Common genetic variation in KATNAL1 non-coding regions is involved in the susceptibility to severe phenotypes of male infertility.

BACKGROUND: Previous studies in animal models evidenced that genetic mutations of KATNAL1, resulting in dysfunction of its encoded protein, lead to male infertility through disruption of microtubule remodelling and premature germ cell exfoliation. Subsequent studies in humans also suggested a possible role of KATNAL1 single-nucleotide polymorphisms in the development of male infertility as a consequence of severe spermatogenic failure. OBJECTIVES: The main objective of the present study is to evaluate the effect of the common genetic variation of KATNAL1 in a large and phenotypically well-characterised cohort of infertile men because of severe spermatogenic failure. MATERIALS AND METHODS: A total of 715 infertile men because of severe spermatogenic failure, including 210 severe oligospermia and 505 non-obstructive azoospermia patients, as well as 1058 unaffected controls were genotyped for three KATNAL1 single-nucleotide polymorphism taggers (rs2077011, rs7338931 and rs2149971). Case-control association analyses by logistic regression assuming different models and in silico functional characterisation of risk variants were conducted. RESULTS: Genetic associations were observed between the three analysed taggers and different severe spermatogenic failure groups. However, in all cases, the haplotype model (rs2077011*C | rs7338931*T | rs2149971*A) better explained the observed associations than the three risk alleles independently. This haplotype was associated with non-obstructive azoospermia (adjusted p = 4.96E-02, odds ratio = 2.97), Sertoli-cell only syndrome (adjusted p = 2.83E-02, odds ratio = 5.16) and testicular sperm extraction unsuccessful outcomes (adjusted p = 8.99E-04, odds ratio = 6.13). The in silico analyses indicated that the effect on severe spermatogenic failure predisposition could be because of an alteration of the KATNAL1 splicing pattern. CONCLUSIONS: Specific allelic combinations of KATNAL1 genetic polymorphisms may confer a risk of developing severe male infertility phenotypes by favouring the overrepresentation of a short non-functional transcript isoform in the testis.

Analysing endometrial microbiome: methodological considerations and recommendations for good practice.

There is growing evidence that the upper female genital tract is not sterile, harbouring its own microbial communities. However, the significance and the potential effect of endometrial microorganisms on reproductive functions remain to be fully elucidated. Analysing the endometrial microbiome, the microbes and their genetic material present in the endometrium, is an emerging area of study. The initial studies suggest it is associated with poor reproductive outcomes and with different gynaecological pathologies. Nevertheless, studying a low-biomass microbial niche as is endometrium, the challenge is to conduct well-designed and well-controlled experiments in order to avoid and adjust for the risk of contamination, especially from the lower genital tract. Herein, we aim to highlight methodological considerations and propose good practice recommendations for future endometrial microbiome studies.

Effectiveness of a psychological intervention focused on stress management for women prior to IVF.

Objective: To evaluate the effectiveness of a psychological intervention focused on stress management in women who are candidates for in vitro fertilisation (IVF).Method: Pre-post study with two groups (n = 26). The psychological intervention group (IG) received a 90-minute session that included psychoeducation, relaxation training and coping skills. The control group (CG) did not receive any psychological intervention. All participants later underwent IVF. In the initial and final evaluations, state anxiety, emotional imbalance, adaptive resources and quality of life were measured. In the initial evaluation, demographic and clinical variables were also measured.Results: In the IG, the final evaluation reflected: (1) decreased levels of anxiety and emotional imbalance; (2) the perception of enhanced quality of life. Moreover, among the participants who received the psychological intervention and later achieved a successful IVF, the level of anxiety appeared to have decreased more strongly.Conclusions: A brief intervention focused on stress management can benefit the psychological adjustment of women who are candidates for IVF, reducing the anxiety they may experience in this regard. However, our conclusions are based on a small sample, and so should be considered with caution. Nevertheless, these results are promising and highlight the advisability of further research.

Application of a sperm survival test: Results from an external quality control programme.

