bluuebunny/crossref_metadata_2025 huggingface dataset:
P-034 Rate of testicular histology failure in predicting successful testicular sperm extraction in Non-Obstructive Azoospermia
Authors: Tondo F , Albani E , Arruzzolo L , Castellano S , Cesana A , Smeraldi A , Levi Setti P.E
Published: June 2022

Study question

Does testicular histology predict successful or unsuccessful TEsticular Sperm Extraction (TESE) in Non-Obstructive Azoospermia (NOA) patients?

Summary answer

Testicular histology failed to predict successful TESE in 1 of 3 NOA patients.

What is known already

The management of patients with Non-Obstructive Azoospermia (NOA) involves TEsticular Sperm Extraction (TESE) combined with IntraCytoplasmic Sperm Injection (ICSI). Sperm retrieval is successful in up to 50% of men with NOA; however, there is no single clinical finding or investigation that can accurately predict a positive outcome. Previous studies have concluded that testicular histology is the best predictor of a successful or unsuccessful TESE.

Study design, size, duration

This is a retrospective study of 525 patients who underwent TESE between January 2018 and December 2020 in Humanitas Fertility Center.

Participants/materials, setting, methods

The cohort was dived in five groups: 287 NOA, 95 necrozoospermia, 18 anejaculation, 1 testicular trauma and 124 Obstructive Azoospermia (OA) patients.

Main results and the role of chance

Sperm was retrieved and cryopreserved in 218/287 patients with NOA (75,95%), 55/95 in necrozoospermia (57,89%), 18/18 anejaculation (100%), 1/1 testicular trauma (100%) and 120/124 in OA patients (96,77%). Interesting that, when we compare sperm recovery data through TESE with the histological assessment (HA) of corresponding testicular biopsy, we found discordance against HA. In particular we retrieved spermatozoa in 218 NOA patients but in 74 of them HA failed to detect them (33,94%). Moreover HA failed to detect spermatozoa in 8/55 (14,54%) in case of necrozoospermia, 1/18 (5,55%) in anejaculation and 10/120 (8,33%) in OA patients.

Limitations, reasons for caution

Testis's heterogeneity is a limitation of the study.

Wider implications of the findings

We found a discordance when HA is compared with sperm recovery data of corresponding testicular biopsy. This data suggest that in case of testicular diagnostics histology in 33,94% of NOA cases there is a wrong diagnosis and 1 of 3 patients it doesn’t retrieve and cryopreserve spermatozoa when it could.

Trial registration number

N A
P-162 GDF9 concentration in embryo culture medium is linked to human embryo quality and viability
Authors: Li C , Jingyu L , Guoning H
Published: June 2022

Study question

Is the GDF9 concentration in Day 3 human embryo culture medium linked to embryo quality and clinical outcome?

Summary answer

The embryo quality and clinical outcome appears to be linked to the GDF9 concentration in Day 3 culture medium.

What is known already

Previous studies have demonstrated that the GDF9 level in follicular fluid and cumulus granulosa cells can be used as biomarker for predicting embryo quality.

Study design, size, duration

In study 1, a total of 280 embryos from 70 patients who obtained at least 4 embryos with 6-10 blastomeres (2 transferable and 2 non-transferable embryos) at Day 3 were enrolled. The corresponding GDF9 concentration in culture media were compared between transferable and un-transferable embryos and also compared among good, fair and poor quality embryos. In study 2, a total of 119 embryos from 61 patients (29 implanted and 32 non-implanted patients) were enrolled.

Participants/materials, setting, methods

The expression pattern of GDF9 in human embryos was investigated using Q-PCR and immunofuorescence. GDF9 concentration in embryo culture media was quantified by ELISA assay. Statistical analyses defined correlations between GDF9 concentration and embryo quality, in study 1, and implantation and live birth, in study 2.

Main results and the role of chance

GDF9 mRNA and protein were detected from human oocytes to eight-cell embryos and displayed a slow decreasing trend. In study 1, GDF9 concentration in culture medium is lower for transferable embryos compared with non-transferable embryos (331 pg/mL (quartiles: 442, 664 pg/mL) vs. 518 pg/mL (quartiles: 328, 1086 pg/mL), P < 0.001), and increased commensurate with the diminution of the embryo quality (P < 0.001). In study 2, signifcantly lower GDF9 concentration was detected for implanted embryos than non-implanted embryos (331 pg/mL (quartiles: 156, 665 pg/mL) vs. 518 pg/mL (quartiles: 328, 1086 pg/mL), P < 0.001). The same trend was found between the embryos that led to live birth and those that failed.

Limitations, reasons for caution

Further randomized controlled trials are needed for exploring whether GDF9 assay of spent embryo culture medium can improve the clinical outcome in IVF.

Wider implications of the findings

GDF9 in culture medium has the potential to be a non-invasive biomarker for embryo selection.

Trial registration number

not applicable
P-204 Transcriptome analysis of cell-free RNAs in embryonic culture medium reveals their potentials on non-invasive testing
Authors: Zhou Q , Zhongzhen L , Tingyu Y , Wen-jing W
Published: June 2022

Study question

Could cell-free RNA (cfRNA) in culture medium be non-invasive strategy to select in vitro fertilized (IVF) embryos?

Summary answer

CfRNAs in culture medium could be potential markers correlated with embryo qualities.

