No association between LH levels and ovarian response in oocyte donors triggered with gonadotropin-releasing hormone agonist: A prospective study.

OBJECTIVE: Are circulating luteinizing hormone (LH) levels predictive of ovarian response in oocyte donors triggered with gonadotropin-releasing hormone (GnRH) agonists? STUDY DESIGN: A prospective cohort study with 224 oocyte donation cycles between 2021 and 2022 at a single center, examined the relationship between circulating luteinizing hormone (LH) levels and ovarian response. Oocyte donors underwent GnRH antagonist downregulation followed by GnRH agonist trigger. LH, estradiol, and progesterone levels were measured on day one of stimulation, trigger-day and 12 h post-trigger. Oocyte retrieval and maturity rates were analyzed using univariate and multivariate analyses, and the correlation between post-trigger LH levels and outcomes was assessed by Pearson's correlation test. A significance level of p < 0.05 was used. RESULTS: Mean age was 26 ± 4.3 years, mean body mass index (BMI, kg/m2) was 22.6 ± 3.2 and mean antral follicle count (AFC) was 21.7 ± 8.2. Post-trigger LH levels averaged 51.3 IU/L (SD 34.8), and oocyte retrieval rate and maturity rates were 112,7% (+/-48,1%) and 77,8% (+/- 17,2%), respectively. No significant differences were found in these outcomes for donors with post-trigger LH values below and above 15 IU/L (Mann Whitney's p > 0.05). However, exploratory analyses revealed that post-trigger LH values < 22 IU/L and basal LH levels < 4 IU/L were associated with significantly lower oocyte retrieval rate (90 % vs 110 %, p = 0.019 and 100 % vs 110 %, p = 0.019, respectively). CONCLUSIONS: This study, a first in exclusively focusing on oocyte donors, did not support the previously reported LH value of 15 IU/L as predictive of suboptimal ovarian response. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov Identifier: NCT05109403.

Implicit bias in diagnosing mosaicism amongst preimplantation genetic testing providers: results from a multicenter study of 36 395 blastocysts.

STUDY QUESTION: Does the diagnosis of mosaicism affect ploidy rates across different providers offering preimplantation genetic testing for aneuploidies (PGT-A)? SUMMARY ANSWER: Our analysis of 36 395 blastocyst biopsies across eight genetic testing laboratories revealed that euploidy rates were significantly higher in providers reporting low rates of mosaicism. WHAT IS KNOWN ALREADY: Diagnoses consistent with chromosomal mosaicism have emerged as a third category of possible embryo ploidy outcomes following PGT-A. However, in the era of mosaicism, embryo selection has become increasingly complex. Biological, technical, analytical, and clinical complexities in interpreting such results have led to substantial variability in mosaicism rates across PGT-A providers and clinics. Critically, it remains unknown whether these differences impact the number of euploid embryos available for transfer. Ultimately, this may significantly affect clinical outcomes, with important implications for PGT-A patients. STUDY DESIGN, SIZE, DURATION: In this international, multicenter cohort study, we reviewed 36 395 consecutive PGT-A results, obtained from 10 035 patients across 11 867 treatment cycles, conducted between October 2015 and October 2021. A total of 17 IVF centers, across eight PGT-A providers, five countries and three continents participated in the study. All blastocysts were tested using trophectoderm biopsy and next-generation sequencing. Both autologous and donation cycles were assessed. Cycles using preimplantation genetic testing for structural rearrangements were excluded from the analysis. PARTICIPANTS/MATERIALS, SETTING, METHODS: The PGT-A providers were randomly categorized (A to H). Providers B, C, D, E, F, G, and H all reported mosaicism, whereas Provider A reported embryos as either euploid or aneuploid. Ploidy rates were analyzed using multilevel mixed linear regression. Analyses were adjusted for maternal age, paternal age, oocyte source, number of embryos biopsied, day of biopsy, and PGT-A provider, as appropriate. We compared associations between genetic testing providers and PGT-A outcomes, including the number of chromosomally normal (euploid) embryos determined to be suitable for transfer. MAIN RESULTS AND THE ROLE OF CHANCE: The mean maternal age (±SD) across all providers was 36.2 (±5.2). Our findings reveal a strong association between PGT-A provider and the diagnosis of euploidy and mosaicism. Amongst the seven providers that reported mosaicism, the rates varied from 3.1% to 25.0%. After adjusting for confounders, we observed a significant difference in the likelihood of diagnosing mosaicism across providers (P < 0.001), ranging from 6.5% (95% CI: 5.2-7.4%) for Provider B to 35.6% (95% CI: 32.6-38.7%) for Provider E. Notably, adjusted euploidy rates were highest for providers that reported the lowest rates of mosaicism (Provider B: euploidy, 55.7% (95% CI: 54.1-57.4%), mosaicism, 6.5% (95% CI: 5.2-7.4%); Provider H: euploidy, 44.5% (95% CI: 43.6-45.4%), mosaicism, 9.9% (95% CI: 9.2-10.6%)); and Provider D: euploidy, 43.8% (95% CI: 39.2-48.4%), mosaicism, 11.0% (95% CI: 7.5-14.5%)). Moreover, the overall chance of having at least one euploid blastocyst available for transfer was significantly higher when mosaicism was not reported, when we compared Provider A to all other providers (OR = 1.30, 95% CI: 1.13-1.50). Differences in diagnosing and interpreting mosaic results across PGT-A laboratories raise further concerns regarding the accuracy and relevance of mosaicism predictions. While we confirmed equivalent clinical outcomes following the transfer of mosaic and euploid blastocysts, we found that a significant proportion of mosaic embryos are not used for IVF treatment. LIMITATIONS, REASONS FOR CAUTION: Due to the retrospective nature of the study, associations can be ascertained, however, causality cannot be established. Certain parameters such as blastocyst grade were not available in the dataset. Furthermore, certain platform-related and clinic-specific factors may not be readily quantifiable or explicitly captured in our dataset. As such, a full elucidation of all potential confounders accounting for variability may not be possible. WIDER IMPLICATIONS OF THE FINDINGS: Our findings highlight the strong need for standardization and quality assurance in the industry. The decision not to transfer mosaic embryos may ultimately reduce the chance of success of a PGT-A cycle by limiting the pool of available embryos. Until we can be certain that mosaic diagnoses accurately reflect biological variability, reporting mosaicism warrants utmost caution. A prudent approach is imperative, as it may determine the difference between success or failure for some patients. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the Torres Quevedo Grant, awarded to M.P. (PTQ2019-010494) by the Spanish State Research Agency, Ministry of Science and Innovation, Spain. M.P., L.B., A.R.L., A.L.R.d.C.L., N.P.P., M.P., D.S., F.A., A.P., B.M., L.D., F.V.M., D.S., M.R., E.P.d.l.B., A.R., and R.V. have no competing interests to declare. B.L., R.M., and J.A.O. are full time employees of IB Biotech, the genetics company of the Instituto Bernabeu group, which performs preimplantation genetic testing. M.G. is a full time employee of Novagen, the genetics company of Cegyr, which performs preimplantation genetic testing. TRIAL REGISTRATION NUMBER: N/A.

