Homocysteine serum levels correlate with the number of failed IVF cycles even when within normal range.

INTRODUCTION: Repeated implantation failure is a common challenge in daily practice. Homocysteine and vitamin B12 have been associated with reproductive processes among patients undergoing in vitro fertilization; however, their involvement in repeated implantation failure has not been assessed. We explored possible associations of serum homocysteine and vitamin B12 with repeated implantation failure. MATERIAL AND METHODS: A retrospective analysis of 127 women who underwent ≥ 3 unsuccessful embryo transfers during 2005-2016, at the Fertility and In Vitro Fertilization Unit at Carmel Medical Center. After at least 3 IVF failures serum levels of homocysteine and vitamin B12 were measured. RESULTS: The mean patient age was 33.5 ± 5.2 years. The mean number of embryo transfers was 4.6 ± 1.5. The mean total cumulative number of embryos transferred was 10.4 ± 5.2. Mean serum levels of homocysteine were 8.6 ± 3.7 µM/L, and of vitamin B12 were 302.5 ± 155.3 pg/ml. Homocysteine levels were within the normal range (< 14 µM/L) in 95.8% of the patients. Yet, the levels of homocysteine correlated with both the number of failed embryo transfers (r = 0.34, p = 0.004) and the total cumulative number of transferred embryos (r = 0.36, p = 0.002). CONCLUSIONS: Our findings suggest an association between serum homocysteine levels and the occurrence of repeated implantation failure, even when homocystein levels were within the normal range. It should be studied whether nutritional supplementation to modulate serum homocysteine levels may improve treatment outcome.

Should Intra Cytoplasmic Sperm Injection (ICSI) be the primary insemination method in women undergoing IVF cycles with donor sperm?

PURPOSE: To compare efficacy of Intra Cytoplasmic Sperm Injection (ICSI) with conventional in vitro fertilization (IVF) on treatment outcome in women undergoing in vitro fertilization with donor sperm. METHODS: We examined retrospectively the outcome data from 203 patients undergoing fresh cycles of conventional IVF (cIVF) or ICSI and an additional 77 frozen-thawed embryo transfer (FET) cycles during 2003-2014, all using donor sperm. Fertilization, cleavage, pregnancy and live birth rates and number of high-quality embryos were compared between cIVF and ICSI. RESULTS: Altogether 185 women underwent 479 transfer cycles of fresh embryos (237 cIVF vs. 224 ICSI and 18 "rescue ICSI" cycles). In addition, 77 FET cycles were compared (24 cIVF vs. 53 ICSI cycles). No differences were found between cIVF and ICSI in fertilization, cleavage, pregnancy and live birth rates (92.6% vs 92.2%, 73.4% vs 72.4%, 25.3% vs 27.2% and 13.1% vs 14.7%, respectively). Pregnancy and life birth rates remained similar even when FET cycles were included (25.8% vs 26.2% and 13.1% vs 13.7%, respectively). The use of ICSI was associated with lower rates of high-quality embryos (52.7% vs. 63.3%, P < 0.0001). A multivariate logistic regression analysis found that patients' age, number of transferred embryos and smoking were independently associated with the chance to conceive. Patient age correlated inversely with fertilization rate (r = - 0.13, P < 0.006).Non-smokers were more likely to become pregnant (OR = 2.23, P < 0.012). CONCLUSIONS: Our results show that ICSI does not bypass the age-related decrease in oocyte quality in patients using donor sperm for IVF. Use of ICSI was associated with lower rates of high-quality embryos. The findings imply that ICSI should not be the primary method of insemination in patients undergoing IVF with donor sperm.

The impact of intentional endometrial injury on reproductive outcomes: a systematic review and meta-analysis.

