Efficacy and safety of follitropin alpha biosimilars compared to their reference product: a Meta-analysis.

AIM: Recently published multicentre, randomized phase III studies suggested the therapeutic equivalence of biosimilar follitropin alpha medicaments compared to the reference product. The aim of this meta-analysis is to pool the results of the three phase III trials in order to provide an overall analysis about the clinical bioequivalence between biosimilars and the originator. METHODS: The studies included in the analysis were pooled together in order to estimate the log odds ratio (OR) for binary outcomes and the weighted mean difference (WMD) for continuous outcomes along with the corresponding 95% confidence intervals (CI) by using a random effects model. The heterogeneity between the studies was evaluated with the Higgins and Chi-square tests. RESULTS: No differences were found in term of number of oocytes retrieved at ovum pick-up, the primary endpoint recommended by the European Medicines Agency. No statistical differences were also found for biochemical pregnancy rate, take home baby rate, total follitropin alpha dose, duration of stimulation, and OHSS risk. A significantly higher clinical pregnancy rate (p = .03) was observed in the originator group in comparison to the biosimilar follitropin alpha. CONCLUSION: Biosimilar follitropin alpha medicaments resulted comparable in comparison to the reference product with respect to the number of oocytes retrieved, that is the primary endpoint recommended by the European Medicines Agency .Further study is needed to evaluate the therapeutic bioequivalence between follitropin alpha biosimilar and the reference medication with respect to secondary endpoints.

Impact of Cigarette Smoking on the Expression of Oxidative Stress-Related Genes in Cumulus Cells Retrieved from Healthy Women Undergoing IVF.

The female reproductive system represents a sensitive target of the harmful effects of cigarette smoke, with folliculogenesis as one of the ovarian processes most affected by this exposure. The aim of this study was to analyze the impact of tobacco smoking on expression of oxidative stress-related genes in cumulus cells (CCs) from smoking and non-smoking women undergoing IVF techniques. Real time PCR technology was used to analyze the gene expression profile of 88 oxidative stress genes enclosed in a 96-well plate array. Statistical significance was assessed by one-way ANOVA. The biological functions and networks/pathways of modulated genes were evidenced by ingenuity pathway analysis software. Promoter methylation analysis was performed by pyrosequencing. Our results showed a down-regulation of 24 genes and an up-regulation of 2 genes (IL6 and SOD2, respectively) involved in defense against oxidative damage, cell cycle regulation, as well as inflammation in CCs from smoking women. IL-6 lower promoter methylation was found in CCs of the smokers group. In conclusion, the disclosed overall downregulation suggests an oxidant-antioxidant imbalance in CCs triggered by cigarette smoking exposure. This evidence adds a piece to the puzzle of the molecular basis of female reproduction and could help underlay the importance of antioxidant treatments for smoking women undergoing IVF protocols.

Successful pregnancy following the transfer of re-vitrified twice-warmed embryos due to the forced cancellation of the primary FET: A case report.

PURPOSE: Embryo cryopreservation represents a central procedure in in-vitro fertilization (IVF) programs. This report documents a Case of a successful pregnancy following the replacement of embryos that had to be re-vitrified due to the forced cancellation of the frozen embryo-transfer (FET). PRINCIPLE RESULTS: The 37- year-old patient was referred to our Assisted Reproductive Technology (ART) unit for idiopathic infertility and recurrent implantation failures. The collection cycle resulted in 8 grade-A cleavage embryos (8-10 blastomeres), that were all vitrified to prevent ovarian hyperstimulation syndrome (OHSS). The first frozen embryo transfer (FET) ended in a biochemical pregnancy and the second in an ectopic pregnancy. In the third attempt, three embryos were warmed but the provider could not complete the transfer due to cervical stenosis. The two surviving embryos were therefore re-vitrified. The final FET attempt, 4 months later, was successful and ended with the live birth of a healthy female baby. CONCLUSIONS: The transfer of re-vitrified twice-warmed embryos may represent a possible option when embryo transfer cannot be performed.

Whole-body exposure to cigarette smoke alters oocyte miRNAs expression in C57BL/6 mice.

