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-65558-0#ref-CR4" id="ref-link-section-d141997e451" style="background-color: transparent; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Coccia, M. E., Comparetto, C., Bracco, G. L. & Scarselli, G. GnRH antagonists. European journal of obstetrics, gynecology, and reproductive biology 115(Suppl 1), S44–56,
https://doi.org/10.1016/j.ejogrb.2004.01.033
(2004).">4</a></span>. The basic principle is to use gonadotropin-releasing hormone agonist (GnRH-a) to regulate pituitary and stimulate follicular growth with exogenous gonadotropin hormone, and avoid endogenous luteinizing hormone (LH) surge before oocyte retrieval. Since 1990s, GnRH antagonists were used in COH, this protocol competitively blocks pituitary GnRH receptors, inducing a rapid, reversible suppression of gonadotrophin secretion and preventing and interrupting LH surges. The GnRH-antagonist protocol have been widely adopted in IVF due to these advantages.</div>
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IVF is a protracted and costly process. In most developed countries, IVF is covered by insurance or subsidized. This is not the case in developing countries. In China, the cost of IVF is on patients. High costs discourage low-income infertility couples from seeking ART treatment.</div>
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While both protocols are commonly used today, little is known about their cost-effectiveness, especially in China. Studies have shown that hormonal stimulation covered the main part of the costs per cycle. Moolenaar <i>et al</i>. reported that most economic studies about ART were performed in countries from mainland Europe (38%) and the United States (34%). In China, there has been few economic researches on COH protocols. Herein, we performed a retrospective study on comparing the cost-effectiveness of the two protocols using a single center data in China. Choosing a cost-effective protocol, can not only ease the financial pressure on couples, but also provide reference for medical decision-making.</div>
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<h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec2" style="border-bottom: 2px solid rgb(213, 213, 213); font-size: 2.4rem; line-height: 1.24; margin: 0px 0px 8px; padding: 0px 0px 8px;">
Methods</h2>
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Patients</h3>
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Individuals who came to the Reproductive Center of Women’s Hospital, Zhejiang University School of Medicine for their first cycle of IVF treatment from 1 January 2015 to 31 December 2017 were included. Inclusion criteria were as follows: 20 < age ≤ 38 years, regular ovulatory cycles every 21–35 days, total antral follicle count (AFC) ≥ 5, first cycle of IVF treatment, COH planned using the GnRH-agonist long-protocol or the GnRH-antagonist protocol, IVF fertilization. Exclusion criteria were the use of donor oocytes or frozen-thawed oocytes for fertilization, other protocols for COH, ICSI fertilization. Patient demographics are presented in Table 1.</div>
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Table 1 Patient demographics and infertility treatment-related characteristics.</h1>
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From: Cost-effectiveness analysis of GnRH-agonist long-protocol and GnRH-antagonist protocol for <i>in vitro</i> fertilization</div>
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<table class="data last-table" style="border-collapse: collapse; border-spacing: 0px; border: 2px solid rgb(213, 213, 213); font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1.6rem; margin-bottom: 0px; width: 998px;"><thead class="c-article-table-head" style="border-bottom: 5px solid rgb(149, 149, 149);">
<tr style="border-bottom: 1px solid rgb(213, 213, 213);"><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Characteristics</th><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">GnRH-agonist long-protocol</th><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">GnRH-antagonist protocol</th><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">P value</th></tr>
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<tr style="border-top: 0px;"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Age(year)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">29.26 ± 3.36</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">29.54 ± 3.32</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.078</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Duration of infertility (year)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">3.12 ± 2.34</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">3.28 ± 2.51</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.148</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Height (cm)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">159.57 ± 6.02</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">159.56 ± 4.60</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.964</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Weight (kg)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">55.52 ± 7.57</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">55.89 ± 7.26</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.305</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">BMI (kg/m<span style="font-size: 12px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">2</span>)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">21.76 ± 2.81</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">21.95 ± 2.69</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.144</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">AFC (n)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">14.52 ± 4.06</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">14.38 ± 4.30</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.463</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">bFSH (IU/L)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">6.22 ± 1.67</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">6.73 ± 1.79</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><0.01</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">bLH (IU/L)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">5.67 ± 3.18</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">5.58 ± 2.99</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.52</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">bE2 (pmol/l)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">116.67 ± 67.72</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">115.40 ± 64.83</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.688</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Duration of Gonadotropin stimulation (day)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">10.82 ± 1.78</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">9.53 ± 1.87</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><0.01</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Gonadotropin used (IU)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">2021.09 ± 668.56</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1823.78 ± 561.22</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><0.01</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Number of oocytes obtained (n)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">13.90 ± 6.58</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">11.89 ± 6.70</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><0.01</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Number of embryos available (n)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">4.42 ± 3.43</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">3.98 ± 3.27</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.005</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Number of frozen embryos (n)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">3.72 ± 3.74</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">3.20 ± 3.57</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.005</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Average number of embryos transferred (n)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1.810 ± 0.40</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1.79 ± 0.42</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.349</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Number of Frozen embryo transplantable cycle (n)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">2.05</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1.78</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">—</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Clinical pregnancy rate in the fresh embryo transplantation cycle</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">47.26%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">49.44%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.487</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Ongoing pregnancy rate of fresh embryo transplantation</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">38.77%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">37.22%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.613</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Cost in fresh embryo transplantation($)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">19902.24</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">16970.85</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">—</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Cumulative ongoing pregnancy rate</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">71.6%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">60.65%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><0.01</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Cumulative multiple pregnancy rate</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">17.6%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">18.4%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.667</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">OHSS rate</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">4.9%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">2.0%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.001</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Cumulative ectopic pregnancy rate</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">2.9%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">4.0%</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.26</td></tr>
</tbody></table>
</div>
</div>
<div class="c-article-table-footer" style="margin: 0px; padding: 10px;">
<ol style="list-style: none; margin: 0px; padding: 0px;">
<li>AFC: antral follicle count; bFSH: basal follicle stimulating hormone; bLH: basal luteinizing hormone; bE2: basal estrogen; OHSS: ovarian hyperstimulation syndrome.</li>