BACKGROUND: Endometrial injury is an intentional damage made to the endometrium, usually produced by a Pipelle catheter. Over the last two decades, endometrial injury has been studied to improve implantation rates and decrease the incidence of implantation failure in invitro fertilization (IVF) cycles. Recently, additional studies of endometrial injury, performed not only in patients with implantation failure but also in intrauterine insemination cycles, have been conducted, and the endometrial injury made by hysteroscopy has been researched. The evidence describing the impact of endometrial injury is controversial; therefore, we conducted a systematic review and meta-analysis to examine the issue. OBJECTIVE AND RATIONALE: Our objective is to review the research that has been done until now and perform a meta-analysis regarding endometrial injury and its influence on implantation success and pregnancy rates in patients with at least one failed IVF cycle. In particular, we aim to study the efficacy of the procedure and look for confounding factors, such as maternal age, in assessing the efficacy of endometrial injury. SEARCH METHODS: The systematic review of the literature was conducted according to the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) statement. Study protocol can be assessed at PROSPERO International prospective register of systematic reviews (registration number CRD42018092773). Searches were conducted by an experienced research librarian in the following databases: MEDLINE(R) using the OvidSP interface and PUBMED, Embase, Web of Science and Cochrane Library. This review considered for inclusion randomized-controlled trials examining the success of performing local endometrial injury on IVF outcomes in women with previous failed IVF cycles. OUTCOMES: Ten studies, comprising a total of 1260 patients, were selected. Overall, when studying the effect of endometrial injury on clinical pregnancy rates (CPRs) and live birth rates (LBRs), higher rates were shown in the endometrial injury group. However, endometrial injury did not significantly improve CPRs and LBRs, when considering sub-group analyses of studies including patients with two or more failed IVF cycles, studies examining older patients or studies which did not include hysteroscopy. There was no significant difference found regarding multiple pregnancy rates, while a handful of studies showed an improvement in miscarriage rates. WIDER IMPLICATIONS: Endometrial injury should be used restrictively and not routinely in clinics. Maternal age and number of previous failed treatment cycles may be contributing factors which can influence the results when studying the effect of endometrial local injury. It is possible that the relative contribution of endometrial receptivity to the chances of implantation decreases with any additional failed cycle. The optimal study to prove the efficacy of local endometrial injury on implantation and pregnancy rates, should be a random-controlled trial studying the effect of local endometrial injury in oocyte donation cycles, in recipients with repeated implantation failure. This kind of study will conclude whether local endometrial injury is an efficient procedure with minimum confounding factors, and may assist in defining the population, even outside of donation cycles, that will benefit from the procedure.

Oxidative markers in cryopreservation medium from frozen-thawed embryos: a possible tool for improved embryo selection in in vitro fertilization?

PURPOSE: The present study evaluated the association between oxidative parameters in embryo cryopreservation medium and laboratory and clinical outcomes. METHODS: This prospective laboratory study was conducted in an IVF unit in a university-affiliated hospital with 91 IVF patients undergoing a frozen-thawed embryo transfer cycle. Following thawing, 50 µL of embryo cryopreservation medium was retrieved from each cryotube and tested by the thermochemiluminescence (TCL) assay. TCL amplitudes after 50 (H1), 150 (H2), and 280 s (H3) were recorded in counts per second (CPS) and the TCL ratio determined for comparison with implantation and pregnancy rates. RESULTS: A total of 194 embryos were transferred in 85 frozen-thaw cycles. Twenty-one pregnancies (24.7 %) occurred. Implantation and overall and clinical pregnancy rates were higher when the median TCL H1 amplitude was <32 CPS compared to ≥32 CPS (14.6 vs. 5.3 %, 37.5 vs. 17 %, 28.1 vs. 9.4 %, respectively). No pregnancies occurred when the H1 amplitude was ≥40 CPS. Logistic regression multivariate analysis found that only the median TCL H1 amplitude was associated with the occurrence of pregnancy (OR = 2.93, 95 % CI 1.065-8.08). The TCL ratio inversely correlated with the duration of embryo cryopreservation (r = -0.37). CONCLUSIONS: The results indicate that thawed embryos may express oxidative processes in the cryopreservation medium, and higher oxidative levels are associated with lower implantation rates. These findings may aid in the improved selection of frozen-thawed embryos for IVF.