Effectiveness and Safety of Teriflunomide in Relapsing-Remitting Multiple Sclerosis and Improvements in Quality of Life: Results from the Real-World TERICARE Study.

INTRODUCTION: Teriflunomide is a once-daily oral immunomodulator approved for relapsing forms of multiple sclerosis (MS) or relapsing-remitting multiple sclerosis (RRMS; depending on the local label), based on extensive evidence from clinical trials and a real-world setting on efficacy, tolerability and patient-reported benefits. The TERICARE study assessed the impact of teriflunomide treatment over 2 years on health-related quality of life (HRQoL) and some of the most common and disabling symptoms of MS, such as fatigue and depression. METHODS: This prospective observational study in Spain included RRMS patients treated with teriflunomide for ≤ 4 weeks. The following patient-reported outcomes (PROs) were collected at baseline and every 6 months for 2 years: the 29-item Multiple Sclerosis Impact Scale version 2 (MSIS-29), the 21-item Modified Fatigue Impact Scale (MFIS-21), the Beck Depression Inventory (BDI-II), the Short Form (SF)-Qualiveen and the Treatment Satisfaction Questionnaire for Medication v1.4 (TSQM). Annualised relapse rate (ARR), disability progression according to the Expanded Disability Status Scale (EDSS), and no evidence of disease activity (NEDA-3) were also assessed. RESULTS: A total of 325 patients were analysed. Patients had a mean (SD) age of 43.2 years (10.4), a mean baseline EDSS score of 1.75 (1.5), a mean number of relapses in the past 2 years of 1.5 (0.7), and 64% had received prior disease-modifying therapy (DMT). Patients showed significant improvements in the psychological domain of MSIS-29 from 35.9 (26.6) at baseline to 29.4 (25.5) at 18 months (p = 0.004) and 29.0 (24.6) at 24 months (p = 0.002). Levels of fatigue and depression were also reduced. After 2 years of treatment with teriflunomide, ARR was reduced to 0.17 (95% CI 0.14-0.21) from the baseline of 0.42 (95% CI 0.38-0.48), representing a 60.1% reduction. Mean EDSS scores remained stable during the study, and 79.9% of patients showed no disability progression. 54.7% of patients achieved NEDA-3 in the first 12 months, which increased to 61.4% during months 12-24. Patients reported increased satisfaction with treatment over the course of the study, regardless of whether they were DMT naive or not. CONCLUSION: Teriflunomide improves psychological aspects of HRQoL and maintains low levels of fatigue and depression. Treatment with teriflunomide over 2 years is effective in reducing ARR and disability progression.