</ol>
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<div style="margin-bottom: 28px; overflow-wrap: break-word; padding: 0px;">
<span style="background-color: initial; font-size: 2.4rem;">COH protocols</span></div>
<div style="margin-bottom: 28px; overflow-wrap: break-word; padding: 0px;">
GnRH-agonist long-protocol: A short-acting GnRH-a (Triptorelin, Ferring AG, Germany) was administrated daily in the mid luteal phase of the preceding cycle. 14 days later, follicular ultrasonography, serum LH, FSH and E2 were examined and 150–300IU recombination follicle stimulating hormone (r-FSH, Gonal-F, Merck Serono, Switzerland) was initiated daily when FSH and LH < 5 IU/L and E2 < 50 pg/ml. GnRH-a was continued until trigger. During the stimulation, according to ovarian response evaluated by transvaginal ultrasonography and serum hormone level, dose of r-FSH was adjusted as needed, human menopausal gonadotropin (HMG) or recombinant luteinizing hormone(r-LH) or growth hormone (GH) was added as needed.</div>
<div style="margin-bottom: 28px; overflow-wrap: break-word; padding: 0px;">
GnRH-antagonist protocol: 150–300IU r-FSH was initiated on day 2 or 3 of the menstrual cycle until trigger. The dosage of r-FSH was adjusted and HMG or r-LH or GH was added according to the ovarian response evaluated by transvaginal ultrasonography and serum hormone level. 0.25 mg GnRH-A (Cetrorelix; Merck Serono, France) was used daily until trigger when the leading follicles reached a mean diameter of 14 mm.</div>
<div style="margin-bottom: 28px; overflow-wrap: break-word; padding: 0px;">
For both protocols, if three follicles reached a mean diameter of 17 mm or two follicles reached a mean diameter of 18 mm, r-HCG (Ovidrel, Serono, Italy) was administered subcutaneously. Oocyte retrieval was performed 36 h after HCG injection by transvaginal ultrasound-guided single-lumen needle aspiration. Luteal phase support was initiated on day 1 after oocyte retrieval. Fresh embryo transplantation was carried out 72 h after oocyte retrieval. Fresh cycles were canceled if patients had endometrial thickness <7 mm, high risk of Ovarian hyperstimulation syndrome (OHSS) (E2 ≥ 5000 pg/ml on the trigger day, the number of oocytes obtained ≥20), no available embryos or other personal reasons. For those patients without fresh embryo transplantation or without ongoing pregnancy after fresh embryo transplantation, if excess frozen embryos were available and pregnancy test was negative, the frozen embryo transplantation (FET) was performed until no embryos remained or ongoing pregnancy was achieved. Clinical pregnancy was defined as a gestational sac observed by vaginal ultrasound. Ongoing pregnancy was defined as a pregnancy continuing 12 weeks without miscarriage.</div>
<h3 class="c-article__sub-heading" id="Sec5" style="font-size: 2.4rem; font-weight: 400; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">
Structure of the model</h3>
<div style="margin-bottom: 28px; overflow-wrap: break-word; padding: 0px;">
We constructed a decision tree model to analyze the cost-effectiveness of the GnRH-agonist long-protocol and GnRH-antagonist protocol in the fresh embryo transplantation and frozen embryo transplantation cycle (Fig. 1 and Fig. 2). Each route in the diagram represents possible steps in IVF. Each intersection is followed by a possible situation. Since the cumulative ongoing pregnancy rate was calculated by following utilization of all fresh and frozen embryos after the first IVF cycle, the number of transplants was based on available embryos. The terminal nodes of the model were: “No oocytes”, “No embryos”, “Ongoing pregnancy”, “No ongoing pregnancy”. Since each patient may have different situations after entering the cycle, we made relevant assumptions for the model for the convenience of calculation as shown in Table 2.</div>
<div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-test="figure" id="figure-1" style="border: 5px solid rgb(213, 213, 213); clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;">
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<figure style="margin: 0px;"><figcaption><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig1" style="display: block; font-weight: bolder; margin-bottom: 8px;">Figure 1</span></figcaption><div class="c-article-section__figure-content" style="margin: 0px 0px 16px; padding: 0px;">
<div class="c-article-section__figure-item" style="display: inline-block; margin: 0px; max-width: 100%; padding: 0px;">
<picture><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-65558-0/MediaObjects/41598_2020_65558_Fig1_HTML.png?as=webp" type="image/webp"></source></picture></div>
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Decision tree model for fresh embryo transplantation. Figure 1 shows the process of fresh embryo transplantation. The number under each node in the figure is the correlation probability of this <span style="font-size: 18px; font-weight: bolder;">Figure 2</span></div>
<div style="overflow-wrap: break-word; padding: 0px;">
</div>
<div style="overflow-wrap: break-word; padding: 0px;">
<img alt="Figure 1" height="460" src="https://media.springernature.com/full/springer-static/image/art%3A10.1038%2Fs41598-020-65558-0/MediaObjects/41598_2020_65558_Fig1_HTML.png" width="640" /><span style="font-size: 1.6rem;">Decision tree model for frozen embryo transplantation. Figure 2 shows the frozen embryo transplantation process after fresh embryo transplantation. The number under each node in the figure is the correlation probability of this step.</span></div>
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<span style="font-size: 1.6rem;"><br /></span></div>
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<img alt="Figure 2" height="553" src="https://media.springernature.com/full/springer-static/image/art%3A10.1038%2Fs41598-020-65558-0/MediaObjects/41598_2020_65558_Fig2_HTML.png" width="640" /></div>
<div style="overflow-wrap: break-word; padding: 0px;">
<span style="font-size: 18px;">Decision tree model for frozen embryo transplantation. Figure 2 shows the frozen embryo transplantation process after fresh embryo transplantation. The number under each node in the figure is the correlation probability of this step.</span></div>
</div>
</div>
</figure></div>
<div class="c-article-section__figure js-c-reading-companion-figures-item" data-container-section="figure" data-test="figure" id="figure-2" style="border: 5px solid rgb(213, 213, 213); clear: both; margin: 0px 0px 24px; max-width: 100%; padding: 20px 10px;">
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<figure style="margin: 0px;"><div class="u-text-right u-hide-print" style="margin: 0px; padding: 0px; text-align: right;">
<span style="background-color: initial; font-weight: bolder;">Table 2 Assumptions in the model.</span></div>
</figure></div>
<div class="c-article-table" data-container-section="table" data-test="inline-table" id="table-2" style="border: 5px solid rgb(213, 213, 213); clear: both; margin: 0px 0px 24px; padding: 20px 10px;">
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<figure style="margin: 0px;"><figcaption class="c-article-table__figcaption" style="line-height: 1.4; margin-bottom: 16px;"><div class="c-article-table-container" style="margin: 0px 0px 24px; padding: 0px;">
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<table class="data last-table" style="border-collapse: collapse; border-spacing: 0px; border: 2px solid rgb(213, 213, 213); font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1.6rem; margin-bottom: 0px; width: 998px;"><thead class="c-article-table-head" style="border-bottom: 5px solid rgb(149, 149, 149);">
<tr style="border-bottom: 1px solid rgb(213, 213, 213);"><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Assumptions</th></tr>
</thead><tbody>
<tr style="border-top: 0px;"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">1.r-LH and GH were added according to the development of follicles, not all patients used it, so their costs were not included in the total cost.</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">2. In GnRH-agonist long-protocol, there were an average of 5 blood tests and follicular ultrasonography examinations, while in the GnRH-antagonist protocol, there were 4 blood tests and follicular ultrasonography examinations.</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">3. Artificial cycle drug was unified as estradiol valerate tablets, with an average of 4 follicular ultrasound tests. There were 4 follicular ultrasound tests on average in the natural cycle.</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">4. The corpus luteum support drugs were unified as estradiol valerate + dydrogesterone tablets + progesterone vaginal sustained release gel in fresh cycle and estradiol valerate + dydrogesterone tablets + progesterone capsules in frozen cycle.</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">5. The cost of lost work were calculated based on the per capita disposable income of the three-year residents in 2015–2017.</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">6. The transportation fee was calculated according to the second-class fare of the trains from various cities in Zhejiang Province to Hangzhou.</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">7. The bed fee during hospitalization was calculated at a rate of ¥40 per day for a three-person room.</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">8. The number of frozen embryo transfer cycles is calculated based on the number of frozen embryos and the number of transplanted embryos.</td></tr>
</tbody></table>
</div>
</div>
</div>
</figcaption><div class="u-text-right u-hide-print" style="margin: 0px; padding: 0px; text-align: right;">
<svg class="u-icon" height="16" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"></use></svg></div>
</figure></div>
<h3 class="c-article__sub-heading" id="Sec6" style="font-size: 2.4rem; font-weight: 400; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">
Transition probabilities</h3>
<div style="margin-bottom: 28px; overflow-wrap: break-word; padding: 0px;">
The transition probabilities for the various health states in this decision tree model were derived from the clinical data of included infertility patients. The transition probability of each step is shown in Fig. 1 and Fig. 2.</div>
<h3 class="c-article__sub-heading" id="Sec7" style="font-size: 2.4rem; font-weight: 400; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">
Cost analysis</h3>