What is known already

Infertility is a critical component of reproductive health and couples with infertility may choose assisted reproductive technology (ART) to increase their chances of conception. However, the effectiveness remains low. One way to improve the outcome is to screen the embryo prior to implantation. Noninvasive testing opens new perspectives in analyzing embryo qualities and current pending includes microscopy images coupled with artificial intelligence and omics analysis of the spent blastocyst medium (SBM). Cell-free DNA in SBM could be used to identify the embryo ploidy and metabolomic/proteomic markers could be correlated with embryo potential, while the research of cfRNA is lacking.

Study design, size, duration

We developed a new library method of polyadenylation ligation-mediated sequencing (PALM-Seq) to capture the low amount of cfRNA and collected twelve culture mediums of in vitro embryos on Day 3 and five samples of embryos on Day 5.

Participants/materials, setting, methods

We employed PALM-Seq on culture medium of embryos generated by IVF or Intracytoplasmic sperm injection (ICSI). Owing to the new method, we could comprehensively profile the cfRNAs, including miRNA, tRNA, piRNA, lncRNA and mRNA. The morphology and fragmentation of the recruited embryos were also recorded.

Main results and the role of chance

Generally, thousands of genes could be detected in all culture mediums and the number greatly increased in SBM. We identified several reported miRNA markers in our study, such as miR-21-5p and miR-92a-3p, while highly expressed miRNAs varied across samples and stages. We indeed found piRNA in mediums and several specific piRNAs had large amount of expression. In contrast, detected number and expression of tRNA were stable across samples. Besides of reported mRNA markers, we could profile the expression of all detected mRNAs and no differences were found between embryos with and without fragmentation. We found that the majority of mRNA fragments were shorter than 30 nucleotides. In addition, some granular-specific expressed mRNAs could be detected in medium on Day 3 and they decreased of medium on Day 5.

Limitations, reasons for caution

We have only conducted preliminary exploration in the culture medium of pre-implantation embryos and studies of larger sample size are needed.

Wider implications of the findings

Our research proved the feasibility of performing high-throughput sequencing of cfRNAs in SBM. It opens new perspectives on defining non-invasive markers and selection of embryos with the most potential. Our results also contribute a new insight on improving the treatment of infertility.

Trial registration number

2018YFC1004900
P-207 Molecular mechanisms of melatonin action in human reproductive system – implications for assisted reproduction
Authors: Nikolov G , Konakchieva R , Nikolaev G , Marinova E
Published: June 2022

Study question

What is the potential role of melatonin and melatonin receptors activation on the reproductive function in humans?

Summary answer

The presence of membrane melatonin receptors in male gametes and female hGLCs suggests a role for melatonin on motility performance of mature spermatozoa and oogenesis.

What is known already

The interest in the influence of the external environment on the reproductive performance in humans facilitates reevaluation of its main mediators in the body. One of the most important among them is melatonin circadian secretion by the pineal gland, a major synchronizer of the diurnal rest-activity cycle. Activation of melatonin-linked GPCR receptors by melatonin is coupled to inhibition of cAMP accumulation in target cells, which makes research on membrane receptor-mediated signaling in human reproductive system very attractive.

Study design, size, duration

Pilot study on the presence of MT1 and MT2 receptors in human spermatozoa and human granulosa luteinized cells (hGLCs) and the presence and localization of α- and ß-tubulin, melatonin receptors (MT1, MT2) and P450 aromatase in hGLCs.

Participants/materials, setting, methods

Human donor samples (spermatozoa and granulosa lutein cells (hGLC) were collected after informed consent signed from patients recruited at IVF clinics in Sofia. Two-step quantitative real time RT-PCR (qRT-PCR) was used to compare individual variations between donors. Selected proteins in samples and derived cell cultures like α- and ß-tubulin, melatonin receptors (MT1, MT2) and P450 aromatase were studied by Western blotting and localization visualized by confocal immune fluorescence.

Main results and the role of chance

PCR amplification of cDNA in the samples resulted in products of the expected size of 516 bp corresponding to the MTNR1A gene. In purified spermatozoa, intense expression of a MT1 receptor was found in the post-acrosomal region and the neck. In hGLC cultures up-regulation of melatonin receptors after in-vitro melatonin treatment was observed both by protein expression and densitometry of immune fuorescence.

Our results demonstrate the presence in situ of membrane melatonin receptor in competent human male gametes and female hGLCs. The observed localization of the receptor protein suggests a role for melatonin on motility performance of mature spermatozoa, which may have several important implications to assisted reproductive techniques.

Limitations, reasons for caution

The discrete functional responses and the transduction pathways linked to specific receptor activation by melatonin in human reproductive system have further to be elucidated.

Wider implications of the findings

Future studies could reveal the potential therapeutic role of Melatonin in infertility patients.

Trial registration number

Not applicable
P-223 Women aged 40 or older and non-male factor have the same reproductive outcome using intracytoplasmic sperm injection (ICSI) or conventional in vitro fertilization (cIVF)
Authors: Filardi P , De Martino E , Chiarello Sosa J.M , Gomez Peña M , Papayannis M , Figueroa C , Magri L , Oubiña A , Horton M , De Zuñiga I , Kopcow L , Sobral F , Bisioli C
Published: June 2022

Study question

Do women aged 40 or older have the same reproductive outcome using ICSI or cIVF when male factor infertility is not present?

Summary answer

Women aged 40 or older with non-male factor partners achieved similar reproductive outcomes either with ICSI or cIVF.