OBJECTIVE: The study aim is to determine which type of material - pipette tips or culture medium - is more appropriate for use in a cytotoxicity external quality control programme (CT-EQC). STUDY DESIGN: The results of the participating laboratories in Spanish CT-EQC programme for human reproduction laboratories during the period 2013-2016 were analyzed. Per year, laboratories receiving three pipette tips and three aliquots of culture medium. All laboratories used the human sperm survival test to perform the bioassay. On average 48 laboratories took part in the programme each year. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and overall accuracy were calculated, with the corresponding 95% confidence intervals. RESULTS: Overall, for both products, sensitivity was higher than specificity, and NPV was higher than PPV. For laboratories participating for the first time in the CT-EQC, lower results were obtained in sensitivity and specificity in culture media than in pipette tips. However, in subsequent years, these differences disappeared. The PPV obtained for pipette tips was higher than that obtained for culture media (0.82 (0.77-0.87) vs 0.71 (0.66-0.76)). No relationship was recorded between the laboratories' accuracy in culture media and pipette tips (r = 0.026). CONCLUSIONS: From a logistical standpoint, pipette tips are more appropriate than culture medium for use in a CT-EQC programme.

Effect of monotherapy with darunavir/cobicistat on viral load and semen quality of HIV-1 patients.

Many patients previously using darunavir/ritonavir (DRV/r) (800/100mg) have switched to darunavir/cobicistat (DRV/C) (800/150 mg) either as part of triple therapy (ART) or as monotherapy with DRV (mDRV). The latter approach continues to be used in some countries for patients receiving long-term treatment. However, to date, the behaviour of DRV/C in the seminal compartment has not been analysed. This study explores how the combination behaves in monotherapy, with respect to the control of viral load and seminal quality. To this end, we studied 20 patients who were treated with mDRV/C after previous treatment with mDRV/r for at least 24 weeks. A viral load control in seminal plasma similar to that published in the literature was observed after 24 weeks of treatment with mDRV/C (viral load positivity in 20% of patients). Similarly, semen quality was confirmed (70% normozoospermic) in patients treated with this formulation, as has previously been reported for ART and mDRV/r. The DRV levels measured in seminal plasma were above EC50, regardless of whether the seminal viral load was positive or negative. We conclude that this mDRV/C co-formulation behaves like mDRV/r in seminal plasma in terms of viral load control and semen quality.

Economic evaluation of elective single-embryo transfer with subsequent single frozen embryo transfer in an in vitro fertilization/intracytoplasmic sperm injection program.

OBJECTIVE: To analyze the cost-effectiveness of IVF-ICSI cycles with elective single-embryo transfer (eSET), plus elective single frozen embryo transfer (eSFET) if pregnancy is not achieved, compared with double-embryo transfer (DET). DESIGN: Cost-effectiveness analysis. SETTING: Public hospital. PATIENT(S): A population of 121 women (<38 years old), undergoing their first or second IVF cycles. INTERVENTION(S): We conducted a cost-effectiveness analysis using the results of a prospective clinical trial. The women in group 1 received eSET plus eSFET, and those in group 2 received DET. A probabilistic sensitivity analysis was performed. MAIN OUTCOME MEASURE(S): Live birth delivery rate. RESULT(S): The cumulative live birth delivery rate was 38.60% in the eSET+eSFET group versus 42.19% in the DET group. The mean costs per patient were €5,614.11 in the eSET+eSFET group and €5,562.29 in the DET group. These differences were not statistically significant. The rate of multiple gestation was significantly lower in the eSET group than in the DET group (0 vs. 25.9%). CONCLUSION(S): This study does not show that eSET is superior to DET in terms of effectiveness or of costs. The lack of superiority of the results for the eSET+eSFET and the DET groups corroborates that the choice of strategy to be adopted should be determined by the context of the health care system and the individual prognosis.

Assisted reproductive technology and obstetric outcome in couples when the male partner has a chronic viral disease.

BACKGROUND: Assisted reproductive technology (ART) with washed semen can achieve pregnancy with minimal risk of horizontal and vertical transmission of chronic viral diseases (CVD) such as human immunodeficiency virus (HIV), hepati- tis C virus (HCV) and hepatitis B virus (HBV) among serodiscordant couples. How- ever, few studies have been made of the use made by these couples of ARTs or of the obstetric results achieved. MATERIALS AND METHODS: In this retrospective study, 93 men who were seropositive for HIV, HCV or HBV and who underwent assisted reproduction treatment at our centre (Hospital Universitario Virgen de las Nieves, Granada, Spain) were included. Washed semen was tested to detect viral particles. Non-infected women were tested before and after each treatment, as were the neonates at birth and after three months. RESULTS: A total of 62 sperm samples were washed, and none were positive for the detec- tion of viral molecules. Semen samples from 34 HBV positive males were not washed since the female partner had immunity to hepatitis B. In total, 38 clinical pregnancies were achieved (22% per cycle and 40.9% per couple) out of 173 cycles initiated, and 28 births were achieved (16.2% per cycle and 30.1% per couple), producing 34 live births. The rate of multiple pregnancies was 21.4%. Obstetric and neonatal results were similar in the groups of couples studied. At follow-up, no seroconversion was detected in the women or neonates. CONCLUSION: Sperm washing and intracytoplasmic sperm injection are shown to be a safe and effective option for reducing the risk of transmission or super infection in serodiscordant or concordant couples who wish to have a child. Pregnancies ob- tained by ART in couples when the male is CVD infected achieve good obstetric and neonatal results.

