Revisiting the impact of ejaculatory abstinence on semen quality and intracytoplasmic sperm injection outcomes.

BACKGROUND: Regulatory bodies recommend inconsistent ejaculatory abstinence lengths before semen analysis. The literature exploring the effect of ejaculatory abstinence length on the outcomes of intracytoplasmic sperm injection is scarce. OBJECTIVE: To study the influence of ejaculatory abstinence length on semen quality and intracytoplasmic sperm injection outcomes. MATERIALS AND METHODS: This prospective cohort study included 818 patients undergoing conventional semen analysis from October 2015 to October 2016, in a private university-affiliated IVF centre. Generalized linear models adjusted for potential confounders were used to investigate the associations between ejaculatory abstinence length and seminal parameters and intracytoplasmic sperm injection outcomes. RESULTS: Increasing ejaculatory abstinence length was positively correlated with semen volume, sperm concentration, total sperm count, total motile sperm count and sperm DNA fragmentation index. Significant inverse correlations were observed between ejaculatory abstinence length and fertilization rate, blastocyst formation rate, implantation rate and pregnancy rate. A discriminant analysis showed a mean ejaculatory abstinence length in the positive pregnancy group of 3.14 ± 1.64 days and 4.83 ± 3.66 days in the negative pregnancy group. A cut-off point was established halfway between ejaculatory abstinence length averages, at 4 days. The ejaculatory abstinence ≤4 days group showed significant lower semen volume, sperm concentration, total sperm count and total motile sperm count compared to ejaculatory abstinence > 4 days group. The ejaculatory abstinence ≤ 4 days group showed significant lower sperm DNA fragmentation index, and higher rates of fertilization, high-quality embryos on day 3, blastocyst development, implantation and pregnancy compared to ejaculatory abstinence > 4 days group. The implantation rate was significantly higher and the pregnancy rate tended to be higher with one day of ejaculatory abstinence, compared to 2-4 days of ejaculatory abstinence. CONCLUSIONS: Ejaculatory abstinence periods of >4 days have a detrimental effect on sperm DNA and intracytoplasmic sperm injection outcomes. One day of ejaculatory abstinence significantly improves implantation rate and tends to increase pregnancy rate, compared to 2, 3 and 4 days of ejaculatory abstinence.

Sperm morphological normality under high magnification is correlated to male infertility and predicts embryo development.

Human sperm morphology has been described as an essential parameter for the diagnosis of male infertility and a prognostic indicator of natural or assisted pregnancies. Nevertheless, standard morphological assessment remains a subjective analysis and its impact on intracytoplasmic sperm injection (ICSI) is also of limited value. The objective of this prospective cohort study was to investigate whether motile sperm organelle morphology examination (MSOME) can improve semen analysis by better defining male infertility and providing a better prognosis for ICSI up to a year later. Data were obtained from 483 patients undergoing conventional semen analysis from June 2015 to June 2017 in a private university-affiliated in vitro fertilization (IVF) center. The correlation of MSOME with seminal parameters was evaluated. One hundred and thirty patients underwent ICSI up to a year later, and the correlation between MSOME and ICSI outcomes was established. Except for volume, all seminal parameters were positively correlated with MSOME I+II. MSOME was also distinct between World Health Organization (WHO) classification groups, with normozoospermic and oligoasthenoteratozoospermic presenting the higher and the lower proportion of MSOME I+II, respectively. MSOME I+II was prognostic for fertilization rate, high-quality cleavage-stage embryos rate, and blastocyst rate. The normality cutoff value based on blastocyst rate was MSOME I+II≥ 5.5%. MSOME could be a useful tool for the diagnosis of infertility severity as it is correlated with sperm morphology, motility, and concentration. Men who had higher MSOME I+II had better ICSI outcomes. The future use of MSOME as a routine method for semen analysis may be a reliable form of assessing male infertility.

Total motile sperm count has a superior predictive value over the WHO 2010 cut-off values for the outcomes of intracytoplasmic sperm injection cycles.

The objective of this study was to compare (i) the intracytoplasmic sperm injection outcomes among groups with different total motile sperm count ranges, (ii) the intracytoplasmic sperm injection outcomes between groups with normal and abnormal total motile sperm count, and (iii) the predictive values of WHO 2010 cut-off values and pre-wash total motile sperm count for the intracytoplasmic sperm injection outcomes, in couples with male infertility. This study included data from 518 patients undergoing their first intracytoplasmic sperm injection cycle as a result of male infertility. Couples were divided into five groups according to their total motile sperm count: Group I, total motile sperm count <1 × 10(6) ; group II, total motile sperm count 1-5 × 10(6) ; group III, total motile sperm count 5-10 × 10(6) ; group IV, total motile sperm count 10-20 × 10(6) ; and group V, total motile sperm count >20 × 10(6) (which was considered a normal total motile sperm count value). Then, couples were grouped into an abnormal and normal total motile sperm count group. The groups were compared regarding intracytoplasmic sperm injection outcomes. The predictive values of WHO 2010 cut-off values and total motile sperm count for the intracytoplasmic sperm injection outcomes were also investigated. The fertilization rate was lower in total motile sperm count group I compared to total motile sperm count group V (72.5 ± 17.6 vs. 84.9 ± 14.4, p = 0.011). The normal total motile sperm count group had a higher fertilization rate (84.9 ± 14.4 vs. 81.1 ± 15.8, p = 0.016) and lower miscarriage rate (17.9% vs. 29.5%, p = 0.041) compared to the abnormal total motile sperm count group. The total motile sperm count was the only parameter that demonstrated a predictive value for the formation of high-quality embryos on D2 (OR: 1.18, p = 0.013), formation of high-quality embryos on D3 (OR: 1.12, p = 0.037), formation of blastocysts on D5 (OR: 1.16, p = 0.011), blastocyst expansion grade on D5 (OR: 1.27, p = 0.042), and the odds of miscarriage (OR: 0.52, p < 0.045). The total motile sperm count has a greater predictive value than the WHO 2010 cut-off values for laboratory results and pregnancy outcomes in couples undergoing intracytoplasmic sperm injection as a result of male infertility.