What is known already

Although initially proposed to treat couples with severe male factor infertility, ICSI has become the preferred tool to treat patient populations others than initially intended. Many studies and international forums, however, have alerted that its overuse is not free due to its safety, cost and time-consuming concerns. Moreover, it was noted that it does not increase clinical outcomes even in cases of compromised oocyte factor such as poor responder patients or few oocytes retrieved. However, there is still no information regarding its use in the group of patients of advanced reproductive age.

Study design, size, duration

Retrospective cohort study including 412 women aged 40 or older with non-male factor infertility performing ICSI or cIVF between May 2015 and October 2017.

Participants/materials, setting, methods

Patients underwent ICSI (n = 109) or cIVF (n = 303) according to medical indication. The decision to use ICSI in non-male factor patients was merely operational, with the objective of evaluating oocyte maturity.Those cycles with own oocytes, non-male factor and fresh embryo transfers were included. Semen samples were considered normal following the World Health Organization fifth edition sperm parameters values, and prepared by density gradient selection.

Main results and the role of chance

The primary outcome was clinical pregnancy rate. Secondary outcomes included ongoing pregnancy, miscarriage and implantation rates. Statistical significance was denoted by P < 0.05. The mean age of the women were 41.4 in the ICSI group and 41.6 years in the cIVF group, while the mean age for male couple was 43.6 and 41.6 respectively.

No differences between ICSI and cIVF groups were observed regarding clinical pregnancy [16.5% (18/109) vs. 20.8% (63/303)], ongoing pregnancy [9.2% (10/109) vs. 7.9% (24/303)], miscarriage [41.2% (7/17) vs. 46,5% (20/43)], and implantation rates [11.3% (17/151) vs. 9.9% (50/506)].

Limitations, reasons for caution

These results need confirmation with a bigger population size. As with any retrospective study, the potential for residual confounding exists.

Wider implications of the findings

These data suggest that ICSI offers no clinical benefit for women aged 40 and older with non-male factor infertility, beyond oocyte maturation classification.

Trial registration number

not applicable
P-284 Correlation between a new day 4 embryo scoring system and blastocyst development and pregnancy rate
Authors: Martello C , Andrade G , Frantz G , Pagnoncelli N , Dutra C , Rockenbach M , Donatti L , Frantz N
Published: June 2022

Study question

Does a new day 4 embryo scoring system predict blastocyst development and aneuploidies?

Summary answer

The new day 4 embryo scoring system correlates with blastocyst development, blastocyst morphology, day of blastulation, and pregnancy rate.

What is known already

A small number of studies propose different day 4 embryo grading systems. The majority of them report no differences in implantation and clinical pregnancy outcome between day 4 and blastocyst transfer. Moreover, only limited data exist on the association between day 4 grading systems, embryo development, and PGT-A results. In addition, none of the previous studies analyzed the totality of the different day 4 embryo morphology and their subsequent development.

Study design, size, duration

This is a single-center retrospective analysis from June 2019 to August 2021 including 762 embryos from 129 cycles. Day 4 embryos were graded A to J considering A the best morphology and were classified into 1, 2, 3, or 4 categories (1 being the more advanced development). Categories were compared among them to assess blastocyst development and aneuploidy rate.

Participants/materials, setting, methods

Data were obtained from medical records with a previously given consent. Patient characteristics such as age, body mass index (BMI), and anti-Mullerian hormone (AMH) were compared among the four categories using ANOVA with Multiple Comparisons. Blastocyst development, blastocyst morphology, day of blastulation, and pregnancy rate were compared among the categories using Chi-square contingency. All analyses considered a significant p-value of 0.05.

Main results and the role of chance

Younger women presented significantly more category 1 of day 4 embryos when compared with the other categories (p < 0.0001). BMI and AMH were similar among the categories (p = 0.3063 and p = 0.2533, respectively). There was a high correlation between day 4 categories and PGT-A blastocyst development (p < 0.0001), blastocyst morphology (p < 0.0001), and day of blastulation (p < 0.0001). Also, there was a significant correlation between pregnancy rates and the four categories (p = 0.0192). No correlation was observed when aneuploidy rate and level of aneuploidy were analyzed (p = 0.3614 and p = 0.1490, respectively).

Limitations, reasons for caution

The limited number of embryos analyzed is a limitation of the study, and additional data are being gathered. Multiple embryos of the same patients can be a cofounder of the study and more robust statistical analysis may overcome it.

Wider implications of the findings

The new day 4 embryo score may be useful for embryo selection to PGT-A biopsy. Moreover, the proposed score may help to add valuable information to select the best embryos for transfer.

Trial registration number

not applicable
P-353 Placenta previa in In Vitro Fertilization and unassisted pregnancies – is there a difference in perinatal outcomes and placental histology?
Authors: Barber E , Ganer Herman H , Kovo M , Tairy D , Schreiber L , Horowitz E , Weissman A , Weiner E , Raziel A
Published: June 2022

Study question

Why is In-vitro-fertilization (IVF) an independent risk factor for placenta previa (PP).

Summary answer

While PP in non-assisted pregnancies is probably associated with previous cesarean deliveries (CD), in IVF it is more “sporadic”, and may complicate any index pregnancy.

What is known already

PP is more prevalent following IVF pregnancies as compared with unassisted pregnancies, with an increased risk of three to six-fold according to some authors. The etiology for this increased risk in IVF pregnancies in unclear, and may be related to reproductive procedures performed or to subfertility characteristics which have led to IVF.