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This study was performed from a patient’s perspective. Both direct and indirect costs were included in the analysis. Direct medical costs included drug, ultrasound, laboratory, surgery and care costs. Medications included those used in ovulation stimulation trigger and luteal support. The cost of the drug was equal to the unit cost of the drug multiplied by the total amount used. When calculating the cost of IVF, it is prudent to include the treatment costs for complications, primarily OHSS. Therefore, the cost of treatment for OHSS was also calculated in the total cost of the patient. In this study, transportation costs were included as direct non-medical expenses. These were calculated as the average fare from each city in Zhejiang province to Hangzhou. Indirect costs included the cost of lost work. The cost of lost work was calculated according to the per capita disposable income. Intangible economic burden generally refers to the decline in the quality of life or other costs caused by illness in this case, IVF, and also included other costs not reflected in the direct and indirect costs. Thus, considering the difficulty in calculating these intangible costs, such costs were not included in this study. Cycle costs were relatively stable during the study period, so discounting is not considered. The specific costs are shown in Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-65558-0#Tab3" style="background-color: transparent; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;">3</a>.</div>
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<span style="background-color: initial; font-weight: bolder;">Table 3 The cost involved in IVF.</span></div>
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<table class="data last-table" style="border-collapse: collapse; border-spacing: 0px; border: 2px solid rgb(213, 213, 213); font-family: -apple-system, BlinkMacSystemFont, "Segoe UI", Roboto, Oxygen-Sans, Ubuntu, Cantarell, "Helvetica Neue", sans-serif; font-size: 1.6rem; margin-bottom: 0px; width: 998px;"><thead class="c-article-table-head" style="border-bottom: 5px solid rgb(149, 149, 149);">
<tr style="border-bottom: 1px solid rgb(213, 213, 213);"><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Drugs</th><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Costs ($)</th><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Operation</th><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Costs ($)</th><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Non-medical expenses</th><th class="u-text-left " style="background: rgb(238, 238, 238); border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; text-align: left; vertical-align: top;">Costs ($)</th></tr>
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<tr style="border-top: 0px;"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Triptorelin(0.1 mg)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">16.33</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Oocyte retrieval</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">305.81</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Single transportation fee</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">11.61</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Gonal-F(450IU)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">230.26</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Anesthetic Fee</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">96.02</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">The daily cost of lost work</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">10.02</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">HMG(75IU)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">3.18</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">IVF</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">305.81</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Daily nursing expenses during hospitalization</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">3.67</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Cetrorelix(0.25 mg)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">52.72</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Embryo transplantation</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">244.65</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Bed fee per day during hospitalization</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">6.12</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Ovidrel(6500IU)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">29.5</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Embryo cryopreservation</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">611.62</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"></td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Estradiol valerate tablets(tablet)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.26</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Embryos thawing</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">76.45</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"></td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Dydrogesterone tablets(tablet)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.83</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Follicular ultrasonography</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">6.42</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"></td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Progesterone vaginal sustained release gel</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">11.47</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Serum FSH,LH,E2,Ptest</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">4.89</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"></td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">Progesterone capsules (capsule)</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.43</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><span style="font-weight: bolder;">Serum HCG</span></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">6.12</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"></td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;"><br /></td></tr>
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<h3 class="c-article__sub-heading" id="Sec8" style="font-size: 2.4rem; font-weight: 400; line-height: 1.24; margin: 0px 0px 8px; padding: 0px;">
Statistical analyses</h3>
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The statistical software package SPSS22 was used for data analysis. When the measurement data matched the homogeneity of variance, the independent sample t test was used; when the data didn’t match the homogeneity of variance, the Mann-Whitney test was used. The chi-square test or Fisher’s exact probability method were used for the comparison of the rates, and P < 0.05 was considered statistically significant. Cost-effectiveness analysis and incremental cost-effectiveness analysis were used to evaluate the costs and effects between the two protocols. Sensitivity analyses were then performed to assess the stability of the model.</div>
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Ethical approval</h3>
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The study was approved by the Ethics Committee of Women’s Hospital School of Medicine Zhejiang University. All patients meeting the inclusion criteria signed the informed consent. And this study complied with declaration of Helsinki/relevant guidelines for the study on humans.</div>
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Results</h2>
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About 2259 infertility patients met inclusion criteria (1638 in GnRH-agonist long-protocol and 621 in GnRH-antagonist protocol). There were no differences in patient ages, BMI, duration of infertility, cycle day 3 LH levels, cycle day 3 E2 levels and AFC. The duration of gonadotrophin stimulation and dose of gonadotropin used, number of oocytes obtained and number of available embryos in GnRH-agonist long-protocol were higher than that in GnRH-antagonist protocol. The clinical pregnancy rate and ongoing pregnancy rate for GnRH-agonist long-protocol and GnRH-antagonist protocol in the fresh embryo transplantation cycle were not different (47.26% vs. 49.44%, P = 0.487; 38.77% vs. 37.22%, P = 0.613). When all available embryos had been utilized, the rate of cumulative ongoing pregnancy per start cycle was 60.65% for GnRH-antagonist protocol and 71.6% for GnRH-agonist long-protocol (P < 0.01). The OHSS rate was lower in GnRH-antagonist protocol (2.0% vs 4.9%, P = 0.001). The cumulative multiple pregnancy (17.6% vs 18.4%, P = 0.667) rate and the cumulative ectopic pregnancy rate (2.9% vs 4.0%, P = 0.26) in GnRH-agonist long-protocol was not different from that of GnRH-antagonist protocol. Data on patients’ demographic and infertility treatment-related characteristics are represented in Table 1.</div>
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In fresh embryo cycles, there were less amount of gonadotropin and few days of OHSS for GnRH-antagonist protocols. Therefore, the total cost was lower in GnRH-antagonist protocols (average cost $16970.85) than in GnRH-agonist long-protocol (average cost $19902.24) in fresh embryo cycles. In fresh embryo transfer cycles, there was no statistical difference in the ongoing pregnancy rate between the two protocols, therefore, the minimum cost method was used for analysis. When the cumulative ongoing pregnancy rate was taken into account in the long protocol group, the number of frozen embryo transfer cycle was higher, leading to a higher cumulative ongoing pregnancy rate, and consequently a higher cost. Using cost-effectiveness analysis, the mean costs per ongoing pregnancy were estimated at $8176.76 and at $7595.28 with GnRH-antagonist protocols and GnRH-agonist long-protocols, respectively. The incremental cost-effectiveness ratio (ICER) is an evaluation index commonly used in economics research. It refers to the ratio of cost difference and effect difference between different protocols. In this study, the ICER for GnRH-agonist long-protocol versus GnRH-antagonist protocol was estimated at $4375.95 for one more ongoing pregnancy (Table 4).</div>
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<span style="background-color: initial; font-weight: bolder;">Table 4 Cost-Effectiveness analysis of GnRH-antagonist protocol and GnRH-agonist long-protocol.</span></div>
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<tr style="border-top: 0px;"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">GnRH-antagonist protocol</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">4958.79</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.6065</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">8176.76</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">—</td></tr>