[CHALLENGING THE OPTIMAL NUMBER OF RETRIEVED OOCYTES AND ITS IMPACT ON PREGNANCY AND LIVE BIRTH RATES IN IVF/ICSI CYCLES].

INTRODUCTION: Large numbers of retrieved oocytes are associated with higher chances of having cryopreservation of embryos. However, the process entailed exposes women to increased risk for ovarian hyperstimulation syndrome. Furthermore, mild ovary stimulation protocols are more patient-friendly and with less adverse effects. Only limited reports exist on the significance of the number of retrieved oocytes achieved in a single stimulation cycle. AIM: To investigate the optimal number of retrieved oocytes to achieve pregnancy and live birth. METHODS: This retrospective analysis included 1590 IVF cycles. Oocytes maturation, fertilization, cleavage, as well as pregnancy and live birth rates were analyzed according to the number of retrieved oocytes. RESULTS: Oocyte maturation, fertilization and cleavage rates were lower in cycles with more than 10 retrieved oocytes compared with other groups. Live birth rates were highest when the number of retrieved oocytes was 11-15. CONCLUSIONS: Retrieval of more than 15 oocytes was not associated with a significant increase in chances of conception and birth. DISCUSSION: The better oocyte quality with 10 or less oocytes retrieved could be the result of a possible interference with the natural selection, or the minimized exposure of growing follicles to the potentially negative effects of ovarian stimulation. Although the average number of available embryos was higher when more than 10 oocytes were retrieved, achievement of more than 15 oocytes did not improve IVF outcome in terms of pregnancy and delivery rates. SUMMARY: Analysis of 1590 IVF cycles including the frozen-thawed transfers shows that the best outcomes were achieved with an optimal number of 11-15 oocytes.

Habits of cell phone usage and sperm quality - does it warrant attention?

Male infertility constitutes 30-40% of all infertility cases. Some studies have shown a continuous decline in semen quality since the beginning of the 20th century. One postulated contributing factor is radio frequency electromagnetic radiation emitted from cell phones. This study investigates an association between characteristics of cell phone usage and semen quality. Questionnaires accessing demographic data and characteristics of cell phone usage were completed by 106 men referred for semen analysis. Results were analysed according to WHO 2010 criteria. Talking for ≥1 h/day and during device charging were associated with higher rates of abnormal semen concentration (60.9% versus 35.7%, P < 0.04 and 66.7% versus 35.6%, P < 0.02, respectively). Among men who reported holding their phones ≤50 cm from the groin, a non-significantly higher rate of abnormal sperm concentration was found (47.1% versus 11.1%). Multivariate analysis revealed that talking while charging the device and smoking were risk factors for abnormal sperm concentration (OR = 4.13 [95% CI 1.28-13.3], P < 0.018 and OR = 3.04 [95% CI 1.14-8.13], P < 0.027, respectively). Our findings suggest that certain aspects of cell phone usage may bear adverse effects on sperm concentration. Investigation using large-scale studies is thus needed.

Advanced paternal age and reproductive outcome.

Women have been increasingly delaying the start of motherhood in recent decades. The same trend is seen also for men. The influence of maternal age on fertility, chromosomal anomalies, pregnancy complications, and impaired perinatal and post-natal outcome of offspring, has been thoroughly investigated, and these aspects are clinically applied during fertility and pregestational counseling. Male aging and reproductive outcome has gained relatively less attention. The purpose of this review is to evaluate updated and relevant literature on the effect of paternal age on reproductive outcome.

[Oxidation and female reproduction: the good, the bad and what's between].