Study design, size, duration

A retrospective-cohort study of deliveries with PP between 2008 and 2021. A total of 182 pregnancies were included.

Participants/materials, setting, methods

Placental histopathology, obstetric and neonatal outcomes were compared between IVF and unassisted pregnancies. Included, were singleton deliveries complicated by PP at gestational weeks (GA)> 24. Placental pathology was obtained utilizing the well-accepted Amsterdam criteria.

Main results and the role of chance

Out of 182 pregnancies which were included - 23 IVF pregnancies (IVF group) and 159 in the unassisted pregnancies (Control group). The control group was characterized by higher gravidity (p = 0.007) and parity (p < 0.001), whereas the IVF group- by a higher rate of nulliparity (p < 0.001) and diabetes mellitus (p = 0.04). A higher rate of patients with past CDs was noted in unassisted pregnancies, although not statistically significance (38.9% versus 21.7%, p = 0.10). There was a trend for a lower placental weight in the control group, and a higher incidence of placental weight below the 10thpercentile in this group (47.8% versus 13.9%, p = 0.001). No differences were noted in maternal and fetal vascular lesions. Nevertheless, IVF and unassisted pregnancies entail similar perinatal outcomes in cases of PP.

Limitations, reasons for caution

A major limitation was our small sample size in the IVF group. Despite a power calculation, larger study groups would have possibly allowed for the demonstration of additional differences in outcomes, including previous cesarean deliveries and placenta accreta. Moreover, this limitation prevented us from matching against possible cofounders.

Wider implications of the findings

While a growing proportion of pregnancies worldwide are conceived by IVF, possible iatrogenic side effects should be studied. As PP is of clinical importance, it is essential to diagnose it on time, as well as study the mechanisms and risk factors behind it, which could possibly help with its prevention.

Trial registration number

0282-20-WOMC
P-368 Serum progesterone on the day of embryo transfer in stimulated cycles does not correlate with pregnancy outcomes
Authors: Rozen G , Rogers P , Polyakov A
Published: June 2022

Study question

Is the chance of positive hCG, clinical pregnancy or live birth in a fresh embryo transfer (ET) IVF cycle, affected by day of transfer serum progesterone (P4) level?

Summary answer

There is no correlation between P4 level on the day of ET and pregnancy outcomes.

What is known already

Data from studies of frozen–thawed embryo transfer cycles suggest that low as well as high P4 levels during the mid-luteal phase may decrease the chance of pregnancy. The altered P4 pattern may disrupt the endometrial maturation leading to asynchrony between embryonic development and endometrial receptivity, thereby, compromising implantation and early development of pregnancy.

Study design, size, duration

A monocentric retrospective study of prospectively collected data, of women undergoing IVF or ICSI treatment July 2019-July 2020, and having a fresh day 5 single embryo transfer (SET). 825 first and second stimulation cycles were included. Patients underwent a GnRH-antagonist protocol and were triggered for final oocyte maturation with hCG (OvidrelR). All patients received vaginal luteal support. The study population was unselected, treated with the unit’s usual stimulation protocols, representing an everyday patient cohort.

Participants/materials, setting, methods

Serum P4 levels on the day of ET were correlated with positive hCG, clinical pregnancy (CP) and live birth (LB) rates. Based on previously defined P4 thresholds, patients were divided into four P4 groups based on serum levels (P4<150 nmol/l, P4 150–250 nmol/l, P4 251–400 nmol/l and P4>400 nmol/l). The data was further interrogated using additional P4 cut offs, to confirm that there were no discernible differences based on P4 levels.

Main results and the role of chance

There was no level of P4 below or above which the chance of pregnancy was consistently reduced. The chance of a positive hCG-test in the whole cohort was 51%, (95% CI: 47.1-54) following blastocyst transfer and was not different across the four groups (50, 48.2, 51.1, 52%, P = 0.88). Likewise, there was no difference in CP 41.8% (CI 38.4-45.3%) and LB 30.4% (27.3-33.7%), between the four groups. No negative association between P4 and chance of pregnancy when the other thresholds, described above, were applied. All estimates were adjusted for maternal age, maternal BMI, eggs retrieved and number of eggs fertilised, with the outcome remaining unchanged.

Limitations, reasons for caution

The finding of no difference in pregnancy outcomes across P4 ranges in this study, only applies to fresh blastocyst transfer IVF cycles, using hCG trigger and vaginal LPS, and should be corroborated in future clinical trials. Also, there is possible inaccuracy in P4 levels due to diurnal variability.

Wider implications of the findings

Future studies are necessary to explore the diurnal variability, and its possible contribution to these results.

Trial registration number

retrospective study, institutional ethics approval
P-445 Recovery of metabolic activity and VEGF secretion after thawing of cryopreserved, slow frozen ovarian tissue
Authors: Einenkel R , Schallmoser A , Saenger N
Published: June 2022

Study question

Is slow-frozen ovarian cortical tissue limited in its metabolical activity after thawing, especially with respect to VEGF secretion?

Summary answer

Slow-frozen ovarian cortical tissue showed decreased metabolic activity and VEGF directly after thawing, but recovered in the course of 48h in in vitro culture.

What is known already

Cryopreservation of ovarian cortical tissue has become an important method in fertility preservation. Freezing and thawing processes had been optimized to preserve tissue viability. However, the major obstacle of retransplantation is the rapid angiogenesis to accomplish sufficient blood supply. As freezing and thawing entails constraints in cellular metabolic activity, we aimed to assess the metabolic activity and the secretion of the angiogenic factor VEGF of frozen-thawed ovarian cortical tissue over time.