The genomic history of the indigenous people of the Canary Islands.

The indigenous population of the Canary Islands, which colonized the archipelago around the 3rd century CE, provides both a window into the past of North Africa and a unique model to explore the effects of insularity. We generate genome-wide data from 40 individuals from the seven islands, dated between the 3rd-16rd centuries CE. Along with components already present in Moroccan Neolithic populations, the Canarian natives show signatures related to Bronze Age expansions in Eurasia and trans-Saharan migrations. The lack of gene flow between islands and constant or decreasing effective population sizes suggest that populations were isolated. While some island populations maintained relatively high genetic diversity, with the only detected bottleneck coinciding with the colonization time, other islands with fewer natural resources show the effects of insularity and isolation. Finally, consistent genetic differentiation between eastern and western islands points to a more complex colonization process than previously thought.

Long-term storage of vitrified oocytes does not affect pregnancy and live birth rates: analysis of 5362 oocyte donation cycles.

RESEARCH QUESTION: Does long-term storage of vitrified oocytes affect laboratory and reproductive outcomes after intracytoplasmic sperm injection? DESIGN: Retrospective cohort study including 41,783 vitrified-warmed oocytes from 5362 oocyte donation cycles between 2013 and 2021. Five categories of storage time were established to analyse its effect on clinical and reproductive outcomes (≤1 year [reference group], 1-2 years, 2-3 years, 3-4 years and >4 years). RESULTS: The mean number of warmed oocytes was 8.0 ± 2.5 oocytes. Oocyte storage time ranged from 3 days to 8.2 years (mean: 0.7 ± 0.9). Mean oocyte survival (90.2% ± 14.7% overall) did not significantly decrease with longer storage time after adjusting for confounders (88.9% for time >4 years, P = 0.963). A linear regression model did not show a significant effect of oocyte storage time on fertilization rate (about 70% in all time categories) (P > 0.05). Reproductive outcomes after the first embryo transfer were statistically comparable across storage times (P > 0.05 for all categories). Longer term oocyte storage (>4 years) did not affect the chances of clinical pregnancy (OR 0.700, 95% CI 0.423 to 1.158, P = 0.2214) or live birth (OR 0.716, 95% CI 0.425 to 1.208, P = 0.2670). CONCLUSIONS: Oocyte survival, fertilization rate, pregnancy and live birth rates are not affected by the time spent by vitrified oocytes in vapour-phase nitrogen tanks.

Exonic genetic variants associated with unexpected fertilization failure and zygotic arrest after ICSI: a systematic review.

Fertilization failure (FF) and zygotic arrest after ICSI have a huge effect on both patients and clinicians, but both problems are usually unexpected and cannot be properly diagnosed. Fortunately, in recent years, gene sequencing has allowed the identification of multiple genetic variants underlying failed ICSI outcomes, but the use of this approach is still far from routine in the fertility clinic. In this systematic review, the genetic variants associated with FF, abnormal fertilization and/or zygotic arrest after ICSI are compiled and analyzed. Forty-seven studies were included. Data from 141 patients carrying 121 genetic variants affecting 16 genes were recorded and analyzed. In total, 27 variants in PLCZ1 (in 50 men) and 26 variants in WEE2 (in 24 women) are two of the factors related to oocyte activation failure that could explain a high percentage of male-related and female-related FF. Additional variants identified were reported in WBP2NL, ACTL9, ACTLA7, and DNAH17 (in men), and TUBB8, PATL2, TLE6, PADI6, TRIP13, BGT4, NLRP5, NLRP7, CDC20 and ZAR1 (in women). Most of these variants are pathogenic or potentially pathogenic (89/121, 72.9%), as demonstrated by experimental and/or in silico approaches. Most individuals carried bi-allelic variants (89/141, 63.1%), but pathogenic variants in heterozygosity have been identified for PLCZ1 and TUBB8. Clinical treatment options for affected individuals, such as chemical-assisted oocyte activation (AOA) or PLCZ1 cRNA injection in the oocyte, are still experimental. In conclusion, a genetic study of known pathogenic variants may help in diagnosing recurrent FF and zygotic arrest and guide patient counselling and future research perspectives.

Design of a Nutritional Survey to Detect High Dietary Salt Intakes and Its Usefulness in Primary Care Compared to 24-Hour Urine Sodium Determination.