The negative influence of sperm cryopreservation on the quality and development of the embryo depends on the morphology of the oocyte.

The present case-control study aimed to identify the effect of sperm cryopreservation on the quality of the embryo and on the probability of blastocyst formation when oocytes free of dimorphisms are injected and when at least one dymorphism is present. The study included 22 186 zygotes, obtained from 2802 patients undergoing intracytoplasmic sperm injection cycles, in a private assisted reproduction center, using either fresh or cryopreserved sperm. The effect of sperm cryopreservation on the embryo quality on cleavage stage and blastocyst formation chance were evaluated when oocytes free of dimorphisms are injected and when at least one dymorphism is present. The quality of the embryo on cleavage stage as well as the chance for blastocyst formation was not influenced by the origin of the spermatozoa when the quality of the oocyte was not considered. When at least one oocyte defect was present, a negative influence of sperm cryopreservation on cleavage stage embryo quality and the chance for blastocyst formation was noted. In oocytes with extra-cytoplasmic dimorphisms, the injection of cryopreserved sperm did not affect the quality of the embryo during the cleavage stage, but did affect the chance for blastocyst formation. Conversely, in oocytes with intracytoplasmic defects, the quality of the embryos on cleavage stage and the chance of blastocyst formation were negatively influenced by the injection of cryopreserved sperm. The results suggest an oocyte quality-dependent negative effect of sperm cryopreservation on embryo quality and on the probability of blastocyst formation.

Intracytoplasmic morphologically selected sperm injection is beneficial in cases of advanced maternal age: a prospective randomized study.

OBJECTIVE: To evaluate advanced maternal age as a rationale for performing intracytoplasmic morphologically selected sperm injection (IMSI). STUDY DESIGN: This study included couples undergoing intracytoplasmic sperm injection (ICSI) as a result of advanced maternal age (≥37 years old). Sample size calculations were based on the assumption that a 15% difference in implantation rate would mean a clinically significant difference. To achieve this difference, 33 cycles would be needed in each treatment arm (with a significance level of 5% and power of 85%). Couples were randomly allocated to one of two sperm selection procedures (ICSI, n=33; or IMSI, n=33). Sperm selection in the ICSI group was analyzed under a magnification of 400×. Sperm selection in the IMSI group was analyzed under high magnification of 6600×. The groups were compared with regard to the outcome of the cycles. RESULTS: IMSI cycles showed significantly higher implantation (4/33, 12.1% vs. 18/47, 38.3%, p=0.026) and pregnancy (4/29, 13.8 vs. 18/30, 60.0%, p<0.001) rates. The IMSI procedure positively influenced the blastocyst formation rate (RC: 15.00, R2: 49.9%, p=0.001) and implantation rate (RC: 24.04, R2: 9.6, p=0.027), and was determinant to the increased odds of pregnancy (OR: 9.0, CI: 2.17-37.38, p=0.001). CONCLUSION: It seems that the injection of a morphologically normal spermatozoon overcomes the low oocyte quality in older women, resulting in improved embryo quality and in a 9-fold increase in the clinical pregnancy rate in couples with advanced maternal age.

The male factor of infertility should not be an issue for the selection of patients for extended embryo culture programmes.

This study evaluated the influence of sperm origin and basic sperm parameters on blastocyst implantation competence. The study included 2912 embryos obtained from 370 patients undergoing intracytoplasmic sperm injection cycles, with embryo transfer on day 5 of development. The embryos were divided into experimental groups according to their origin: (i) embryos originated from ejaculated-derived spermatozoa (Ejaculated group, n = 2093), from epididymal-derived spermatozoa (Epididymal group, n = 463) and from testicular-derived spermatozoa (Testicular group, n = 356). The groups were compared in relation to their blastocyst implantation competence. In addition, the influence of sperm parameters on blastocyst implantation was investigated. The sperm origin was determinant to the success of implantation. When blastocysts originating from testicle-derived spermatozoa were transferred, 66.4% implanted, while only 35.8 and 48.6% of blastocysts originated from epididymis- and ejaculate-derived spermatozoa implanted respectively (p = 0.001). The sperm volume and concentration were increased in cycles in which the implantation rate was 100 compared to the 0% implantation rate cases; however, the sperm motility and morphology did not differ among the groups. These results suggest that, with the exception of sperm volume and concentration, the male factor of infertility should not be an issue for the selection of patients for extended embryo culture programmes, even when azoospermic patients are considered.