Study design, size, duration

Ex vivo gained ovarian tissue was cultured over 48h. Thereby, non-frozen fresh tissue was compared to slow-frozen-thawed ovarian tissue.

Participants/materials, setting, methods

Provided a signed consent, biopsies of ovarian tissue from patients undergoing fertility preservation were cultured without freezing (n = 13) or after slow-freezing and thawing (n = 12). VEGF secretion during 48h was measured by ELISA. Applying CellTiterBlue assay, metabolic activity was assessed in biopsies without freezing or after thawing from the same patients (n = 4-7) after 0, 24 and 48h. Similarly, VEGF secretion was measured after 0, 24 and 48h as well (n = 7) to capture temporarily changes.

Main results and the role of chance

Although VEGF secretion varies among patients and biopsies, there was a significant lower VEGF content in supernatant from frozen-thawed tissue compared to fresh ovarian cortical tissue after 48h of culture (p < 0.01). Directly after thawing and 24h thereafter, metabolic activity was significantly reduced (p < 0.05) compared to fresh ovarian cortex. In the course of 48h, the metabolic activity recovered (p < 0.01). Similarly, VEGF secretion increased dramatically during 24h after thawing (p < 0.05).

Limitations, reasons for caution

This in vitro study might not resemble the metabolic and secretory recovery after retransplantation.

Wider implications of the findings

We showed that ovarian tissue recovers metabolically over 48h. This recovery included an increase in VEGF secretion. As latter induces angiogenesis a culture step after thawing might support VEGF secretion and angiogenesis. However, the recovery period could also be useful as a temporal bridge until sufficient supplies are available.

Trial registration number

not applicable
P-659 Does Preconception thyroid-stimulating hormone (TSH) levels affect the clinical outcomes of Infertile Women Undergoing Fresh Embryo Transfer Cycle?
Authors: Song G.Y , Park E.A , Lee J.H , Ryu H.E , Park S.J , Hur Y.J , Yu E.J , Lee K.A , Yoon T.K
Published: June 2022

Study question

Does Preconception TSH levels affect the clinical outcomes of Infertile Women Undergoing Fresh Embryo Transfer Cycle?

Summary answer

Among euthyroid patients, preconceptional TSH values in the high-normal range (between 2.5 and 4.0 mIU/L) are associated with adverse clinical outcomes.

What is known already

Serum thyroid-stimulating hormone (TSH) levels are routinely screened in women with infertility given the negative impact of thyroid disease on ovarian function and pregnancy is associated with poor outcomes.In fact, an underlying thyroid abnormality was found in 46% for women experiencing IVF failure and TSH levels were inversely proportional to the fertilization rate at IVF. While it is clear that pregnancies complicated by subclinical hypothyroidism in the fertile population have poor outcomes, the relevance of preconception subclinical hypothyroidism in infertile patients undergoing ART has not been thoroughly examined.

Study design, size, duration

Outcomes of all fresh IVF patients with an embryo transfer from from January 2019 to August 2021 within 1 yr of TSH screening at a our center were studied. IVF cycles using donor oocyte, TESE sperm, PGT, or those lacking follow-up were excluded. For patients with more than one Preconception TSH measurement prior IVF, the most recent measurement before the IVF cycle was used.

Participants/materials, setting, methods

Cycles were categorized into two groups based on preconceptional TSH values: Euthyroid (0.4–2.5 μIU/mL), subclinical hypothyroidism (>2.5-4.0 μIU/mL). All women underwent standard IVF protocols following usual individualized practice in our IVF clinic. Chi-square test was used to evaluate the differences between the patient cohorts.

Main results and the role of chance

A total of 2,669 fresh-ET cycles were reviewed retrospectively by dividing them into a euthyroid group (0.4-2.5 uIU/mL, n = 2397) and subclinical hypothyroidism group (2.5-4.0 uIU/mL, n=272). The overall biochemical pregnancy rate and clinical pregnancy rate was significantly lower in women with subclinical hypothyroidism (50.0% vs 43.4%, 40.0% vs 30.9%, p < 0.05). However, biochemical preganacy loss rate and the miscarriage rate was higher in women with subclinical hypothyroidism. (19.6% vs. 28.8%, 17.3% vs. 25.0%, p = 0.079)

Limitations, reasons for caution

This is a retrospective study and results must be confirmed on a well-designed randomized controlled study. Correlations must be considered with caution since the role of TSH is under study, especially when considering LBR outcome.

Wider implications of the findings

Among euthyroid patients, preconceptional TSH values in the high-normal range (between 2.5 and 4.0 mIU/L) are associated with adverse clinical outcomes. Our findings suggest that subclinical hypothyroidism may affect clinical outcomes.

Trial registration number

not applicable
P-723 Lack of seasonal influence on fresh IVF/ICSI treatment outcomes. A population based nationwide registry study
Authors: Carlsson Humla E , Bergh C , Akouri R , Tsiartas P
Published: June 2022

Study question

Is there a seasonal influence on fresh in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment outcomes?

Summary answer

Season does not seem to affect live birth rate, clinical pregnancy rate or miscarriage rate after fresh IVF/ICSI treatment.