Many population studies report salt intakes that exceed the WHO recommendation (2 g/day of Na+ or 5 g/day of salt). We do not have tools for detecting high salt intakes that are easy to apply in primary health care (PHC). We propose the development of a survey to screen for high salt intake in PHC patients. A cross-sectional study of 176 patients determines the responsible foods, and a study of 61 patients studies the optimal cut-off point and discriminant ability (ROC curve). We assessed the salt intake using a food frequency questionnaire and a 24 h dietary recall and used a factor analysis to identify the foods with the highest contribution to be included in a high intake screening questionnaire. We used 24 h urinary sodium as a gold standard. We identified 38 foods and 14 factors representing a high intake, explaining a significant proportion of the total variance (50.3%). Significant correlations (r > 0.4) were obtained between nutritional survey scores and urinary sodium excretion, allowing us to detect patients who exceed salt intake recommendations. For sodium excretion ≥ 2.4 g/day, the survey has a sensitivity of 91.4%, a specificity of 96.2% and an area under the curve of 0.94. For a prevalence of high consumption of 57.4%, the positive predictive value (PPV) was 96.9% and the negative predictive value (NPV) was 89.2%. We developed a screening survey for subjects with a high probability of high salt intake in primary health care, which could contribute to the reduction in diseases associated with this consumption.

Correlaciones entre la ingesta de sal en la dieta y la eliminación de sodio en la orina de 24 h en una muestra de población urbana de Cáceres / Correlations between dietary salt intake and 24-h urine sodium excretion in a sample of the urban population of Cáceres

Objetivo: La ingesta excesiva de sal se asocia a mayores cifras de tensión arterial y prevalencia de enfermedades cardiovasculares. La OMS recomienda consumir menos de 5 g/día de sal (equivalente a 2 g de Na+/día). Identificar alimentos y comportamientos con mayor contribución al exceso de aporte facilitaría el consejo dietético preventivo. Diseño: Estudio observacional. Sitio: Centro de Salud Urbano en Cáceres. Medidas principales: Estimamos el consumo de sal mediante dos cuestionarios de frecuencia de consumo de alimentos (CFA), uno genérico y otro con alimentos con elevado contenido en Na+, y encuesta recordatoria de 24 h. Empleamos el programa Evalfinut para la valoración nutricional de la dieta. Analizamos correlaciones entre ingesta estimada de sal y eliminación de sodio en orina de 24 h (gold standard). Resultados: El 92% de la muestra presenta consumos superiores a las recomendaciones con ingestas equivalentes a 9,5 g/día de sal (3,7 g/día de Na+). Cuando la ingesta de sodio se determina por el cociente Na+/K+, el 79,54% tiene ingestas elevadas. La eliminación de sodio sigue una tendencia ligeramente creciente con el IMC. La percepción de consumo de sal es baja, el 56,3% lo considera «adecuado»; el 32,4% añade sal a los alimentos una vez servidos. Los CFA infraestiman la ingesta de Na+ y encuestas dirigidas aportan valores más elevados. La correlación entre CFA y eliminación urinaria de Na+ es débil. Conclusión: Debemos reducir la ingesta de sal aumentando la percepción del consumo, mejorando el conocimiento sobre el contenido de sal en alimentos e identificando a pacientes diana del consejo sanitario.(AU)

Objective: Excessive salt intake is associated with higher levels of AHT and prevalence of cardiovascular diseases. WHO recommends consuming less than 5 g/day of salt (equivalent to 2 g Na+/day). Identifying foods and behaviours with greater contribution to excess intake would facilitate preventive dietary advice. Design: Observational study. Site: Urban Health Center in Cáceres. Main measurements: We estimated salt consumption using two food consumption frequency (FFQ) questionnaires, one generic and one with high Na+ content, and a 24 h follow-up survey. We use the Evalfinut program for nutritional evaluation of the diet. We analyzed correlations between estimated salt intake and 24-h urine sodium elimination (gold standard). Results: 92% of the population had consumption higher than the recommendations with intakes equivalent to 9.5 g/day of salt (3.7 g/d of Na+). When sodium intake is determined by the Na+/K+ ratio, 79.54% have high intakes. Sodium elimination follows a slightly increasing trend with BMI. The perception of salt consumption is low, 56.3% consider it “adequate”. 32.4% add salt to food once served. FFQ underestimate Na+ intake and targeted surveys provide higher values. The correlation between FFQ and Na+ urinary elimination is weak. Conclusion: We must reduce salt intake by increasing the perception of consumption, improving knowledge about the salt content in food and identifying target patients of the health council.(AU)

[Correlations between dietary salt intake and 24-h urine sodium excretion in a sample of the urban population of Cáceres]. / Correlaciones entre la ingesta de sal en la dieta y la eliminación de sodio en la orina de 24 h en una muestra de población urbana de Cáceres.