<tr style="border-top: 1px solid rgb(213, 213, 213);"><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">GnRH-agonist long-protocol</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">5438.12</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">0.716</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">7595.28</td><td class="u-text-left " style="border-right: 1px solid rgb(213, 213, 213); line-height: 1.15; margin: 0px; padding: 6px; vertical-align: middle;">4375.95</td></tr>
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Sensitivity analysis is to change the value of some parameters and analyze the degree of influence on the results. The cost of ovarian stimulation protocols account for the majority of the total costs<span style="font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 10" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-65558-0#ref-CR10" id="ref-link-section-d141997e1090" style="background-color: transparent; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Bouwmans, C. A. et al. A detailed cost analysis of in vitro fertilization and intracytoplasmic sperm injection treatment. Fertility and sterility 89, 331–341,
https://doi.org/10.1016/j.fertnstert.2007.03.003
(2008).">10</a></span>. In this study, the cost of drugs accounted for the largest proportion of the total cost of ovulation stimulation protocols. The reliability of the results was assessed by sensitivity analysis with drug cost fluctuation. The results of sensitivity analysis were consistent with the results of cost-effectiveness analysis, the cost per ongoing pregnancy in the GnRH-agonist long-protocol was lower than that in the GnRH-antagonist protocol, which means the results are stable.</div>
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In this study, we used an economics research method to evaluate the cost-effectiveness of the GnRH-agonist long-protocol and the GnRH-antagonist protocol for IVF. We found that in the fresh embryo transplantation cycle, there was no significant difference in the ongoing pregnancy rate between the GnRH-agonist long-protocol and the GnRH-antagonist protocol, while the cost in the GnRH-antagonist protocol was lower, so GnRH-antagonist protocol has advantage according to the principle of cost-minimization analysis of pharmacoeconomics. When considering the cumulative ongoing pregnancy rate after each ovarian stimulation, the cumulative ongoing pregnancy rate and cost in the GnRH-agonist long-protocol were higher than the GnRH-antagonist protocol. Using the cost-effectiveness analysis method, it was found that the average cost of each ongoing pregnancy in the GnRH-agonist long-protocol was lower than the GnRH-antagonist protocol. Therefore, the GnRH-agonist long-protocol is more cost-effective.</div>
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<span style="font-size: 18px;">Studies have shown that one of the primary reasons for dropout from infertility treatment is economic burdens</span><span style="font-size: 13.5px;"> </span><span style="font-size: 18px;">. China’s medical system does not provide insurance coverage for infertility diagnosis and treatment</span><span style="font-size: 18px;">. It can be an enormous economic burden to patients seeking ART. Therefore, no matter from the perspective of patients, or from the perspective of medical resource allocation, it is necessary to carry out economic analysis on IVF and consider the cost and effect of each step. Although GnRH-agonist long-protocols and GnRH-antagonist protocols have been widely used in IVF, there is still an ongoing debate about the results of the two protocols. Orvieto</span><span style="font-size: 18px;"> and Grow</span><span style="font-size: 18px;"> found the GnRH-agonist protocol has a superiority over the GnRH-antagonist protocol in live birth rate. Some studies also found no significant difference in the rates of live births or ongoing pregnancies between the two protocols</span><span style="font-size: 18px;">. However, these studies are only from the perspective of clinical results, not from the perspective of economics. In our research, we hope to be able to focus more on evaluating the cost-effectiveness of both protocols, not just the clinical outcomes, which are just a link in the evaluation. After consulting the literature, we found that there have been little economic studies on the two different ovarian stimulation protocols used in IVF. Wei Pan</span><span style="font-size: 18px;"> </span><i style="font-size: 18px;">et al</i><span style="font-size: 18px;">. conducted a retrospective analysis of the cost-effectiveness of GnRH-a protocols, GnRH-ant protocols and GnRH-a ultra-long protocols. They used the live birth rate as one outcome of the study. However, it is difficult to calculate the cost throughout pregnancy. In order to ensure that the results are more reliable, we used the ongoing pregnancy rate as the end point of this economic study.</span></div>
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The cost of IVF treatment for infertility (the cost per ongoing pregnancy) in this study was higher than the average hospitalization cost for 30 diseases in 2018 according to the national bureau of statistics. IVF is a complex process involving ovarian stimulation, ovum retrieval, fertilization, embryo transfer and other processes. In addition to the cost of these processes, the total cycle costs of IVF should also include the transportation costs, lost wages and the cost of treating OHSS. However, it is difficult to accurately assess the transportation costs and lost wages and these indirect cost consisted a small percentage of the total costs, many studies did not include them in the total cycle cost analysis. But, from the perspective of patients, these costs are indirect medically, yet direct economically to patients, so they are also included in this study. OHSS is a serious complication of IVF, and the treatment is expensive, which directly affects the total cycle costs. Studies have shown that the incidence of OHSS in the GnRH-antagonist protocol is lower than that in the GnRH-agonist long-protocol. Accordingly, the cost of treating OHSS in the GnRH-antagonist protocol is lower, resulting in reduction in the total cycle cost. This may be one of the reasons why the cost in GnRH-antagonist protocol is lower than the GnRH-agonist long-protocol in the fresh embryo transfer cycle.</div>
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Economic analysis of IVF is still challenging, and there is no unified view on the result indicators of the analysis. Existing economic studies often used ongoing pregnancy, live birth rates or quality-adjusted life years (QALYs) as result of the study. However, it is worth considering that QALYs of both husband and wife or child is used when taking QALYs as a result of IVF. Toftager <i>et al</i>. found that quality of life and psychosocial and physical well-being of patients used the GnRH-antagonist protocol was better than that used the GnRH-agonist protocol. But these are hard to quantify in terms of costs. And it’s difficult to calculate the impact on families and society of obtaining a healthy baby by IVF. Considering the different incidence of related complications during pregnancy, the treatment costs and nursing costs vary greatly. Therefore, in this study, we used the cumulative ongoing pregnancy rate as the effect of the study.</div>
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There are many economic analysis methods, but among the existing studies on economics in ART, 84% are cost-effectiveness analysis and 48% are model-based studies<span style="font-size: 13.5px; line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;"><a aria-label="Reference 11" data-test="citation-ref" data-track-action="reference anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-65558-0#ref-CR11" id="ref-link-section-d141997e1178" style="background-color: transparent; color: #006699; overflow-wrap: break-word; text-decoration-line: none; vertical-align: baseline; word-break: break-word;" title="Moolenaar, L. M. et al. Economic evaluation studies in reproductive medicine: a systematic review of methodologic quality. Fertility and sterility 99, 1689–1694,
https://doi.org/10.1016/j.fertnstert.2012.12.045
(2013).">11</a></span>. Economic model carried out before a trial is particularly useful in reducing unnecessary waste of research resources in evaluating techniques and interventions and improving the quality and efficiency of the research. In our study, we developed a decision-tree model to evaluate the cost-effectiveness of the GnRH-agonist long-protocol and GnRH-antagonist protocol. The relevant transfer probability in the model was calculated using the data of infertile patients in the reproductive center of our hospital. In addition, we have made some reasonable assumptions for analysis, as shown in Table 2.</div>
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This is a retrospective study on the data available from infertility diagnosis and treatment in our reproductive center. The relevant probability and costs in the model are calculated based on the data of our single center. These results may differ from those of other centers, but can provide some guidance for patients and clinicians. In the future, large samples, multi-center prospective randomized controlled trials are needed to more thoroughly explore more economical and effective treatment protocols.</div>
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In conclusion, if fresh embryo transfers are considered, the pregnancy outcomes between GnRH-agonist long protocols and GnRH-antagonist protocols are similar, but GnRH-antagonist protocols have lower cost. Therefore, in the fresh embryo transfer cycle, the GnRH-antagonist protocol has economic advantage and is worth recommending. However, GnRH-agonist long-protocol have higher success rates and higher costs when cumulative ongoing pregnancy rates are taken into account. The cost per ongoing pregnancy in the GnRH-agonist long-protocol cycles was lower than that in the GnRH-antagonist protocol cycles. Thus, cost-effectiveness analysis shows that the GnRH-agonist long-protocol is more cost-effective than the GnRH-antagonist protocol and may represent a cost-effective option from the perspective of patients. However, further large sample sizes and multi-center randomized controlled trials are needed.</div>
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Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-70791722363311976102020-06-07T07:43:00.001-07:002020-06-07T07:43:30.236-07:00COVID-19: HYDROXYCHLOROQUINE NEWS UPDATE<div class="separator" style="clear: both; text-align: center;">
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COVID-19: HYDROXYCHLOROQUINE NEWS UPDATE</h2>