Reactive oxygen and nitrogen species constitute an inseparable part of aerobic life on earth. They have been known to science for about 90 years, but only during the last 50 years research in this field has expanded. Initially, scientists focused on the free radicals-induced damage to biological systems. Since the eighties, a new concept has emerged, namely, that alongside the oxidative stress-induced deleterious effect and its association with a variety of diseases, a certain threshold level of oxidation is essential to intracellular signaL transduction. Recently, some data has accumulated regarding the involvement of oxidative processes in various aspects of female reproduction, including ovarian function, fertilization, early development of the embryo and implantation. Nevertheless, there is still a long way before comprehensive and thorough understanding of their role, both at the molecular level and the expression in the clinical setup of fertility patients can be achieved. In this article, we shall address some molecular biochemical processes involved in the activity of free radicals, and review the present knowledge regarding their role in female fertility, including ovarian physiology, follicular and oocyte maturation, development of the early embryo and implantation, as well as their association with reproductive pathologies such as endometriosis, polycystic ovary disease and recurrent pregnancy loss.

Oxidative parameters of embryo culture media may predict treatment outcome in in vitro fertilization: a novel applicable tool for improving embryo selection.

OBJECTIVE: To examine whether the oxidative status of an individual embryo before transfer may predict chances of implantation. DESIGN: A prospective laboratory study. SETTING: An IVF unit in a university-affiliated hospital. PATIENT(S): One hundred thirty-three women undergoing IVF-ET treatment cycles. INTERVENTION(S): Before ET, 10 µL of embryo culture medium was retrieved individually from each embryo and the oxidative status assessed by the thermochemiluminescence (TCL) analyzer. MAIN OUTCOME MEASURE(S): The occurrence of pregnancy. Two parameters were recorded: the TCL amplitude after 50 seconds (H1) and the TCL ratio. These were compared with demographic, clinical, and laboratory parameters and treatment outcome. All data underwent statistical analysis. RESULT(S): Altogether 284 embryos were transferred in 133 ET cycles. Forty-one pregnancies occurred (31%). For embryos transferred after 72 hours (77 transfers), the highest H1 levels in each group of transferred embryos correlated with the occurrence of pregnancy. The combination of maximal intracohort H1 level <210 counts per second with a TCL ratio of ≤ 80% had a positive predictive value of 70.6% for the occurrence of pregnancy. CONCLUSION(S): The oxidative status of the early embryo in IVF is associated with the chances of implantation. Assessment of the oxidative status of embryos in culture media before transfer may serve as an applicable tool for improving embryo selection in light of the legal limitations of the number of transferred embryos allowed.

Early embryo-endometrial signaling modulates the regulation of matrix metalloproteinase-3.

OBJECTIVE: To examine matrix metalloproteinase-3 (MMP-3) expression in human stromal cell culture after P stimulation and the effect of conditioned medium from human embryo-epithelial cells coculture on its expression and activity. DESIGN: Metabolic and endocrine studies on human tissue. SETTING: In vitro fertilization (i.v.f.) unit and endocrine research unit. PATIENT(S): Infertile patients undergoing endometrial tissue sampling for dating at the luteal phase before i.v.f. INTERVENTION(S): Endometrial sampling and collection of human embryos culture media. MAIN OUTCOME MEASURE(S): Expression and activity of secreted MMP-3 by P-induced stromal cells, and in stromal cells exposed to conditioned medium from embryo-epithelial cell coculture. RESULT(S): Expression and activity of MMP-3 in human stromal cells decrease after P induction. Following incubation of these stromal-derived decidual cells with conditioned medium from embryo-epithelial cell coculture, MMP-3 expression and activity increased in a statistically significant manner. CONCLUSION(S): Progesterone inhibition of MMP-3 expression and its support of endometrial integrity were prevented by local expression of MMP-3 in response to embryonic signaling.

Oxidative stress indices in follicular fluid as measured by the thermochemiluminescence assay correlate with outcome parameters in in vitro fertilization.