OBJECTIVE: Excessive salt intake is associated with higher levels of AHT and prevalence of cardiovascular diseases. WHO recommends consuming less than 5 g/day of salt (equivalent to 2 g Na+/day). Identifying foods and behaviours with greater contribution to excess intake would facilitate preventive dietary advice. DESIGN: Observational study. SITE: Urban Health Center in Cáceres. MAIN MEASUREMENTS: We estimated salt consumption using two food consumption frequency (FFQ) questionnaires, one generic and one with high Na+ content, and a 24 h follow-up survey. We use the Evalfinut program for nutritional evaluation of the diet. We analyzed correlations between estimated salt intake and 24-h urine sodium elimination (gold standard). RESULTS: 92% of the population had consumption higher than the recommendations with intakes equivalent to 9.5 g/day of salt (3.7 g/d of Na+). When sodium intake is determined by the Na+/K+ ratio, 79.54% have high intakes. Sodium elimination follows a slightly increasing trend with BMI. The perception of salt consumption is low, 56.3% consider it "adequate". 32.4% add salt to food once served. FFQ underestimate Na+ intake and targeted surveys provide higher values. The correlation between FFQ and Na+ urinary elimination is weak. CONCLUSION: We must reduce salt intake by increasing the perception of consumption, improving knowledge about the salt content in food and identifying target patients of the health council.

Cáncer infantil: lo que debemos saber / Childhood cáncer: What we should know

Introducción: el cáncer representa una de las principales causas de muerte en niños, niñas y adolescentes a nivel global, el cual es superado escasamente por los accidentes. Las neoplasias más frecuentes son, en primer lugar, las leucemias; segundo, los tumores del Sistema Nervioso Central; y, tercero, los linfomas. Es importante destacar que no existe prevención, sin embargo, hay signos de alarma que si se logran identificar a tiempo pueden verse traducidos en una enfermedad más manejable con una sobrevida más alta y un mejor pronóstico. Objetivo: realizar una actualización académica sobre la importancia de los signos y síntomas de alarma del cáncer infantil. Método: se realizó una investigación documental y se utilizó como buscador de información científica Google Académico. Se evaluaron artículos de investigación y revisión de diferentes bases de datos, como: site de la Organización Mundial de la Salud, site de la Oficina Panamericana de la Salud, Revista de Pediatría Integral, American Cáncer Society y publicaciones periodísticas locales, entre otros. Conclusiones: el cáncer en la infancia y adolescencia no puede prevenirse; sin embargo, si los médicos de los distintos niveles de atención en salud pueden reconocer los signos de alarma ante un probable caso de malignidad, se lograrán referimientos oportunos, diagnósticos tempranos y una mejor tasa de curabilidad y sobrevida en los niños.

Introduction: Cancer represents one of the main causes of death in children and adolescents globally, which is barely surpassed by accidents. The most frequent neoplasms are leukemias first, Central Nervous System tumors second, and lymphomas third. It is important to note that there is no prevention, however, there are warning signs that, if identified in time, can be translated into a more manageable disease with higher survival and a better prognosis. Objective: To carry out an academic update on the importance of warning signs and symptoms of childhood cancer. Method: A documentary research was carried out and Google Scholar was used as a search engine for scientific information. Research and review articles from different databases were evaluated, such as: the World Health Organization site, the Pan American Health Office site, the Comprehensive Pediatrics Review, the American Cancer Society, local journalistic publications, among others. Conclusions: Cancer in childhood and adolescence cannot be prevented, however, if doctors at the different levels of health care can recognize the warning signs in the event of a probable case of malignancy, timely referrals, early diagnoses and a better curability rate will be achieved and survival in children.

Utility of Lymphocyte Transformation Test for Assisting Updated Roussel Uclaf Causality Assessment Method in Drug-Induced Liver Injury: A Case-Control Study.

Background: The Roussel Uclaf Causality Assessment Method (RUCAM) is a validated tool for assessing causality in cases of suspected drug-induced liver injury (DILI). However, RUCAM cannot discriminate between concomitant hepatotoxic drugs with the same temporal sequence. Objective: To analyse the utility of the lymphocyte transformation test (LTT) for assisting updated RUCAM in 45 patients and 40 controls with a clinical diagnosis of DILI. Methods: Suspected DILI cases were detected through the Prospective Pharmacovigilance Program from Laboratory Signals in Hospital (PPLSH) or by consultations. The controls completed the drug therapy with no adverse reactions during the study period. A receiver operating characteristics (ROC) curve analysis was performed to calculate the optimal cut-off value for the stimulation index (SI), corresponding to the largest sum for the specificity and sensitivity values of LTT for true DILI cases. Results: Out of 45 patients diagnosed with DILI, 42 cases were detected by the PPLSH, two cases by consultation and one case by both methods. Most DILI cases (64.4%) arose during hospitalization. According to the biochemical parameters, 24 cases (53.3%) had the hepatocellular phenotype, 14 (31.1%) had the cholestatic phenotype, and 7 cases (15.6%) had the mixed phenotype. Considering the severity criteria, 7 (15.5%) cases were classified as moderate DILI, and 4 (8.9%) were severe DILI; there were no fatal cases. A total of 149 drugs (median/case, 3; IQR, 2-5) were suspected to be involved in the DILI cases (RUCAM score ≥3). In 8 cases, only one drug was suspected, and polypharmacy (≥5 drugs) was identified in 29% of the cases. Of all DILI cases, 46 (30.9%) of the 149 suspected drugs produced positive LTT results, and the LTT was positive in 34 (75.5%) of the 45 patients. No exposed controls produced positive LTT results. The optimal cut-off of 1.95 for the SI was obtained with a sensitivity of 77% and specificity of 100% (area under the curve, 0.91; 95% asymptotic confidence interval 0.84-0.97; p < 0.001). The sensitivity of the hepatocellular phenotype was 92%. Conclusion: Our results demonstrate that LTT is an add on strengthening causality in cases of suspected idiosyncratic DILI, especially for patients with several suspected drugs and a hepatocellular phenotype.