Reducing inter-observer variability in embryo evaluation by means of training courses.

PURPOSE: To study the utility of a training session offered to junior embryologists, comparing the results obtained with those reported by a group of senior embryologists. METHODS: The 62 junior embryologists participanting were asked to decide on the quality of the embryos and theg clinical decision to be taken. RESULTS: The junior embryologists' success rate following the training course was significantly higher than before for embryo classification (48.4% ± 20.4 vs. 59.7% ±16.7) (p < 0.05) and for clinical decision (54.7% ± 19.6 vs. 68.7% ± 17.6) (p < 0.005). Comparison of the degree of agreement between the categories assigned by the junior embryologists and those assigned by consensus among the group of senior embryologists revealed kappa values of k = 0.32 before the course and of k = 0.54 after it. The comparison between pre- and post-training junior and senior embryologists also reflected an improvement in the kappa index for clinical decision, from k = 0.54 to k = 0.68. CONCLUSIONS: Training courses are shown to be an effective tool for increasing the degree of agreement between junior and senior embryologists.

Aplicación de las especificaciones de calidad analíticas al cálculo de los límites admisibles de error en Programas de Evaluación Externa de la Calidad para concentración espermática / Application of the analytical quality specifications to the calculation of the allowable limits of error in Programs of External Quality Assessment for sperm concentration

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PGS-FISH in reproductive medicine and perspective directions for improvement: a systematic review.

INTRODUCTION: Embryo selection can be carried out via morphological criteria or by using genetic studies based on Preimplantation Genetic Screening. In the present study, we evaluate the clinical validity of Preimplantation Genetic Screening with fluorescence in situ hybridization (PGS-FISH) compared with morphological embryo criteria. MATERIAL AND METHODS: A systematic review was made of the bibliography, with the following goals: firstly, to determine the prevalence of embryo chromosome alteration in clinical situations in which the PGS-FISH technique has been used; secondly, to calculate the statistics of diagnostic efficiency (negative Likelihood Ratio), using 2 × 2 tables, derived from PGS-FISH. The results obtained were compared with those obtained from embryo morphology. We calculated the probability of transferring at least one chromosome-normal embryo when it was selected using either morphological criteria or PGS-FISH, and considered what diagnostic performance should be expected of an embryo selection test with respect to achieving greater clinical validity than that obtained from embryo morphology. RESULTS: After an embryo morphology selection that produced a negative result (normal morphology), the likelihood of embryo aneuploidies was found to range from a pre-test value of 65% (prevalence of embryo chromosome alteration registered in all the study groups) to a post-test value of 55% (Confidence interval: 50-61), while after PGS-FISH with a negative result (euploid), the post-test probability was 42% (Confidence interval: 35-49) (p < 0.05). The probability of transferring at least one euploid embryo was the same whether 3 embryos were selected according to morphological criteria or whether 2, selected by PGS-FISH, were transferred. Any embryo selection test, if it is to provide greater clinical validity than embryo morphology, must present a LR-value of 0.40 (Confidence interval: 0.32-0.51) in single embryo transfer, and 0.06 (CI: 0.05-0.07) in double embryo transfer. DISCUSSION: With currently available technology, and taking into account the number of embryos to be transferred, the clinical validity of PGS-FISH, although superior to that of morphological criteria, does not appear to be clinically relevant.

Papel del profesional del laboratorio clínico en la desmedicalización: el ejemplo del análisis de semen y los deseos reproductivos / No disponible

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External quality control for embryology laboratories.