Cytokine and hormonal profile in serum samples of patients undergoing controlled ovarian stimulation: interleukin-1beta predicts ongoing pregnancy.

BACKGROUND: Changes in the endometrium are not regulated exclusively by ovarian hormones; the immune system has also been implicated in normal endometrial function, similar to processes taking place during inflammatory and reparative path. Many cytokines are crucially important for reproductive processes, and the role of cytokines in the female reproductive system function has been broadly investigated during controlled ovarian stimulation (COS) for IVF attempts. The aim of this study was to evaluate the levels of serum cytokines and hormones, and the clinical outcomes of women who underwent COS and ICSI procedures. METHODS: The study prospectively included 96 patients (aged 22-43 years, unexplained or male infertility, n = 61; female infertility factors, n = 35) who underwent ICSI cycles. Serum levels of interleukin (IL-8, IL-6, IL-1beta, IL-10, IL-12), tumour necrosis factor and leukaemia-inhibitory factor (LIF) and the hormones FSH, estradiol, progesterone, anti-Mullerian hormone and Inhibin-B were measured on the day of oocyte retrieval. RESULTS: The ongoing pregnancy rate was 25.3%. The presence of serum IL-1beta positively affected the implantation rate (P = 0.004) and increased the chance of becoming pregnant by 15 fold. Furthermore, the percentage of patients with detectable serum IL-1beta levels who conceived (62.5%) was higher than those who failed to conceive (37.5%; P = 0.019). The LIF was undetectable in all serum samples, and no other factors influenced the clinical outcomes of patients undergoing ICSI cycles. CONCLUSIONS: Our findings revealed that detectable serum levels of IL-1beta on the day of oocyte retrieval in patients undergoing COS and ICSI are predictive of successful implantation and ongoing pregnancy.

Assisted reproduction professionals' awareness and attitudes towards their own IVF cycles.

OBJECTIVE: Professionals involved in assisted reproductive technologies (ART) have in-depth awareness and knowledge of the risks of multiple pregnancies at the conclusion of in vitro fertilization (IVF) treatment. The aim of the study was to investigate ART professionals' attitudes towards the awareness of the risk of infertility, as well as the decision-making process in IVF issues. METHODS: Seventy ART professionals answered a questionnaire covering demographic data, infertility awareness and attitudes towards IVF. RESULTS: Approximately half (50.8%) of the participants thought that they were not at risk of infertility. However, if they received a diagnosis of infertility, none would accept childlessness and almost all would undergo IVF. In an IVF cycle, the number of high-quality embryos transferred would be around three, but if treatment was extended to a third cycle, a higher percentage of participants would elect to transfer four or more embryos. All participants would prefer to undergo IVF and accept the risk of multiple pregnancy than remaining childless. It was found that less than a third of ART professionals considered triplets to be an unacceptable complication of IVF. CONCLUSIONS: Diagnosis of infertility affects all participants psychosocially, supporting the idea that the emotional aspects of wanting a biological child, and decision making about whether to undertake ART, outweigh the medical position regarding the risks and benefits of IVF.

Post-Transcription Regulation of mRNA and Hormone Synthesis and Release at the Anterior Pituitary Gland: Information derived from the recovery of pituitary gonadotrope cell function after with a GnRH agonist

This review presents a summary of post-transcription regulation of mRNAs with a focus on the anterior pituitary gland. The control of gene transcription and production of mRNAs is the predominant form of regulation of hormone synthesis. However, post-transcription regulation of mRNAs provides another level of control of hormone synthesis. Examples of how hormone synthesis can be controlled at the level of mRNA include mRNA nuclear export and subcellular localization, mRNA stability and turnover, and regulation of mRNA translation. The gonadotrope cells of the anterior pituitary have multiple internal effector systems and provide an ideal model cell to study post-transcription regulation of mRNAs. Gonadotrope cells are stimulated to release LH and FSH by hypothalamic GnRH that binds to GnRH receptors. GnRH receptors are coupled to G-proteins and second messenger signaling pathways that involve cAMP and IP3. These signaling pathways are associated with the release of LH and FSH and transcription of mRNAs for LH and FSH. The stability of these mRNAs can be influenced by androgens, estrogens and progestagens. Therapy with a GnRH agonist leads to desensitization of gonadotrope cells to GnRH and a depletion of cellular stores of LH and FSH mRNAs, and content of LH and FSH. After discontinuation of therapy with GnRH agonist, levels of LH and FSH mRNAs return to normal some time before LH and FSH content and secretion are restored. This is indicative of post-transcription regulation of LH and FSH mRNAs. Future studies on post-transcription regulation of mRHAs will provide new molecular insights into how gonadotrope cells balance and integrate stimulation by GnRH with feedback modulation by the gonads.