How long can the sperm wait? Effect of incubation time on ICSI outcomes.

In sperm processing for IVF/ICSI incubation times differ considerably both between and within assisted reproduction facilities. There is no established consensus on the optimal sperm incubation timings to maximize pregnancy rates, and the few studies addressing this association rely on manual and operator-dependent methods for time recording. The present retrospective cohort study includes 1169 ICSI cycles using fresh semen processed by swim-up. An operator-independent, radiofrequency-based system was used to record sperm incubation times: from sample collection to swim-up (T1, 0.35 ± 0.26); from swim-up to ICSI (T2, 3.30 ± 2.2); and total time from sample collection to ICSI (T, 3.66 ± 2.26). In oocyte donation cycles, we observed a significant negative effect of T1 on fertilization rate (FR; generalized linear modelling regression, coeff. -0.20, p = 0.001); however, after analysing all times by deciles and by adjusted logistic regression, none of the time intervals had a significant effect on pregnancy (biochemical, clinical, and ongoing) and live birth (LB) rates (p > 0.05 for all outcomes). In cycles using the patient's oocytes, we observed a negative effect of T2 (ordinal regression, coeff. -0.25, p = 0.011) and T (-0.33, p = 0.005) on the mean morphological score of the embryo cohort. In these cycles, a trend associating longer values of T with higher LB rates was identified (OR = 1.47, p = 0.050), although this difference is likely not clinically significant. In conclusion, while longer sperm incubation in vitro may impact slightly both FRs and embryo morphology after ICSI, no adverse effects were detected on the reproductive outcomes.

Characterisation of Drug-Induced Liver Injury in Patients with COVID-19 Detected by a Proactive Pharmacovigilance Program from Laboratory Signals.

Coronavirus disease 2019 (COVID-19) has a wide spectrum of clinical manifestations. An elevation of liver damage markers has been observed in numerous cases, which could be related to the empirical use of potentially hepatotoxic drugs. The aim of this study was to describe the clinical and analytical characteristics and perform a causality analysis from laboratory signals available of drug-induced liver injury (DILI) detected by a proactive pharmacovigilance program in patients hospitalised for COVID-19 at La Paz University Hospital in Madrid (Spain) from 1 March 2020 to 31 December 2020. The updated Roussel Uclaf Causality Assessment Method (RUCAM) was employed to assess DILI causality. A lymphocyte transformation test (LTT) was performed on 10 patients. Ultimately, 160 patients were included. The incidence of DILI (alanine aminotransferase >5, upper limit of normal) was 4.9%; of these, 60% had previous COVID-19 hepatitis, the stay was 8.1 days longer and 98.1% were being treated with more than 5 drugs. The most frequent mechanism was hepatocellular (57.5%), with mild severity (87.5%) and subsequent recovery (88.1%). The most commonly associated drugs were hydroxychloroquine, azithromycin, tocilizumab and ceftriaxone. The highest incidence rate of DILI per 10,000 defined daily doses (DDD) was with remdesivir (992.7/10,000 DDD). Some 80% of the LTTs performed were positive, with a RUCAM score of ≥4. The presence of DILI after COVID-19 was associated with longer hospital stays. An immune mechanism has been demonstrated in a small subset of DILI cases.

Human oocyte meiotic maturation is associated with a specific profile of alternatively spliced transcript isoforms.

The transition from a transcriptionally active state (GV) to a transcriptionally inactive state (mature MII oocytes) is required for the acquisition of oocyte developmental competence. We hypothesize that the expression of specific genes at the in vivo matured (MII) stage could be modulated by posttranscriptional mechanisms, particularly regulation of alternative splicing (AS). In this study, we examined the transcriptional activity of GV oocytes after ovarian stimulation followed by oocyte pick-up and the landscape of alternatively spliced isoforms in human MII oocytes. Individual oocytes were processed and analyzed for transcriptional activity (GV), gene expression (GV and MII), and AS signatures (GV and MII) on HTA 2.0 microarrays. Samples were grouped according to maturation stage, and then subgrouped according to women's age and antral follicular count (AFC); array results were validated by quantitative polymerase chain reaction. Differentially expressed genes between GV and MII oocytes clustered mainly in biological processes related to mitochondrial metabolism. Interestingly, 16 genes that were related to the regulation of transcription and mitochondrial translation showed differences in alternatively spliced isoform profiles despite not being differentially expressed between groups. Altogether, our results contribute to our understanding of the role of AS in oocyte developmental competence acquisition.