Participation in external quality control (EQC) programmes is recommended by various scientific societies. Results from an EQC programme for embryology laboratories are presented. This 5-year programme consisted of the annual delivery of (i) materials to test toxicity and (ii) a DVD/CD-ROM with images of zygotes and embryos on days 2 and 3, on the basis of which the participants were asked to judge the embryo quality and to take a clinical decision. A high degree of agreement was considered achieved when over 75% of the laboratories produced similar classifications. With respect to the materials analysed, the specificity was 68% and the sensitivity was 83%. Concerning embryo classification, the proportion of embryos on which a high degree of agreement was achieved increased during this period from 35% to 55%. No improvement was observed in the degree of agreement on the clinical decision to be taken. Day-3 embryos produced a higher degree of agreement (58%) than did day-2 embryos (32%) (P<0.05). Participation in EQC increased the degree of inter-laboratory agreement on embryo classification, but not the corresponding agreement on clinical decision taking. It is necessary to introduce measures aimed at standardizing decision taking procedures in embryology laboratories.

Is natural conception a valid option for HIV-serodiscordant couples?

The remarkable reduction in HIV-related morbidity and mortality as a consequence of the widespread use of highly active antiretroviral therapy (HAART) has led to a growing number of HIV-infected persons and their partners requesting counselling regarding the chances of reproduction. A thoughtful medical evaluation of the couple, which should entail HIV status, screening for genital infections and fertile potential, is needed before considering any reproductive attempt. Given that both sexual and perinatal transmission of HIV is directly correlated with the level of viral replication, being almost negligible in patients with undetectable viremia, HAART should be given to the infected partner to minimize the risk of transmission. Assisted reproduction after 'sperm washing' may further reduce the chances of infection, although this is not within reach or desire for a significant number of HIV-serodiscordant couples. From our perspective, natural conception could now be considered a possible alternative for HIV-serodiscordant couples, as long as complete suppression of viremia with HAART is achieved in the infected partner. The objective of this paper is to propose a protocol that may minimize risks in HIV-discordant couples that have opted for natural conception.

Procedure control and acceptance sampling plans for donor sperm banks: a theoretical study.

The publication of European Directive 2004/23/EC in the European Parliament and in the European Council on 31 March 2004 concerning the setting of standards of quality and safety for the donation, procurement, testing, processing, preservation, storage and distribution of human cells and tissues made it obligatory for sperm banks to set up quality control systems to ensure, among other goals, the satisfactory control of all procedures carried out. The objective of the present study is to set out guidelines that will make it possible to ensure the quality of the donors and frozen specimens accepted and the homogeneity of the samples supplied by a sperm bank. For this purpose, we shall describe clear-cut criteria for the acceptance of donors and frozen sperm, taking into account both analytic variability and the biological variations to be expected in semen parameters. Furthermore, we shall show how the evaluation of the results of a frozen semen specimen, on the basis of analysing a single straw after such freezing, does not guarantee the homogeneity of all the straws. Therefore, we must design a sampling plan to take into consideration all the straws obtained from a donor. This kind of plan will depend on different parameters, such as acceptable levels of quality and the tolerable rate of straws with defective semen, and will involve certain risks, both for the sperm bank and for the client. The establishment of these acceptance control criteria for frozen specimens and for donors could be of practical use for the control of the procedures applied in the operation of a sperm bank.

Expression of transcription factors in endometrium during natural cycles.

PURPOSE: The sex steroid control of the endometrial cycle is mediated by transcription factors, four of which are the estrogen and progesterone receptors, c-jun and c-fos, all expressed by the endometrium. The aim of this study was to analyze the distribution of the transcription factors in the different endometrial compartments during natural cycles. METHODS: We studied 53 reproductively-normal women, of whom 26 were in the proliferative phase and 27 in the secretory phase. An endometrial biopsy was performed and serum values of LH, FSH, estradiol, and progesterone were determined. We studied the expression of transcription factors using monoclonal antibodies. RESULTS: A correlation between estrogen receptor and c-jun and c-fos expression was observed in stroma and epithelia, and progesterone receptor expression correlated with c-jun expression in epithelia. C-jun and c-fos presented greater expression in the proliferative phase than in the secretory phase, in the stroma and in both epithelia. No relation was found between estradiol serum levels and any transcription factor, but progesterone serum levels correlated significantly with most such factors. CONCLUSION: The two proto-oncogenes could play a decisive role in regulating the endometrial cycle; they could mediate the effects induced by sex steroid, and could be related to other transcription factors.