OBJECTIVE: To evaluate oxidative stress indices in follicular fluid (FF) by a novel thermochemiluminescence (TCL) assay and investigate the correlation between TCL and i.v.f. cycle parameters. DESIGN: Prospective, cross-sectional study. SETTING: I.v.f. Unit of an Obstetrics and Gynecology Department in a university-affiliated hospital. PATIENT(S): One hundred eighty-nine women undergoing consecutive i.v.f. treatment cycles during 2001. INTERVENTION(S): After oocyte retrieval, pooled FF was centrifuged and the supernatant was tested in the TCL assay. MAIN OUTCOME MEASURE(S): Maximal serum E2 levels, number of gonadotropin ampoules, retrieved oocytes, mature oocytes, fertilization and cleavage rates, number of available embryos and cryopreserved embryos, and clinical pregnancy rates and correlation with TCL indices. RESULT(S): The TCL curve slope of FF positively correlated with maximal serum E2 levels, number of mature oocytes, and number of cleaved embryos and was inversely correlated with the women's ages and the number of gonadotropin ampoules. Follicular fluid TCL amplitude at 50 seconds ranged from 294 to 711 cps, but all pregnancies (n = 50; 28.1%) occurred within the range of 347-569 cps. With 385 and 569 cps as cutoff levels for the occurrence of pregnancy, the negative predictive value beyond this range was 96% and the positive predictive value within this range was 32%. CONCLUSION(S): The TCL results may reflect the age-related increase in free radical activity and is associated with parameters of ovarian responsiveness and IVF outcome. A certain threshold of oxidative stress may be required for the occurrence of conception in i.v.f. TCL is a potential tool to evaluate, treat, and monitor antioxidant therapy in i.v.f. treatments.

Oxidative stress indices in seminal plasma, as measured by the thermochemiluminescence assay, correlate with sperm parameters.

OBJECTIVE: To examine the effect of oxidation of proteins and lipids, as measured by a novel thermochemiluminescence (TCL) analyzer, and to evaluate the correlation between TCL indices in seminal plasma and sperm parameters. DESIGN: Experimental and prospective clinical studies. SETTING: An infertility unit. PATIENT(S): One hundred forty-eight men undergoing semen analysis. INTERVENTION(S): Bovine serum albumin (BSA) and linolenic acid were oxidized and tested by TCL, protein carbonyls, and conjugated dienes assays. All participants underwent semen analysis. Seminal plasma was tested by TCL and conjugated dienes. MAIN OUTCOME MEASURE(S): Thermochemiluminescence indices before and after oxidation of BSA and linolenic acid, compared with protein carbonyl and conjugated dienes indices. Correlation between semen parameters and TCL and conjugated dienes indices in seminal plasma. RESULT(S): Oxidation of BSA and linolenic acid was marked by characteristic changes in their TCL curve pattern and an increase in the levels of protein carbonyls and conjugated dienes. Among 125 sperm-containing semen samples, the TCL curve exhibited two patterns: a positive relative ratio curve (group A, 87 patients) and a negative relative ratio curve (group B, 38 patients). Sperm concentration was lower and total motile sperm and rapid motile sperm were fewer in group B. A significant correlation was found between TCL indices, conjugated dienes, and sperm quality in group B. CONCLUSION(S): Oxidation affects TCL curve pattern of proteins and lipids in a characteristic manner. Thermochemiluminescence indices in seminal plasma closely correlate with sperm characteristics among patients with sperm disturbances, and it might serve as a tool in the evaluation, treatment, and monitoring of subfertile men.

Immunosuppressive properties of follicular fluid and media conditioned by zygotes correlate with subsequent conception in in vitro fertilization.

PROBLEM: Our objective in this study was to correlate immunosuppressive properties of follicular fluid (FF) and media conditioned by zygotes and early embryos with the occurrence of conception in an in vitro fertilization (IVF) program. METHOD OF STUDY: Fifty-seven IVF patients were studied. Donor lymphocytes were incubated with mitogens and FF from mature oocytes or conditioned media from zygotes and early embryos. Proliferation was assessed by radioactive thymidine incorporation. Proliferation Index (PI) was the ratio between radioactive labeling of lymphocytes incubated in the presence and absence of a mitogen. RESULTS: The FFs and media conditioned by zygotes from conception cycles had higher immunosuppressive activity than those from non-conception cycles. CONCLUSIONS: Immunosuppressive activity present in FF and media conditioned by zygotes may be a major determinant of conception in IVF, and may serve as a marker for embryo quality.