Reproductive Outcomes in Lesbian Couples Undergoing Reception of Oocytes from Partner Versus Autologous In Vitro Fertilization/Intracytoplasmic Sperm Injection.

Purpose: This study aimed to compare reproductive outcomes after Reception of Oocytes from Partner (ROPA; also called reciprocal in vitro fertilization) with those after in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) with autologous oocytes, in lesbian couples. Methods: This was a retrospective matched cohort study of couples performing a first cycle of either ROPA (n = 60) or autologous IVF/ICSI (n = 120) between February 2012 and May 2018. Couples were matched 1:2 by age of the oocyte provider, day of embryo transfer (ET), and number of embryos transferred. Pregnancy and live birth rates after the first ET and cumulative results after all subsequent ETs performed until June 2019 were evaluated. Results: Reproductive outcomes were significantly better after ROPA at first ET: biochemical pregnancy 70.0% versus 47.5% (p = 0.004), clinical pregnancy 60.0% versus 40.0% (p = 0.011), ongoing pregnancy 60.0% versus 36.7% (p = 0.003), and live birth 57.1% versus 29.8% (p = 0.001). After adjusting for age, body mass index, and number of mature oocytes, we still observed a significant improvement across all outcomes in ROPA (live birth rate odds ratio [OR]: 3.05, 95% confidence interval [CI] 1.42-6.57). Cumulative pregnancy and live birth rates were also higher after ROPA (live birth 66.1% vs. 43.4% [p = 0.005]). The adjusted analysis result for cumulative live birth was OR: 2.51, 95% CI: 1.14-5.54. Conclusion: When medically indicated, ROPA can potentially improve reproductive outcomes for lesbian couples through the possibility of selecting the best combination between two oocyte providers and two gestational mothers, provided that both women wish to participate in the pregnancy plan.

Altered mitochondrial function in spermatozoa from patients with repetitive fertilization failure after ICSI revealed by proteomics.

BACKGROUND: Unexplained fertilization failure (FF), occurring in 1-3% of intracytoplasmic sperm injection (ICSI) cycles, results in both psychological and financial burden for the patients. However, the molecular causes behind FF remain largely unknown. Mass spectrometry is a powerful technique to identify and quantify proteins across samples; however, no study so far has used it to dissect the proteomic signature of spermatozoa with FF after ICSI. OBJECTIVE: To investigate whether sperm samples from patients suffering repetitive FF after ICSI display alterations in their protein content. MATERIAL AND METHODS: Seventeen infertile men were included: 5 patients presented FF in ≥3 consecutive ICSI cycles, while 12 patients had a fertilization rate >75% (controls). Individual sperm samples were subjected to 2D-LC-MS/MS. Both conventional and novel statistical approaches were used to identify differentially abundant proteins. Additionally, analysis of mitochondrial and proteasomal abundance and activity were performed, using Western blot, FACS analysis of JC-1 staining and AMC-peptide fluorometric assay. RESULTS: Four proteins presented lower abundance (FMR1NB, FAM209B, RAB2B, and PSMA1) in the FF group compared to controls, while five mitochondrial proteins presented higher abundance in FF (DLAT, ATP5H, SLC25A3, SLC25A6, and FH) (p < 0.05). The altered abundance of mitochondrial DLAT and proteasomal PSMA1 was corroborated by Western blot. Of relevance, novel stable-protein pair analysis identified 73 correlations comprising 28 proteins within controls, while different mitochondrial proteins (ie, PDHA2, PHB2, and ATP5F1D) lost >50% of these correlations in specific FF samples pointing out specific mitochondrial deregulations. DISCUSSION: This is the first proteomic analysis of spermatozoa from patients who resulted in fertilization failure after ICSI. The altered proteins, most of them related to mitochondrial function, could help to identify diagnostic/prognostic markers of fertilization failure and could further dissect the molecular paternal contribution to reach successful fertilization. CONCLUSION: Sperm samples from patients with FF after ICSI present altered abundance of different proteins, including mainly mitochondrial proteins.

Trophoblast attachment to the endometrial epithelium elicits compartment-specific transcriptional waves in an in-vitro model.