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<b><a data-saferedirecturl="https://www.google.com/url?q=https://nature.us17.list-manage.com/track/click?u%3D2c6057c528fdc6f73fa196d9d%26id%3D863f2c9cab%26e%3D48f7cc2c04&source=gmail&ust=1591627125613000&usg=AFQjCNEfUxvHIgimYB1XZqUVb8Fu3q1YCA" href="https://nature.us17.list-manage.com/track/click?u=2c6057c528fdc6f73fa196d9d&id=863f2c9cab&e=48f7cc2c04" style="color: #1155cc; text-decoration-line: none;" target="_blank">India doubles down on controversial drug</a></b><br />India’s government has <a data-saferedirecturl="https://www.google.com/url?q=https://nature.us17.list-manage.com/track/click?u%3D2c6057c528fdc6f73fa196d9d%26id%3Df7e74f599e%26e%3D48f7cc2c04&source=gmail&ust=1591627125613000&usg=AFQjCNFqf1Wkly9I0FtxhjKHR73gdtGl2A" href="https://nature.us17.list-manage.com/track/click?u=2c6057c528fdc6f73fa196d9d&id=f7e74f599e&e=48f7cc2c04" style="color: #1155cc; text-decoration-line: none;" target="_blank">extended its recommendation that frontline workers take the malaria drug hydroxychloroquine</a> to prevent coronavirus infections. Scientists have criticised the government for issuing advice on the basis of unpublished data and a published study that was not designed to test whether the drug actually prevents infection. Doctors point to cases where the government’s advice has contributed to widespread use of hydroxychloroquine, despite possible side effects.<br /><br /><br /><b><a data-saferedirecturl="https://www.google.com/url?q=https://nature.us17.list-manage.com/track/click?u%3D2c6057c528fdc6f73fa196d9d%26id%3De1cc6c8cf8%26e%3D48f7cc2c04&source=gmail&ust=1591627125613000&usg=AFQjCNHGu7bae1gXx5BaDKfUdC-2VEHUFg" href="https://nature.us17.list-manage.com/track/click?u=2c6057c528fdc6f73fa196d9d&id=e1cc6c8cf8&e=48f7cc2c04" style="color: #1155cc; text-decoration-line: none;" target="_blank">WHO restarts hydroxychloroquine trial</a></b><br />The <a data-saferedirecturl="https://www.google.com/url?q=https://nature.us17.list-manage.com/track/click?u%3D2c6057c528fdc6f73fa196d9d%26id%3Ded300debec%26e%3D48f7cc2c04&source=gmail&ust=1591627125613000&usg=AFQjCNEhHwCOvKhvjwnzzr7QlSU0qlBnvg" href="https://nature.us17.list-manage.com/track/click?u=2c6057c528fdc6f73fa196d9d&id=ed300debec&e=48f7cc2c04" style="color: #1155cc; text-decoration-line: none;" target="_blank">World Health Organization (WHO) has resumed testing hydroxychloroquine as a COVID-19 treatment</a>. The drug was temporarily removed from the WHO’s global Solidarity trial because of safety concerns raised by a large observational study published in <i>The Lancet</i>. Yesterday, the journal issued an <a data-saferedirecturl="https://www.google.com/url?q=https://nature.us17.list-manage.com/track/click?u%3D2c6057c528fdc6f73fa196d9d%26id%3D26d7500a39%26e%3D48f7cc2c04&source=gmail&ust=1591627125613000&usg=AFQjCNEWMuJvTis6wwXTPh5I_o_d8R4AOQ" href="https://nature.us17.list-manage.com/track/click?u=2c6057c528fdc6f73fa196d9d&id=26d7500a39&e=48f7cc2c04" style="color: #1155cc; text-decoration-line: none;" target="_blank">expression of concern</a>, and noted that an independent audit of the data has been commissioned.<br /><br /><b><a data-saferedirecturl="https://www.google.com/url?q=https://nature.us17.list-manage.com/track/click?u%3D2c6057c528fdc6f73fa196d9d%26id%3D5182598498%26e%3D48f7cc2c04&source=gmail&ust=1591627125613000&usg=AFQjCNHcCZD6aX_aZOMhvgysKaGOWztgng" href="https://nature.us17.list-manage.com/track/click?u=2c6057c528fdc6f73fa196d9d&id=5182598498&e=48f7cc2c04" style="color: #1155cc; text-decoration-line: none;" target="_blank">No evidence for hydroxychloroquine protection</a></b><br />A large clinical trial has found <a data-saferedirecturl="https://www.google.com/url?q=https://nature.us17.list-manage.com/track/click?u%3D2c6057c528fdc6f73fa196d9d%26id%3D740c803faa%26e%3D48f7cc2c04&source=gmail&ust=1591627125613000&usg=AFQjCNG8yrhbx8gb3DTpf7SICZ0IbaHuZQ" href="https://nature.us17.list-manage.com/track/click?u=2c6057c528fdc6f73fa196d9d&id=740c803faa&e=48f7cc2c04" style="color: #1155cc; text-decoration-line: none;" target="_blank">no evidence that hydroxychloroquine protects people from COVID-19</a>. The gold-standard trial randomly assigned 821 people to take either hydroxychloroquine or a placebo within 4 days of exposure to SARS-CoV-2. There was no statistically significant difference between the two groups in the number of people who developed COVID-19 within two weeks, but those taking the drug did report more side effects than did those taking the placebo. People were not tested unless they showed symptoms, so the study doesn’t take asymptomatic cases into account.<br /><a data-saferedirecturl="https://www.google.com/url?q=https://nature.us17.list-manage.com/track/click?u%3D2c6057c528fdc6f73fa196d9d%26id%3D8bb84253bf%26e%3D48f7cc2c04&source=gmail&ust=1591627125613000&usg=AFQjCNEQYiub6fs_R_PZyQK0JKnXWDsxpw" href="https://nature.us17.list-manage.com/track/click?u=2c6057c528fdc6f73fa196d9d&id=8bb84253bf&e=48f7cc2c04" style="color: #1155cc; text-decoration-line: none;" target="_blank">NPR | </a><br />Reference: <a data-saferedirecturl="https://www.google.com/url?q=https://nature.us17.list-manage.com/track/click?u%3D2c6057c528fdc6f73fa196d9d%26id%3D31593dc27f%26e%3D48f7cc2c04&source=gmail&ust=1591627125613000&usg=AFQjCNFKlSu6UWz3oz_hfrLSSMe_0_-Rxw" href="https://nature.us17.list-manage.com/track/click?u=2c6057c528fdc6f73fa196d9d&id=31593dc27f&e=48f7cc2c04" style="color: #1155cc; text-decoration-line: none;" target="_blank"><i>New England Journal of Medicine</i></a><a data-saferedirecturl="https://www.google.com/url?q=https://nature.us17.list-manage.com/track/click?u%3D2c6057c528fdc6f73fa196d9d%26id%3Dcd3894e778%26e%3D48f7cc2c04&source=gmail&ust=1591627125613000&usg=AFQjCNE_xmIbaHmcFg0M5agZxY_ogtPEZg" href="https://nature.us17.list-manage.com/track/click?u=2c6057c528fdc6f73fa196d9d&id=cd3894e778&e=48f7cc2c04" style="color: #1155cc; text-decoration-line: none;" target="_blank"> paper</a></div>
Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com0tag:blogger.com,1999:blog-204734490966806252.post-51573696947110744832020-04-13T19:00:00.002-07:002020-04-13T19:00:53.675-07:00Helping Hands for Covid 19<div class="separator" style="clear: both; text-align: center;">
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Dear all,<br />
As the Covid 19 has been spreed in all over the World, poor peolples and farmers are in more trouble. The farmers has their crops but all markets are down so mainly they are facing financial problems. Also in some villages there is lack of medical stores and due to lockdown they can not go outside to purchase the masks and sanitizers we are thinking to provide those things to them.<br />
<br />
As we are a community of them The company Arvigen Healthcare private limited has thought to help them by funding them as well as providing masks and sanitizers as no one is taking care of these people.<br />
<br />
So Arvigen Healthacare is taking interest in contrubution if you are also interested kindly do help by donating some amount whatever you want.<br />
<br />
Account details<br />
Arvigen healthcare private limited<br />
Icici bank Hadapsar Pune Branch<br />
<h2>
Accont no :337905000654</h2>
<h2>
IFSC code : ICIC0003379</h2>
<br />
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UPI Id: arvigenhealthcare@ybl</h2>
<br />Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com2tag:blogger.com,1999:blog-204734490966806252.post-65534586320639413572020-03-06T07:51:00.001-08:002020-03-06T08:18:20.758-08:00Sperm enrichment from poor semen samples by double density gradient centrifugation in combination with swim-up for IVF cycles<div class="c-article-header" style="margin: 0px 0px 40px; padding: 0px;">
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Sperm enrichment from poor semen samples by double density gradient centrifugation in combination with swim-up for IVF cycles</span></h1>
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<li class="c-author-list__item" itemprop="author" itemscope="itemscope" itemtype="http://schema.org/Person" style="display: inline; margin-left: 0px; padding-right: 0px;"><span itemprop="name"><a class="js-no-scroll" data-corresp-id="c1" data-test="author-name" data-track-action="open author" data-track-category="article body" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#auth-8" style="vertical-align: baseline;">Li Chen<svg class="u-icon" height="16" width="16"><use xlink:href="#global-icon-email" xmlns:xlink="http://www.w3.org/1999/xlink"><svg id="global-icon-email" viewbox="0 0 18 18"><path d="M1.995 2h14.01A2 2 0 0118 4.006v9.988A2 2 0 0116.005 16H1.995A2 2 0 010 13.994V4.006A2 2 0 011.995 2zM1 13.994A1 1 0 001.995 15h14.01A1 1 0 0017 13.994V4.006A1 1 0 0016.005 3H1.995A1 1 0 001 4.006zM9 11L2 7V5.557l7 4 7-4V7z" fill-rule="evenodd"></path></svg></use></svg></a></span> </li>
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Abstract</span></h2>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Sperm preparation in IVF cycles using density gradient centrifugation (DGC) in combination with swim-up (SU) has been widely adopted in reproductive centres worldwide. It is a fact that the sperm recovery rate following one DGC from poor semen samples (showing liquefaction defects/containing too many unresolvable clots or rare sperm) is relatively low. Our results showed that double DGC (DDGC) is effective at increasing the sperm recovery rate from poor semen samples. However, DDGC may increase the mechanical stress of sperm, thereby potentially impairing embryo development. Therefore, it is necessary to evaluate the safety of using sperm prepared by DDGC/SU for IVF cycles. In this study, we retrospectively analysed the data generated from a total of 529 IVF cycles (from June 2017 to June 2018), and these IVF cycles contributed 622 transfer cycles (from June 2017 to December 2018) in Changzhou Maternal and Child Health Care Hospital. Of them, 306 IVF cycles and the related 355 transfer cycles (normal semen samples prepared by DGC/SU) were set as the normal group, while 223 IVF cycles and the related 267 transfer cycles (poor semen prepared by DDGC/SU) were set as the observation group. The main outcome measures, including the normal fertilization rate, top D3 embryo formation rate, blastocyte formation rate, clinical pregnancy rate and live birth rate, birth weight and duration of pregnancy, were compared between the two groups. Compared to semen in the DGC/SU group, semen in the DDGC/SU group showed increased levels of the DNA fragmentation index (DFI) and reduced sperm concentration, percentage of progressive motility (PR) sperm, and percentage of normal morphology sperm. The indicators reflecting <i>in vitro</i> embryo development and clinical outcomes were similar in the DGC/SU group and DDGC/SU group, including the normal fertilization rate, top D3 embryo formation rate, blastocyte formation rate, pregnancy rate, implantation rate, spontaneous abortion rate, live birth rate, birth weight and duration of pregnancy. Furthermore, we found that the 1PN zygote formation rate was significantly lower in the DDGC/SU group than that in the DGC/SU group. We concluded that oocytes fertilized by sperm from poor semen samples separated by DDGC/SU achieved the same outcomes as oocytes fertilized by sperm from normal semen separated by DGC/SU, suggesting that DDGC/SU is an effective and safe method of sperm enrichment for poor semen samples in IVF. The main contribution of the present study is the verification of the effectiveness of DDGC/SU in improving sperm recovery from poor semen samples and the safety of using sperm prepared by DDGC/SU for IVF.</span></div>