What is known already

Seasonal variations in human natural conception and birth rates are well described. However, the impact of seasonal variation on IVF outcomes has not yet been clarified and conflicting reports have been published.

Study design, size, duration

Nationwide, register-based cohort study including all first time fresh IVF/ICSI treatments (n = 52 788) leading to oocyte pick up (OPU) in Sweden between 2009 and 2018.

Participants/materials, setting, methods

First time fresh IVF/ICSI cycles leading to OPU were identified in the National Quality Registry of Assisted Reproduction (Q-IVF). Data collected included patient characteristics, information about the treatment cycle and pregnancy outcome. The patients were divided into season subgroups based on the date for OPU. The primary outcomes of the study were live birth per OPU and per embryo transfer (ET), clinical pregnancy per ET and miscarriage per clinical pregnancy. Logistic regression was performed.

Main results and the role of chance

The impact of seasons on the outcome of first time fresh IVF/ICSI cycles leading to OPU was assessed. A statistically significant overall seasonal influence was seen on live birth rate per OPU in the unadjusted analysis (p =0.036), where cycles performed in summer showed lower live birth rate (OR 0.92, 95% CI 0.87-0.97, p =0.004) compared with the other seasons. After adjustment for relevant confounders the overall impact of seasons on live birth rate was no longer statistically significant (p =0.10). Although post-hoc analyses still showed lower live birth rate in summer compared with the other seasons (OR 0.93, 95% CI 0.88-0.99, p =0.019). The impact of seasons on clinical pregnancy rate, miscarriage rate and live birth rate was further assessed through the analysis of the first cycles where ET was performed. No overall statistically significant seasonal influence was seen on clinical pregnancy, miscarriage and live birth rates after adjustment for confounders.

Limitations, reasons for caution

Possible limitations are the retrospective design of the study and the lack of adjustment for other important confounders i.e. cause of infertility, not included in Q-IVF.

Wider implications of the findings

This large cohort study shows a lack of seasonal influence on live birth-, clinical pregnancy- and miscarriage rate after fresh IVF/ICSI treatment. In the clinical setting, season should not be taken into consideration when planning and performing fresh IVF/ICSI.

Trial registration number

non applicable
P-748 Cesarean scar defect does not predict embryo transfer outcomes: A retrospective cohort study
Authors: Gale J , Lanes A , Corran B , Bacal V , Nguyen V , Shmorgun D , Singh S.S , Walker M
Published: June 2022

Study question

Is the presence of a cesarean scar defect associated with live birth outcomes at the time of embryo transfer?

Summary answer

The presence of a cesarean scar defect is not predictive of live birth rates at the time of embryo transfer.

What is known already

Cesarean delivery is associated with subsequent impaired fertility, with lower pregnancy and live birth rates and longer inter-pregnancy intervals, compared to patients who delivered vaginally. Studies exploring pregnancy rates after IVF and embryo transfer also suggest lower pregnancy rates among patients with prior cesarean delivery. A postulated mechanism by which a prior cesarean delivery may impact fertility is the cesarean scar defect, and prior studies have shown a possible association between a scar defect and suboptimal outcomes.

Study design, size, duration

We retrospectively identified a patient cohort based out of the Ottawa Fertility Centre in Ottawa, Ontario, Canada, between 2013 and 2019.

Participants/materials, setting, methods

This retrospective cohort study included 317 patients with a prior cesarean delivery. Embryo transfer outcomes compared between the two groups based on the finding of a cesarean scar defect. The primary outcome was live birth rate, secondary outcomes of positive hCG, clinical intrauterine pregnancy, miscarriage rate. We fit a multivariate log binomial regression model with a priori variables for the primary and secondary outcomes, adjusting for patient and cycle characteristics.

Main results and the role of chance

There was no statistically significant difference in live birth rate among patients with a cesarean scar defect compared to those without, 28.0% vs 29.5% respectively (adjusted RR 0.95, 95% CI 0.68-1.35). There was no statistically significant difference in any of the secondary outcomes. Area of the defect and subgroup analysis of fresh vs frozen embryo transfer did not demonstrate any difference. Secondary analysis of patients with retroverted uterus had significantly lower live birth rates compared to those with an anteverted uterus, 17.7% vs 32.4%, respectively (adjusted RR 0.56, 95% CI 0.34-0.91).

Limitations, reasons for caution

Although a multivariate log binomial model was used to adjust for clinically relevant confounders, there remains the possibility for residual confounding given the observational nature of our study.

Wider implications of the findings

The results of our study suggest that, after adjusting for patient and cycle characteristics, the presence of a cesarean scar defect may not impact live birth outcomes and secondary analysis of the impact of uterine retroversion requires more study.