RESEARCH QUESTION: Which are the early compartment-specific transcriptional responses of the trophoblast and the endometrial epithelium throughout early attachment during implantation? DESIGN: An endometrial epithelium proxy (cell line Ishikawa) was co-cultured with spheroids of a green fluorescent protein (GFP) expressing trophoblast cell line (JEG-3). After 0, 8 and 24 h of co-culture, the compartments were sorted by fluorescence-activated cell sorting; GFP+ (trophoblast), GFP- (epithelium) and non-co-cultured control populations were analysed (in triplicate) by RNA-seq and gene set enrichment analysis (GSEA). RESULTS: Trophoblast challenge induced a wave of transcriptional changes in the epithelium that resulted in 295 differentially regulated genes involving epithelial to mesenchymal transition (EMT), cell movement, apoptosis, hypoxia, inflammation, allograft rejection, myogenesis and cell signalling at 8 h. Interestingly, many of the enriched pathways were subsequently de-enriched by 24 h (i.e. EMT, cell movement, allograft rejection, myogenesis and cell signalling). In the trophoblast, the co-culture induced more transcriptional changes and regulation of a variety of pathways. A total of 1247 and 481 genes were differentially expressed after 8 h and from 8 to 24 h, respectively. Angiogenesis and hypoxia were over-represented at both stages, while EMT and cell signalling only were at 8 h; from 8 to 24 h, inflammation and oestrogen response were enriched, while proliferation was under-represented. CONCLUSIONS: Successful attachment produced a series of dynamic changes in gene expression, characterized by an overall early and transient transcriptional up-regulation in the receptive epithelium, in contrast to a more dynamic transcriptional response in the trophoblast.

Incidence of Suspected Serious Adverse Drug Reactions in Corona Virus Disease-19 Patients Detected by a Pharmacovigilance Program by Laboratory Signals in a Tertiary Hospital in Spain: Cautionary Data.

BACKGROUND: From March to April 2020, Spain was the center of the SARS-CoV-2 pandemic, particularly Madrid with approximately 30% of the cases in Spain. The aim of this study is to report the suspected serious adverse drug reactions (SADRs) in COVID-19 patients vs. non-COVID-19 patients detected by the prospective pharmacovigilance program based on automatic laboratory signals (ALSs) in the hospital (PPLSH) during that period. We also compared the results with the suspected SADRs detected during the same period for 2019. METHODS: All ALSs that reflected potential SADRs including neutropenia, pancytopenia, thrombocytopenia, anemia, eosinophilia, leukocytes in cerebrospinal fluid, hepatitis, pancreatitis, acute kidney injury, rhabdomyolysis, and hyponatremia were prospectively monitored in hospitalized patients during the study periods. We analyzed the incidence and the distribution of causative drugs for the COVID-19 patients. RESULTS: The incidence rate of SADRs detected in the COVID-19 patients was 760.63 (95% CI 707.89-816.01) per 10,000 patients, 4.75-fold higher than the SADR rate for non-COVID-19 patients (160.15 per 10,000 patients, 95% CI 137.09-186.80), and 5.84-fold higher than the SADR rate detected for the same period in 2019 (130.19 per 10,000 patients, 95% CI 109.53-154.36). The most frequently related drugs were tocilizumab (59.84%), dexketoprofen (13.93%), azithromycin (8.43%), lopinavir-ritonavir (7.35%), dexamethasone (7.62%), and chloroquine/hydroxychloroquine (6.91%). CONCLUSIONS: The incidence rate of SADRs detected by the PPSLH in patients with COVID-19 was 4.75-fold higher than that of the non-COVID-19 patients. Caution is recommended when using medications for COVID-19 patients, especially drugs that are hepatotoxic, myotoxic, and those that induce thromboembolic events.

Identification of research priorities in infertility and assisted reproduction: an international, multicentre study.

RESEARCH QUESTION: What are the main research interests among patients of assisted reproductive technologies (ART)? DESIGN: Cross-sectional study consisting of an anonymous online survey sent to 2112 patients from eight centres in four countries in 2018. Patients were asked to identify research questions relevant to them in the field of infertility and ART. Answers were categorized into topics and ranked by frequency. A long list of the top 30 research topics was extracted from the aggregate results, from which a short list of the top 10 research topics was created. Ten research questions were finally formulated. RESULTS: A total of 945 responses were analysed. Main interests were side-effects, success rates, infertility prevention and emotional support. The 10 research questions were: 1. What are the side-effects of drugs used in ART treatments? (51.6%). 2. What are the most effective methods to cope with infertility from the psychological point of view? (35.7%). 3. What effects could diet have on fertility? (25.9%). 4. What are ART success rates per clinical profile? (24.8%). 5. Are there some habits and lifestyle factors that could prevent infertility? (20.0%). 6. What are the long-term risks associated with ART in mother and child? (18.5%). 7. Are alternative therapies such as acupuncture, yoga and meditation effective to treat/prevent infertility? (18.5%). 8. What is the impact of exercise on fertility? (15.4%). 9. How does oocyte quantity and quality affect fertility? (9.5%). 10. What are the genetic patterns or hereditary conditions causing/related to infertility? (9.5%). CONCLUSIONS: Researchers and clinicians should keep in mind that, in addition to success rates and safety, patients greatly value research into causes, prevention and emotional aspects of infertility.