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Introduction</span></h2>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">In <i>in vitro</i> fertilization (IVF), two major semen preparation methods have been widely used for selecting sperm from semen, namely density gradient centrifugation (DGC) and swim-up (SU). Numerous studies have focused on investigating which method is better for sperm enrichment. However, studies from different groups have drawn inconsistent conclusions. Recent studies have suggested that DGC in combination with SU (DGC/SU) could be one of the most effective approaches. Most Chinese reproductive centres, including ours, have adopted DGC/SU for enriching sperm in IVF.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">In clinical sperm preparation, the sperm recovery rate following one DGC largely depends on semen quality. Generally speaking, the sperm recovery rate from poor semen samples (such as liquefaction defects, containing too many unresolvable clots or rare sperm) is far from satisfactory. In clinical settings, when purifying sperm from semen for conventional IVF, we often encounter the situation where only a small proportion of sperm from a poor semen sample was obtained after one DGC. We have to make a choice: ignore the remaining motile sperm in the semen or re-extract them. In our reproductive centre, if few sperm are obtained after one DGC, the sperm precipitation (visible or invisible) is collected, and the remaining semen undergoes a second round of DGC with the old gradient column. Actually, the sperm from the double consecutive DGC would be pooled together for the next swim-up procedure. Here, we defined the double consecutive DGC in combination with SU as DDGC/SU.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">It has been demonstrated that an increase in the centrifugation time or gravitational (g) force can increase the sperm recovery rate from equine semen but can also decrease sperm motility or quality due to the mechanical forces associated with centrifugation and excessive packing of the sperm. Studies using spermatozoa from the epididymis of rats and mice revealed that increasing the centrifugation force decreased sperm function. A previous study demonstrated that centrifugation stress reduces the responsiveness of the spermatozoa of pigs to a capacitation stimulus. Although the sperm of bull are relatively resistant to high centrifugal forces, it has been demonstrated that a reduction in centrifugation force has increased the fertilization rate <i>in vitro</i>. Studies investigating the influence of centrifugation on human sperm showed that centrifugation induced damage to human sperm. Indeed, in the abovementioned studies, either conventional centrifugation or DGC was used for sperm preparation. Although it is believed that DGC causes less damage to sperm than conventional centrifugation, centrifugation per se is harmful. Therefore, it raises the concern that mechanical stress induced by either the increased centrifugation force or additional centrifugation time will further increase sperm damage, which may potentially affect the process of fertilization or embryo development. However, studies correlating the mechanical stress of sperm with embryo development are lacking. Although it has been demonstrated that sperm processing by DGC and SU is effective at reducing sperm with DNA damage, which is thought to definitely adversely affect early embryo development, it remains elusive whether a second DGC will introduce new damage to sperm, thereby finally impairing embryo development.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Although the sperm from both the first and second DGC were gathered, the safety of DDGC/SU for sperm needs to be evaluated. In this study, we retrospectively analysed the data generated from patients from June 2017 to December 2018 in the reproductive centre of Changzhou Maternal and Child Health Care Hospital for IVF treatment. According to the method of sperm processing, patients were divided into two groups: the DGC/SU group and the DDGC/SU group. The data generated in this study were compared between the two groups.</span></div>
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Results</span></h2>
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A second DGC significantly increased the motile sperm recovery rate from poor semen samples</span></h3>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">To determine whether a second DGC will increase the recovered sperm from semen, we separated and collected the sperms from normal and poor semen samples by first and second DGCs, and the parameters of sperm from the first and second DGCs were analysed. The results showed that an amount of sperm could still be extracted from normal or poor semen samples by the second DGC. However, the relative total sperm count and the ratio of PR-sperm acquired by the second DGC to the first DGC were much higher from poor semen samples than those from normal semen samples (Fig. <a data-track-action="figure anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Fig1" style="overflow-wrap: break-word; vertical-align: baseline;">1A–C</a>). These results indicated that the second DGC effectively separated motile sperm from poor semen samples.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">A second DGC significantly improves the recovery rate of motile sperm from poor semen samples. Normal or poor semen samples underwent two consecutive DGCs (462 g for 15 min), (<span style="font-weight: bolder;">A</span>) Representative images of sperm precipitation after the first DGC (left tube) and second DGC (right tube). (<span style="font-weight: bolder;">B</span>) Ratio of the total sperm count in the second DGC to the first DGC. (<span style="font-weight: bolder;">C</span>) Ratio of PR sperm in the second DGC to first DGC. Normal semen samples vs. poor semen samples; ***P < 0.001.</span></div>
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Basic characteristics of patients</span></h3>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">The present study included a total of 529 IVF cycles, 306 cycles for the DGC/SU group and 223 cycles for DDGC/SU. The patient characteristics and cycle parameters were compared between the two groups as shown in Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Tab1" style="overflow-wrap: break-word; vertical-align: baseline;">1</a>.The percentage of primary infertility, infertility duration, paternal or maternal age and BMI, ovulation induction protocols, basal hormonal levels, total gonadotropin (Gn) use, duration of Gn use, endometrial thickness, average oocytes retrieved and MII oocytes were comparable in the DGC/SU and DDGC/SU groups (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Tab1" style="overflow-wrap: break-word; vertical-align: baseline;">1</a>). The DNA fragmentation index (DFI) was significant higher in the DDGC/SU group than that in the DGC/SU group, while the sperm concentration, the proportion of progressive motility (PR) and normal morphology sperm were lower in the DDGC/SU group (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Tab1" style="overflow-wrap: break-word; vertical-align: baseline;">1</a>). In addition, the proportion of male-infertility factors was significantly higher in the DDGC/SU group (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Tab1" style="overflow-wrap: break-word; vertical-align: baseline;">1</a>).</span></div>
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<figure style="margin: 0px;"><figcaption class="c-article-table__figcaption" style="line-height: 1.4; margin-bottom: 16px;"><span data-test="table-caption" id="Tab1" style="display: block; font-weight: bolder; min-height: 90px; padding-left: 160px; text-align: justify;"><span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Table 1 Patient characteristics and cycle parameters.</span></span></figcaption><div class="u-text-right u-hide-print" style="margin: 0px; padding: 0px; text-align: right;">
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<a class="c-article__pill-button" data-test="table-link" data-track-action="view table" data-track-category="article body" data-track-label="button" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y/tables/1" rel="nofollow" style="border-radius: 20px; border: 4px solid rgb(188, 210, 220); display: inline-block; font-weight: 700; line-height: 1.4; margin-bottom: 10px; overflow-wrap: break-word; padding: 8px 20px; transition: all 0.2s ease-out 0s, all 0.2s ease-out 0s; vertical-align: baseline;"><span style="background-color: white; color: black; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Full size table<svg class="u-icon" height="16" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"><svg id="global-icon-chevron-right" viewbox="0 0 16 16"><path d="M7.782 7L5.3 4.518c-.393-.392-.4-1.022-.02-1.403a1.001 1.001 0 011.417 0l4.176 4.177a1.001 1.001 0 010 1.416l-4.176 4.177a.991.991 0 01-1.4.016 1 1 0 01.003-1.42L7.782 9l1.013-.998z" fill-rule="evenodd"></path></svg></use></svg></span></a></div>
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Comparison of <i>in vitro</i> embryo developmental parameters between the DGC/SU and DDGC/SU groups</span></h3>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">The results showed that the rate of normal fertilization and the formation rate of top D3 embryos (grade I and II) from either 2PN or 1PN zygotes and blastocysts from either top or not top D3 embryos (grade III) were comparable in the DGC/SU group and DDGC/SU group (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Tab2" style="overflow-wrap: break-word; vertical-align: baseline;">2</a>). However, the 1PN zygote formation rate was significantly lower in DDGC/SU group than that in the DGC/SU group (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Tab2" style="overflow-wrap: break-word; vertical-align: baseline;">2</a>).</span></div>
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<figure style="margin: 0px;"><figcaption class="c-article-table__figcaption" style="line-height: 1.4; margin-bottom: 16px;"><span data-test="table-caption" id="Tab2" style="display: block; font-weight: bolder; min-height: 90px; padding-left: 160px; text-align: justify;"><span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Table 2 <i>In vitro</i> embryo developmental parameters.</span></span></figcaption><div class="u-text-right u-hide-print" style="margin: 0px; padding: 0px; text-align: right;">