Trial registration number

Not applicable
An Adaptable Flying Fish Robotic Model for Aero- and Hydrodynamic Experimentation
Authors: Saro-Cortes Valeria , Cui Yuhe , Dufficy Tierney , Boctor Arsanious , Flammang Brooke E , Wissa Aimy W
Published: June 2022

Flying fishes (family Exocoetidae) are known for achieving multi-modal locomotion through air and water. Previous work on understanding this animal’s aerodynamic and hydrodynamic nature has been based on observations, numerical simulations, or experiments on preserved dead fish, and has focused primarily on flying pectoral fins. The first half of this paper details the design and validation of a modular flying fish inspired robotic model organism (RMO). The second half delves into a parametric aerodynamic study of flying fish pelvic fins, which to date have not been studied in-depth. Using wind tunnel experiments at a Reynolds number of 30,000, we investigated the effect of the pelvic fin geometric parameters on aerodynamic efficiency and longitudinal stability. The pelvic fin parameters investigated in this study include the pelvic fin pitch angle and its location along the body. Results show that the aerodynamic efficiency is maximized for pelvic fins located directly behind the pectoral fins and is higher for more positive pitch angles. In contrast, pitching stability is neither achievable for positive pitching angles nor pelvic fins located directly below the pectoral fin. Thus, there is a clear a trade-off between stability and lift generation, and an optimal pelvic fin configuration depends on the flying fish locomotion stage, be it gliding, taxiing, or taking off. The results garnered from the RMO experiments are insightful for understanding the physics principles governing flying fish locomotion and designing flying fish inspired aerial-aquatic vehicles.
Temperature Extreme Events Decrease Endocrine and Immune Reactive Scope in Bullfrogs (Lithobates catesbeianus)
Authors: Lima Alan Siqueira , de Figueredo Aymam Cobo , Floreste Felipe Rangel , Garcia Neto Patrício Getúlio , Gomes Fernando Ribeiro , Titon Stefanny Christie Monteiro
Published: June 2022

Currently, effects of increased atmospheric temperature, in the context of ongoing climate change, have been investigated in multiple organisms and levels of biological organization. While there has been a focus on the impacts of increased mean temperature, an emergent and equally important point is the consequences of recurrent exposure to extreme temperature events, simulating heat waves. This study investigated the effects of serial exposure to high temperatures on immune and endocrine variables before and after exposure to an acute secondary stressor in bullfrogs (Lithobates catesbeianus). Adult males were divided into three groups and subjected to three thermal regimes: control (c; constant 22°C); experimental 1 (E1; kept at 22°C and exposed to 4 days of 30°C every 16 days); and experimental 2 (E2; kept at 22°C and exposed to 4 days of 30°C every 6 days). Blood samples were collected on the last day of key extreme heat events. Two weeks after the last extreme heat event, animals were subjected to restraint stress (1 h) and sampled again. Blood samples were used to determine neutrophil: lymphocyte ratio, plasma bacterial killing ability, as well as, corticosterone and plasma testosterone levels. Overall, we found exposure to extreme heat events did not affect immune and endocrine variables over time. Meanwhile, the previous exposure to extreme heat events modulated the responsiveness to restraint. The amplitude of increased corticosterone plasma levels and neutrophil: lymphocyte ratio in response to restraint decreased with the number of previous exposures to extreme heat events. These results suggest that exposure to extreme climatic events has hidden effects on bullfrog's stress response, expressed as diminished reactive scope to a novel stressor. This represents a highly deleterious facet of climate change since diminished responsiveness prevents proper coping with wildlife challenges.
SARS-CoV-2 seroprevalence at urban and rural sites in Kaduna State, Nigeria, during October/November 2021, immediately prior to detection of the Omicron variant
Authors: Chechet Gloria D , Kwaga Jacob K P , Yahaya Joseph , Noyes Harry , MacLeod Annette , Adamson Walt E
Published: June 2022

Background

Nigeria is Africa’s most populated country. By November 2021 it had experienced three waves of SARS-CoV-2 infection. Peer-reviewed seroprevalence data assessing the proportion of the Nigerian population that have been infected were extremely limited.

Methods

We conducted a serosurvey in one urban site (n = 400) and one rural site (n = 402) in Kaduna State, Nigeria between 11 October 2021 and 8 November 2021. Z-tests were used to compare seroprevalence across age groups, locations and sexes. T tests were used to determine whether age or household size are associated with seropositivity. Associations between seropositivity and recent history of common Covid-19 symptoms were tested using logistic regression.

Results

SARS-CoV-2 antibodies were detected in 42.5% an 53.5% of participants at the urban and rural sites, respectively The overall age- and sex- stratified seroprevalence was 43.7% (42.2% for unvaccinated individuals). The data indicate an infection rate in Kaduna State ≥359-fold the rate derived from polymerase chain reaction-confirmed cases. In the urban site, seroprevalence among females and participants aged <20 was lower than other groups. Reporting loss of sense of taste and/or smell was strongly associated with seropositive status. Associations with seropositivity were also found for the reporting of dry cough, fever, headache, nausea and sore throat.

Conclusions

This study provides baseline SARS-CoV-2 seroprevalence in Kaduna State, Nigeria, immediately prior to the spread of the Omicron variant. It indicates that in October/November 2021, approximately 56% of the population did not have detectable antibodies, and population subgroups with particularly low seroprevalence remain. It highlights limitations in using PCR-confirmed cases to estimate infection rates. The data will inform public health strategies in Nigeria and other sub-Saharan African countries with limited SARS-CoV-2 seroprevalence data.
Enhancing research data infrastructure to address the opioid epidemic: the Opioid Overdose Network (O2-Net)
Authors: Lenert Leslie A , Zhu Vivienne , Jennings Lindsey , McCauley Jenna L , Obeid Jihad S , Ward Ralph , Hassanpour Saeed , Marsch Lisa A , Hogarth Michael , Shipman Perry , Harris Daniel R , Talbert Jeffery C
Published: June 2022