Cigarette smoke is toxic for the female reproductive system with particular reference to the ovary. The purpose of the study was to investigate if the microRNAs (miRNAs) pattern could be altered by cigarette smoke exposure in mouse oocytes. For this purpose, C57BL/6 mice were whole-body exposed to three cigarettes daily, 7 days/week, for 2 or 4 months by a specific rodent ventilator. Mice were then superovulated and oocytes collected. MII oocytes pools obtained by single animals were deprived of cumulus cells and used to extract total RNA including miRNAs. TaqMan™ Rodent MicroRNA A Array v2.0 was used to analyze the miRNAs expression profile. The biological functions and the functional networks of the identified up- and downregulated miRNAs were analyzed by ingenuity pathway analysis software. The gene expression of deregulated-miRNAs targets was evaluated. For the first time, the global miRNAs changes in mouse oocyte in response to cigarette smoke exposure were disclosed. Our results revealed significant modulation of miRNAs mainly involved in inflammatory processes, cellular proliferation, and apoptosis. miRNAs expression was altered in a time-dependent manner. Smoke exposure induced an early downregulation of Dicer1. Transcriptional alterations of the modulated miRNAs major targets, estrogen receptor 1, peroxisome proliferator-activated receptor-alpha, and tumor protein 53, as well as that of other key regulatory genes, were evidenced. Cigarette smoke represents a stimulus able to alter miRNAs pattern in mouse oocyte. This study increases our understanding of the ovarian toxicity profile of cigarette smoke, and open new roads toward the identification of biomarkers of oocyte toxicity and dysregulation.

Cigarette smoke is associated with altered expression of antioxidant enzymes in granulosa cells from women undergoing in vitro fertilization.

This study was undertaken to evaluate whether cigarette smoke is associated with changes in the expression of antioxidant enzymes in granulosa cells of women undergoing IVF treatments. For this aim, the expression of three antioxidant enzymes (SOD1, SOD2 and catalase) in non-smokers (n = 20) and smokers (n = 20) was analyzed. There was a statistically significant overexpression of SOD2 and catalase mRNA levels in smokers in comparison with non-smokers. Cigarette smoking was associated with a lower fertilization rate, implantation rate and pregnancy rate in comparison with non-smokers. There was no effect on retrieved oocytes number, metaphase II oocytes number, quality of embryos transferred and live birth rate. These findings suggest that cigarette smoke initiates oxidative stress in granulosa cells.

Nutrition, female fertility and in vitro fertilization outcomes.

The investigation of modifiable factors that may exert influences on female reproductive health and in vitro fertilization (IVF) outcomes is increasing. Growing attention is being paid to nutrition. The aim of the present review is to recapitulate the current understanding on the effects of nutrition on female fertility and IVF outcomes. In particular, the three main classes of macromolecules have been analysed i.e. carbohydrates, proteins and fatty acids. An increasing number of studies have focused on the potential benefit of whole grain, vegetables and omega-3 polynsatured fatty acids (ω-3 PUFAs) on reproductive outcomes. Controversial results exist regarding the consumption of omega-6 (ω-6) PUFAs and dairy. Overall, nutrition appears to represent a modifiable factor that may play a significant role in the context of female reproduction and IVF outcomes, but the limited number of studies and the discrepancies between the available data call for further research in the area.

The Aftermath of Long-Term Cigarette Smoking on Telomere Length and Mitochondrial DNA Copy Number in Human Cumulus Cells Prior to In Vitro Fertilization-A Pilot Study.

Cigarette smoking among women of reproductive age is known to take a toll on systemic health and fertility potential by severely impacting ovarian tissues and cells, such as granulosa and cumulus cells (CCs). The purpose of this study was to determine the potential damage caused by tobacco smoke at a molecular level in the CCs of females who had undergone in vitro fertilization. The level of intracellular damage was determined by estimating the average telomere length (TL) and mitochondrial DNA copy number (mtDNA-CN), as well as the expression profile of telomere maintenance genes TERF1, TERF2, POT1 and microRNAs miR-155, miR-23a and miR-185. Western blotting analysis was performed to detect consequent protein levels of TERF1, TERF2 and POT1. Our results evidenced significantly lower relative TL and mtDNA-CN and a down-regulation pattern for all three described genes and corresponding proteins in the CCs of smokers compared with controls (p < 0.05). No significant differences were found in the miRNAs' modulation. Combined, our data add another piece to the puzzle of the complex regulatory molecular networks controlling the general effects of tobacco smoke in CCs. This pilot study extends the until now modest number of studies simultaneously investigating the mtDNA-CN and TL pathways in the human CCs of smoking women.