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<a class="c-article__pill-button" data-test="table-link" data-track-action="view table" data-track-category="article body" data-track-label="button" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y/tables/2" rel="nofollow" style="border-radius: 20px; border: 4px solid rgb(188, 210, 220); display: inline-block; font-weight: 700; line-height: 1.4; margin-bottom: 10px; overflow-wrap: break-word; padding: 8px 20px; transition: all 0.2s ease-out 0s, all 0.2s ease-out 0s; vertical-align: baseline;"><span style="background-color: white; color: black; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Full size table<svg class="u-icon" height="16" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"><svg id="global-icon-chevron-right" viewbox="0 0 16 16"><path d="M7.782 7L5.3 4.518c-.393-.392-.4-1.022-.02-1.403a1.001 1.001 0 011.417 0l4.176 4.177a1.001 1.001 0 010 1.416l-4.176 4.177a.991.991 0 01-1.4.016 1 1 0 01.003-1.42L7.782 9l1.013-.998z" fill-rule="evenodd"></path></svg></use></svg></span></a></div>
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<h3 class="c-article__sub-heading u-h3" id="Sec6" style="margin: 0px 0px 8px; padding: 0px; text-align: justify;">
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Comparison of clinical outcomes of embryo transfer between DGC/SU and DDGC/SU groups</span></h3>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Given that the clinical outcomes for cleavage stage embryo transfer differ greatly from blastocyst transfer, we subdivided groups of DGC/SU and DDGC/SU into D3 embryo and blastocyte subgroups. The number of D3 embryos or blastocytes per transfer was similar in the DGC/SU and DDGC/SU groups (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Tab3" style="overflow-wrap: break-word; vertical-align: baseline;">3</a>). The pregnancy rate, spontaneous abortion rate, implantation rate, live birth rate, birth weight and duration of pregnancy from the transfer of either D3 embryos or blastocytes were comparable in the DGC/SU group and DDGC/SU group (Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Tab3" style="overflow-wrap: break-word; vertical-align: baseline;">3</a>).</span></div>
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<figure style="margin: 0px;"><figcaption class="c-article-table__figcaption" style="line-height: 1.4; margin-bottom: 16px;"><span data-test="table-caption" id="Tab3" style="display: block; font-weight: bolder; min-height: 90px; padding-left: 160px; text-align: justify;"><span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Table 3 Embryos transfer and clinical outcomes.</span></span></figcaption><div class="u-text-right u-hide-print" style="margin: 0px; padding: 0px; text-align: right;">
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<a class="c-article__pill-button" data-test="table-link" data-track-action="view table" data-track-category="article body" data-track-label="button" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y/tables/3" rel="nofollow" style="border-radius: 20px; border: 4px solid rgb(188, 210, 220); display: inline-block; font-weight: 700; line-height: 1.4; margin-bottom: 10px; overflow-wrap: break-word; padding: 8px 20px; transition: all 0.2s ease-out 0s, all 0.2s ease-out 0s; vertical-align: baseline;"><span style="background-color: white; color: black; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Full size table<svg class="u-icon" height="16" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"><svg id="global-icon-chevron-right" viewbox="0 0 16 16"><path d="M7.782 7L5.3 4.518c-.393-.392-.4-1.022-.02-1.403a1.001 1.001 0 011.417 0l4.176 4.177a1.001 1.001 0 010 1.416l-4.176 4.177a.991.991 0 01-1.4.016 1 1 0 01.003-1.42L7.782 9l1.013-.998z" fill-rule="evenodd"></path></svg></use></svg></span></a></div>
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</section><section aria-labelledby="Sec7"><div class="c-article-section" id="Sec7-section" style="clear: both; margin: 0px; padding: 0px;">
<h2 class="c-article-section__title u-h2 js-section-title js-c-reading-companion-sections-item" id="Sec7" style="border-bottom: 1px solid rgb(213, 213, 213); line-height: 1.4; margin: 0px 0px 8px; padding: 0px; text-align: justify;">
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Discussion</span></h2>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Our data showed that sperm preparation for IVF cycles by the strategy of DDGC/SU could significantly improve the sperm recovery rate from poor semen samples without adversely affecting the biological processes, including fertilization and <i>in vitro</i> and <i>in vivo</i> embryo development, suggesting that DDGC/SU could be a simple, effective and safe way to separate sperm from poor semen samples for IVF.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">A previous study demonstrated that additional centrifugation time or increased centrifugation force may increase the sperm recovery rate. Consistent with this finding, we found that additional centrifugation time increased the recovered sperm from both normal and poor semen samples. More importantly, additional centrifugation can separate more highly motile sperms from poor semen samples than from normal samples. Poor semen samples characterized by high viscosity, unresolved fibres or clots may result in more drag force to retard the downward movement of motile sperms during DGC. Additional centrifugation provided enough power that allowed the motile sperm thoroughly move through the entrapment. In normal semen, motile sperm can be easily separated by first DGC, and the second DGC resulted in the congregation of sperm with limited motile capacity, even immotile sperm. Therefore, a second DGC is more necessary and effective for the separation of sperm from poor semen samples.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">As mentioned above, the efficiency of separating sperm from poor semen samples which are characterized by liquefaction defects, containment of too many unresolvable clots or rare sperm after one DGC, is very low. In some cases, due to a low sperm recovery rate, the enriched sperm by one DGC from poor semen samples will not meet the requirement of insemination by conventional IVF, and ICSI would be the substitute. Although the safety of ICSI is generally accepted, a recent systematic review paper showed that compared with IVF-conceived children, ICSI-conceived children may be at increased risk of autism and intellectual impairment and suggested an uncertainty of the long term safety of ICSI. Furthermore, insemination by ICSI will increase the financial burden of patients. Therefore, poor semen samples processed by DDGC/SU help to reduce the total intracytoplasmic sperm injection (ICSI) cycles.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">In the present study, although the DFI of fresh semen samples was not taken into consideration when deciding whether the second centrifugation should be performed, we found that the DFI was significantly higher in the DGGC/SU group than that in the DGC/SU group. Previous studies demonstrated that the level of DFI negatively correlated with semen parameters, including the sperm concentration, sperm motility, and normal sperm morphology. Consistently, we found that the sperm concentration, percentage of PR sperm and normal sperm morphology were lower in the DDGC/SU group. In addition, the male-infertility factor ratio was significantly higher in the DDGC/SU group. The other characteristics of couples undergoing IVF cycles were comparable in the DGC/SU group and DDGC/SU group, including the age and BMI of males and females, type, duration and diagnosis of infertility, ovulation induction protocols, the total dose and duration of Gn use, endometrial thickness, average oocytes retrieved and MII oocytes. These results demonstrated that “poor” semen in the DDGC/SU group showed poor semen quality.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">It has been shown that sperm with a high level of DFI negatively correlated with the outcomes of IVF-ET. However, the findings from other studies suggested that DFI was not able to predict the outcomes of IVF. Although no consistent conclusion was made, it is at least clear that sperm with high-level DFI will not contribute to improving the outcomes of IVF cycles. In the present study, although a higher DFI of semen was present in the DDGC/SU group, the indicators including the normal oocyte fertilization rate, top D3 embryo formation rate, blastocyte formation rate, pregnancy rate, implantation rate, live birth rate, birthweight and duration of pregnancy from either the transfer of D3 embryos or blastocytes were comparable in the DGC/SU group and DDGC/SU group. It has also been reported that sperm DFI positively correlated with <span style="font-weight: bolder;">s</span>pontaneous abortion. In the present study, we found that the spontaneous abortion rate following the transfer of either D3 cleavage embryos or blastocytes was comparable between the two groups. A recent study indicated that only DFI over a “threshold” may increase the risk of <span style="font-weight: bolder;">s</span>pontaneous abortion and suggested that each laboratory may have its own threshold of DFI. Therefore, we speculated that although a significantly higher DFI was observed in the DDGC/SU group, the DFI may not reach the threshold to induce spontaneous abortion. These results demonstrated that sperm from poor quality semen samples with higher DFI by DDGC/SU for IVF achieved the same outcomes as sperm from normal semen by DGC/SU, suggesting that DDGC/SU is a safe method for sperm enrichment for IVF.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Previous studies in rodents showed that increased centrifugation force would damage the cell organelles and membranes of sperm. However, few studies have been conducted to further explore the effect of the sperm suffering excess centrifugation force on the outcomes of IVF in any species. Only one study showed that rat sperm with over centrifugation had no impact on the fertilization rate. In line with that, we also found that double centrifugation did not decrease the fertilization rate, nor the other indicators mentioned above. It is known that DNA, located in the head of sperm, is highly compacted, and the most important role that sperm plays in fertilization is to transfer its DNA into oocytes. In the present study, we speculated that increased centrifugation force was not likely to damage sperm DNA, but to cause limited membrane or organelle damage that was far from affecting sperm motile capacity.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">The formation of the 1PN zygote may be associated with several causes, including the fusion of male and female pronucleus, the asynchronous disappearance of male and female pronucleus, and only male or female pronucleus formation. Theoretically, the first 2 causes will not impair further embryo development while the latter 2 will. To our surprise, we found that poor semen processed by DDGC/SU significantly decreased the ratio of 1PN zygote formation without affecting the ratio of top D3 embryo formation derived from 1PN zygotes, when compared with DGC/SU for normal semen, suggesting that poor semen processed by DDGC/SU synchronously decreased all causes-resulted the formation of 1PN.</span></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">In summary, our study verified that DDGC could effectively improve motile sperm recovery from poor semen, and showed that sperm for IVF from poor quality semen samples processed by DDGC/SU were able to achieve the same outcomes as sperm from normal semen by DGC/SU, suggesting that DDGC/SU is an effective and safe method of the treatment for poor semen samples in IVF, that is, poor semen samples can be processed by DDGC/SU for IVF in the clinic.</span></div>