Opioid Overdose Network is an effort to generalize and adapt an existing research data network, the Accrual to Clinical Trials (ACT) Network, to support design of trials for survivors of opioid overdoses presenting to emergency departments (ED). Four institutions (Medical University of South Carolina [MUSC], Dartmouth Medical School [DMS], University of Kentucky [UK], and University of California San Diego [UCSD]) worked to adapt the ACT network. The approach that was taken to enhance the ACT network focused on 4 activities: cloning and extending the ACT infrastructure, developing an e-phenotype and corresponding registry, developing portable natural language processing tools to enhance data capture, and developing automated documentation templates to enhance extended data capture. Overall, initial results suggest that tailoring of existing multipurpose federated research networks to specific tasks is feasible; however, substantial efforts are required for coordination of the subnetwork and development of new tools for extension of available data. The initial output of the project was a new approach to decision support for the prescription of naloxone for home use in the ED, which is under further study within the network.
Urinary Excretion Profile of Cannabinoid Analytes Following Acute Administration of Oral and Vaporized Cannabis in Infrequent Cannabis Users
Authors: Sholler Dennis J , Zamarripa C Austin , Spindle Tory R , Martin Erin L , Kuntz David , Vandrey Ryan , Grabenauer Megan
Published: June 2022

Traditionally, smoking has been the predominant method for administering cannabis, but alternative routes of administration have become more prevalent. Additionally, research examining urinary cannabinoid excretion profiles has primarily focused on 11-nor-9-carboxy-∆9-tetrahydrocannabinol (∆9-THC-COOH), a metabolite of ∆9-tetrahydrocannabinol (∆9-THC), as the primary analyte. The aim of the current study was to characterize the urinary excretion profile of ∆9-THC-COOH, ∆9-THC, ∆8-tetrahydrocannabinol (∆8-THC), 11-hydroxy-∆9-tetrahydrocannabinol (11-OH-∆9-THC), ∆9-tetrahydrocannabivarin (THCV), 11-nor-∆9-tetrahydrocannabivarin-9-carboxlic acid (THCV-COOH), cannabidiol (CBD), cannabinol (CBN) and 8,11-dihydroxytetrahydrocannabinol (8,11-diOH-∆9-THC) following controlled administration of both oral and vaporized cannabis. Participants (n = 21, 11 men/10 women) who were infrequent cannabis users ingested cannabis-containing brownies (0, 10 and 25 mg ∆9-THC) and inhaled vaporized cannabis (0, 5 and 20 mg ∆9-THC) across six double-blind outpatient sessions. Urinary concentrations of ∆9-THC analytes were measured at baseline and for 8 h after cannabis administration. Sensitivity, specificity and agreement between the three immunoassays (IAs) for ∆9-THC-COOH (cutoffs of 20, 50 and 100 ng/mL) and liquid chromatography–tandem mass spectrometry (LC–MS-MS) analyses (confirmatory cutoff concentrations of 15 ng/mL) were assessed. Urinary concentrations for ∆9-THC-COOH, ∆9-THC, 11-OH-∆9-THC, THCV, CBN and 8,11-diOH-∆9-THC all peaked at 5–6 h and 4 h following oral and vaporized cannabis administration, respectively. At each active dose, median maximum concentrations (Cmax) for detected analytes were quantitatively higher after oral cannabis administration compared to vaporized. Using current recommended federal workplace drug-testing criteria (screening via IA with a cutoff of ≥50 ng/mL and confirmation via LC–MS-MS at a cutoff of ≥15 ng/mL), urine specimens tested positive for ∆9-THC-COOH in 97.6% of oral sessions and 59.5% of vaporized sessions with active ∆9-THC doses. These data indicate that while ∆9-THC-COOH may serve as the most consistent confirmatory analyte under the current drug-testing guidelines, future work examining 11-OH-∆9-THC under similar parameters could yield an alternative analyte that may be helpful in distinguishing between licit and illicit cannabis products.
Xylazine: Pharmacology Review and Prevalence and Drug Combinations in Forensic Toxicology Casework
Authors: Kacinko Sherri L , Mohr Amanda L A , Logan Barry K , Barbieri Edward J
Published: June 2022

Xylazine, an alpha-2 receptor agonist used in veterinary medicine for its sedative and muscle-relaxant effects, has been reported in forensic toxicology casework since the 1980s. It is not approved for human use, but it is used as an adulterant in heroin and illicit fentanyl. The prevalence and concentrations of xylazine in 2.5 years (January 2019–June 2021) of driving under the influence of drugs (DUID) and medico-legal death investigation (MDI) cases was investigated, including other drugs detected in combination with xylazine. Of over 170,000 cases screened for xylazine, 97% were classified as MDI. Over the course of the study period, the prevalence and geographical spread of xylazine increased. Overall, 2.8% of DUID and 2.1% of MDI cases screened positive for xylazine with concentrations of 5.1–450 ng/mL (mean = 36 ng/mL) and 5.0–11,000 ng/mL (mean = 41 ng/mL), respectively. Two MDI cases which had xylazine concentrations of 9,100 and 11,000 ng/mL were drug overdose suicides that did not involve any opioids. Opioids, primarily fentanyl and/or a fentanyl byproduct/metabolite were detected in 100% of DUID and all but two MDI cases. After opioids, stimulants, phyto-cannabinoids and benzodiazepines were the most common drug classes detected in conjunction with xylazine in both DUID and MDI casework. In summary, xylazine exposure continues to increase, mostly through the adulteration of illicit opioids. There is an extensive overlap in the concentrations between living and deceased individuals, making it difficult to interpret the role of the drug in MDI or DUID cases without other case information.