Novel Insights on the Role of Nitric Oxide in the Ovary: A Review of the Literature.

Nitric oxide (NO) is formed during the oxidation of L-arginine to L-citrulline by the action of multiple isoenzymes of NO synthase (NOS): neuronal NOS (nNOS), endotelial NOS (eNOS), and inducible NOS (iNOS). NO plays a relevant role in the vascular endothelium, in central and peripheral neurons, and in immunity and inflammatory systems. In addition, several authors showed a consistent contribution of NO to different aspects of the reproductive physiology. The aim of the present review is to analyse the published data on the role of NO within the ovary. It has been demonstrated that the multiple isoenzymes of NOS are expressed and localized in the ovary of different species. More to the point, a consistent role was ascribed to NO in the processes of steroidogenesis, folliculogenesis, and oocyte meiotic maturation in in vitro and in vivo studies using animal models. Unfortunately, there are few nitric oxide data for humans; there are preliminary data on the implication of nitric oxide for oocyte/embryo quality and in-vitro fertilization/embryo transfer (IVF/ET) parameters. NO plays a remarkable role in the ovary, but more investigation is needed, in particular in the context of human ovarian physiology.

In Vivo Cigarette Smoke Exposure to Examine the Expression of Genes Involved in the Inflammatory Response in the Mouse Uterus.

Cigarette smoke may impair uterine function, but the underlying mechanisms are poorly characterized. In this article, we describe the methodology for whole-body exposure to cigarette smoke together with assessment of the impact of this exposure on the expression of a panel of genes related to stress and toxicity pathways in mouse uteri using an in vivo model. C57BL/6 mice are whole-body-exposed to three cigarettes daily, 7 days/week, for 2 months using a specific rodent ventilator. Uteri are then collected and subjected to qRT-PCR analysis using the Stress & Toxicity PathwayFinder RT2 Profiler PCR Array (Qiagen). Cigarette smoke was found to be associated with an upregulation (≥2-fold) of C-reactive protein (Crp; 2.65-fold, p-value = 0.02), growth arrest and DNA-damage-inducible45γ (Gadd45γ; 2.11-fold, p-value = 0.04), interferon γ (Ifnγ; 2.05-fold, p-value = 0.01), and interleukin1α (Il1α; 7.74-fold, p-value = 0.003) and downregulation of matrix metallopeptidase-9 (Mmp9; -2.42-fold, p-value = 0.01). The protocol used in this study may represent a new experimental model of mouse in vivo mainstream exposure to cigarette smoke. In addition, the resulting overexpression of pro-inflammatory cytokines and genes involved in cell cycle proliferation, together with the downregulation of extracellular matrix metallopeptidases, may represent a toxicological response to cigarette smoke exposure, with potential repercussion for the processes of uterine remodeling and growth that are essential for uterine receptiveness. A recommendation to expand upon this research area is made. © 2021 Wiley Periodicals LLC.

Maternal use of fluconazole and congenital malformations in the progeny: A meta-analysis of the literature.

Fluconazole is a bis-triazole agent used in the treatment of superficial and systemic fungal infections, with vaginal candidiasis being one of the commonest indications to fluconazole treatment. There is increasing concern regarding the teratogenic potential of fluconazole. The aim of this meta-analysis is to pool the literature data in order to evaluate the possible association between fluconazole exposure during pregnancy and birth defects. A total of nine studies were included in the meta-analysis. Results were expressed as odds ratios (OR) with 95 % confidence intervals (CI) and statistical heterogeneity between the studies was evaluated with Higgins index (I2) and Q-test (Q). A p-value < 0.05 referred to the effect was considered statistically significant. The maternal exposure to fluconazole during the first trimester of pregnancy is correlated with increased prevalence of heart defects in the offspring for both low dose (OR 1.95, 95 % CI 1.18-3.21; P = 0.01) and any dose (OR 1.79, 95 % CI 1.18-2.71; P = 0.01). No association was found between gestational exposure to fluconazole and increased risk of spontaneous abortion or stillbirth. Fluconazole should be regarded as a human teratogen and should be cautiously prescribed to pregnant women and to women of childbearing potential.