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</section><section aria-labelledby="Sec8"><div class="c-article-section" id="Sec8-section" style="clear: both; margin: 0px; padding: 0px;">
<h2 class="c-article-section__title u-h2 js-section-title js-c-reading-companion-sections-item" id="Sec8" style="border-bottom: 1px solid rgb(213, 213, 213); line-height: 1.4; margin: 0px 0px 8px; padding: 0px; text-align: justify;">
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Materials and Methods</span></h2>
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Study subjects</span></h3>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">The data generated from a total of 529 IVF cycles (from June 2017 to June 2018) and these IVF cycle-contributed 622 transfer cycles (from June 2017 to December 2018) in Changzhou Maternal and Child Health Care Hospital, were collected and analszed in this retrospective study. Subject to semen preparation, data from IVF cycles or ET cycles were divided into two groups: the DGC/SU group and the DDGC/SU group. All of the included patients read and signed the informed consent form. This retrospective study was approved by the Ethics Committee of Changzhou Maternal and Child Health Care Hospital and Nanjing Medical University. All the treatments in the present study were performed strictly in accordance with the Declaration of Helsinki for Medical Research.</span></div>
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Semen preparation</span></h3>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Patients with 2–5 days of abstinence ejaculated semen into a sterilized cup by masturbation. After liquefaction, 10 µl of semen sample was drawn on the slide and covered with a cover glass to roughly estimate sperm concentration and sperm motile parameters, including the percentage of progressive motility (PR) sperm, non-PR (NP) and immobile (IM) sperm, under a microscope with a magnification of 400X. The sperm concentration (10<span style="line-height: 0; position: relative; top: -0.5em; vertical-align: baseline;">6</span>/ml) was roughly estimated as sperm counts in a 400X visual field. The remaining sample was placed on a density gradient column consisting of 1.5 ml of lower layer and 1.5 ml of upper layer (Irvine, USA) and centrifuged at 462 g for 15 min. For DGC/SU, the sperm pellet was re-suspended in 3 ml of IVF plus medium (Vitrolife, Sweden) and washed by centrifugation at 370 g for 9 min. After that, the sperm pellet was re-suspended in 0.5 ml of IVF medium. Ten microliters of medium was used to roughly estimate PR sperm counts. Then, 0.5 ml of IVF medium was dropped gently and slowly on the surface of the sperm suspension along the inner tube wall, and the tube was placed at a vertical angle in an incubator at 37 °C and 5% CO<span style="bottom: -0.25em; line-height: 0; position: relative; vertical-align: baseline;">2</span> for approximately 1.5 h before insemination for the purpose of swim up. For DDGC/SU, after the transfer of the sperm pellet obtained from the first DGC, the density gradient column was centrifuged immediately for another round. The sperm pellet from the second DGC was then mixed with the sperm pellet from the first DGC. The remaining procedures were the same as the abovementioned DGC/SU. A work flow of DDGC/SU is described in Fig. <a data-track-action="figure anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Fig2" style="overflow-wrap: break-word; vertical-align: baseline;">2</a>. For insemination, a micropipettor was used to transfer the sperm into the culture drop of the oocyte.</span></div>
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<figure style="margin: 0px;"><figcaption><span class="c-article-section__figure-caption" data-test="figure-caption-text" id="Fig2" style="display: block; font-weight: bolder; margin-bottom: 8px; text-align: justify;"><span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Figure 2</span></span></figcaption><div class="c-article-section__figure-content" style="margin: 0px 0px 16px; padding: 0px;">
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<a class="c-article-section__figure-link" data-test="img-link" data-track-action="view figure" data-track-category="article body" data-track-label="image" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y/figures/2" rel="nofollow" style="display: inline-block; margin: 0px 0px 16px; max-width: 100%; overflow-wrap: break-word; padding: 0px; vertical-align: baseline;"><picture><span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;"><source srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-59347-y/MediaObjects/41598_2020_59347_Fig2_HTML.png?as=webp" type="image/webp"></source><img alt="figure2" aria-describedby="figure-2-desc" loading="lazy" src="https://media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41598-020-59347-y/MediaObjects/41598_2020_59347_Fig2_HTML.png" style="border: 0px; height: auto; max-width: 100%; vertical-align: middle;" /></span></picture></a></div>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Work flow of DDGC. A total of 6 consecutive steps were involved in the entire process. Briefly, semen was placed on the density gradient column in the tube for the first DGC. After the first DGC, sperm precipitation was transferred into IVF medium. The semen in the tube on the column was subject to the second DGC. The sperm precipitation from the second DGC was also transferred to the abovementioned IVF medium. The sperm were then washed by centrifugation and re-suspended in 0.5 ml of IVF medium. Finally, sperm underwent swim-up procedure.</span></div>
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<a class="c-article__pill-button" data-test="article-link" data-track-action="view figure" data-track-category="article body" data-track-dest="link:Figure2 Full size image" data-track-label="button" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y/figures/2" rel="nofollow" style="border-radius: 20px; border: 4px solid rgb(188, 210, 220); display: inline-block; font-weight: 700; line-height: 1.4; margin-bottom: 10px; overflow-wrap: break-word; padding: 8px 20px; transition: all 0.2s ease-out 0s, all 0.2s ease-out 0s; vertical-align: baseline;"><span style="background-color: white; color: black; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Full size image<svg class="u-icon" height="16" width="16"><use xlink:href="#global-icon-chevron-right" xmlns:xlink="http://www.w3.org/1999/xlink"><svg id="global-icon-chevron-right" viewbox="0 0 16 16"><path d="M7.782 7L5.3 4.518c-.393-.392-.4-1.022-.02-1.403a1.001 1.001 0 011.417 0l4.176 4.177a1.001 1.001 0 010 1.416l-4.176 4.177a.991.991 0 01-1.4.016 1 1 0 01.003-1.42L7.782 9l1.013-.998z" fill-rule="evenodd"></path></svg></use></svg></span></a></div>
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Inclusive criteria of semen for DDGC/SU</span></h3>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">In this study, semen were processed by DDGC/SU if (a) semen showed liquefaction defects, or contained too many unresolvable clots/fibres, or contained rare but high motile sperm, and (b) the sperm pellet after the first DGC was invisible or almost invisible.</span></div>
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Computer-aided semen analysis system (CASA) analysis</span></h3>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">Ten microliters of semen was added to the chamber of slides, and after a warm-up (10 min), the slide was analysed by CASA. The sperm concentration and motility-related parameters were recorded. The results of semen quality parameters presented in Table <a data-track-action="table anchor" data-track-label="link" data-track="click" href="https://www.nature.com/articles/s41598-020-59347-y#Tab1" style="overflow-wrap: break-word; vertical-align: baseline;">1</a> and results 1, including the sperm concentration, total sperm count and ratio of PR sperm, were determined by CASA analysis.</span></div>
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IVF procedures</span></h3>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">More than half of the patients received ovarian stimulation with GnRH analogues, and the rest were treated without GnRH analogues for controlled ovarian hyperstimulation. Insemination was performed in IVF plus medium (Vitrolife, Sweden) using conventional IVF. Approximately 16–18 h after insemination, oocytes with 1 or 2 visible pronuclei were further cultured in G1 plus medium (Vitrolife, Sweden). On day 3 of culture, embryos were scored as grade I, II, III or VI based on the number, size, and shape of blastomeres and their degree of fragmentation, as previously described. Grade VI embryos were discarded. D3 embryos were subjected to transfer, frozen by vitrification technology or expanded in G2-plus culture medium (Vitrolife, Sweden). Blastocysts from day 5 or 6 were scored using the system of Gardner. The usable blastocysts were frozen by vitrification technology or transferred.</span></div>
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Statistical analysis</span></h3>
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<span style="background-color: white; font-family: "arial" , "helvetica" , sans-serif; font-size: large;">All analyses were performed by using GraphPad prism software (version 5). The chi-square test was used to compare the data of the constituent ratio. If continuous variables were normally distributed, Student’s t-test was used. If not, the Mann–Whitney U test was used. P < 0.05 was considered statistically significant.</span></div>
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Nanasaheb Nagarehttp://www.blogger.com/profile/04314943943590071313noreply@blogger.com1