Effects of Supplementation with Natural Antioxidants on Oocytes and Preimplantation Embryos.

For most infertile couples, in vitro fertilization (IVF) represents the only chance to conceive. Given the limited success of IVF procedures, novel approaches are continuously tested with the aim of improving IVF outcomes. Growing attention is devoted today to the potential benefit of natural antioxidants in the optimization of infertility treatments. This review summarizes current data in this context, focusing on both experimental studies on oocytes/embryos and clinical trials on antioxidants supplementation. Based on information gained from experimental studies, antioxidant supplementation may have beneficial effects on IVF outcomes in terms of quality and cryotolerance of in vitro produced embryos, together with positive effects on in vitro maturation oocytes and on early embryonic development. Unfortunately, from the clinical side, there is a paucity of evidence favoring the protective qualities of antioxidants. Among the antioxidants considered, coenzyme Q10 may be regarded as one of the most promising for its positive role in rescuing the oxidative stress-induced damages, but further data are needed. It is concluded that further trials are necessary to characterize the potential clinical value of antioxidants in IVF treatments.

Dynactin pathway-related gene expression is altered by aging, but not by vitrification.

The storage of surplus oocytes by cryopreservation (OC) is a widely used tool in assisted reproductive technology, but there is a great debate about the effects of cryopreservation on oocyte competence. It is known that OC may affect meiotic spindles but remains unclear if OC may increase the risk of aneuploidy. The aim of this study was to test the effects of OC and women aging on the expression of cytokinesis-related genes playing an important role in the segregation of chromosomes (DCTN1, DCTN2, DCTN3, DCTN6 and PLK1). Results highlighted that OC do not modify the expression of the selected genes, whereas women aging modulate the expression of all transcripts, confirming that aging is the crucial factor affecting meiosis and aneuploidy risk. A new role for Dynactin and PLK1 was shed in light, providing information on the ageing process in the oocyte which may be associated to reduced fertility.

Cigarette smoking impairs clinical outcomes of assisted reproductive technologies: A meta-analysis of the literature.

There is convincing evidence that cigarette smoking can impair female reproductive potential. This meta-analysis updates the knowledge regarding the effects of cigarette smoking on clinical outcomes of assisted reproductive technologies (ART). Twenty-six studies were included in this meta-analysis. Results were expressed as odds ratios (OR) with 95% confidence intervals (CI) and statistical heterogeneity between the studies was evaluated with Higgins (I2), Breslow (τ2), Birge's ratio (H2) indices and Chi-square test (χ2). A P-value < 0.05 was considered statistically significant. The analysis showed a significant decrease in live birth rate per cycle for smoking patients (OR 0.59, 95% CI 0.44-0.79; P = 0.0005), a significant lower clinical pregnancy rate per cycle for smoking women (OR 0.53, 95% CI 0.41-0.68; P < 0.0001), and a significant increase in terms of spontaneous miscarriage rate (OR 2.22, 95% CI 1.10-4.48; P = 0.025) for smokers. These findings demonstrate clear negative effects of cigarette smoking on the outcome of ART programs.

Ovotoxicity of cigarette smoke: A systematic review of the literature.

This study reviews the scientific literature on the noxious effects of cigarette smoke on the ovarian follicle, and the cumulative data on the impact of smoking on in vitro fertilization (IVF) cycle outcome. There is a close association between tobacco smoke and accelerated follicle loss, abnormal follicle growth and impairment of oocyte morphology and maturation. There is an increasing amount of evidence indicating that smoke can directly derange folliculogenesis. Increased cellular apoptosis or autophagy, DNA damage and abnormal crosstalk between oocyte and granulosa cells have been implicated in the demise of ovarian follicles. It becomes increasingly clear that maternal smoking can exert multigenerational effects on the ovarian function of the progeny. Growing evidence suggests that cigarette smoke is associated with decreased results after IVF. Further research is needed to better define the molecular mechanisms behind smoking-induced ovarian disruption.