Altered redox status may contribute to aberrant folliculogenesis and poor reproductive outcomes in women with polycystic ovary syndrome.

BACKGROUND: Women with polycystic ovary syndrome (PCOS) are often infertile and opt for artificial reproductive techniques (ART) to conceive. Disrupted pro-/antioxidant balance in oocyte microenvironment may contribute towards sub-optimal oocyte/embryo quality and poor ART outcome in them. METHODS: Activities/levels of redox markers and their transcript expression were investigated in follicular fluid and granulosa cells respectively, in women with PCOS (n = 71) and controls (n = 50) undergoing in vitro fertilization (IVF). Correlation analysis of redox markers and IVF parameters was performed. RESULTS: Activities of superoxide dismutase, glutathione reductase, glutathione peroxidase, and paraoxonase1 were significantly lower in follicular fluid of PCOS women than in controls. Levels of lipid peroxidation, oxidative protein modification, and oxidized glutathione were higher, whereas those of total antioxidant capacity, total thiols, and reduced glutathione were lower in follicular fluid of PCOS women than in controls. Further, comparison of redox markers based on insulin resistance and BMI status of study participants showed similar trends, indicating that PCOS pathophysiology is a significant contributor to oxidative stress irrespective of insulin resistance and BMI. Transcript levels of antioxidant enzymes were lower in granulosa cells from PCOS women than in controls, and they accorded with their activities in follicular fluid. Moreover, few redox markers showed significant correlations with oocyte/embryo quality and pregnancy outcome. CONCLUSION: Our data indicates disrupted redox homeostasis in follicular environment in PCOS which may negatively influence oocyte/embryo quality. Further, granulosa cells may play crucial role in maintaining follicular redox homeostasis. Glutathione system and paraoxonase1 could be explored further as surrogates for IVF prognosis/outcome.

Ovarian stimulation outcomes among transgender men compared with fertile cisgender women.

PURPOSE: To compare assisted reproductive technology (ART) outcomes among transgender men with those of fertile cisgender women. METHODS: This retrospective cohort study included 12 transgender men, six with no testosterone exposure and six after testosterone treatment, and 12 cisgender women (oocyte donors) who underwent ART in our institution between June 2017 and December 2019. Statistical analyses compared ART data and outcomes between three groups: cisgender women, transgender men without testosterone exposure, and transgender men after testosterone exposure. Comparisons were also made between transgender men with and without testosterone exposure. RESULTS: The transgender men with no testosterone exposure (23.3 ± 4 years) were significantly younger than the transgender men who had undergone testosterone treatment (30.3 ± 3.8 years; P = 0.012) and the cisgender women (29.1 ± 3.1 years; P = 0.004). The amount of FSH used for ovulation induction (1999 ± 683 mIU/mL) was significantly lower among transgender men without prior testosterone exposure compared with that among cisgender women (3150 ± 487 mIU/mL; P = 0.007). There were no differences in the peak estradiol levels, the number of oocytes retrieved, the number of MII oocytes, and the oocyte maturity rates between the three groups. Five out of six testosterone-treated transgender men underwent embryo cryopreservation, and they all achieved good-quality embryos. CONCLUSIONS: Transgender men have an excellent response to ovulation stimulation even after long-term exposure to testosterone. Oocyte/embryo cryopreservation is, therefore, a feasible and effective way for them to preserve their fertility for future biological parenting.

The role of fertility preservation in patients with endometriosis.

Patients affected with severe endometriosis are at significant risk for ovarian tissue damage, which may lead to infertility, reduced response to ovarian stimulation, and occasionally, premature ovarian failure. The risk for a compromised ovarian reserve in young patients is especially high following repeated surgical intervention and in the presence of bilateral endometriomas. In many cases, enhanced loss of ovarian reserve may also result from the damaging effect of the pathologic process on follicle reservoir even without surgical interventions. Women diagnosed with severe endometriosis and those designated for extensive ovarian surgical intervention are frequently not planning to conceive. In light of recent advances in fertility preservation techniques (FPT), such as oocytes and ovarian tissue freezing, as well as their increasing success rates, we critically evaluate the options for FPT in patients suffering from endometriosis. Personalized counseling should be offered to all patients with endometriosis taking into account age, extent of ovarian involvement, current ovarian reserve, previous and impending surgeries for endometriosis, along with current success rates and possible risks associated with FPT.

Impact of negative energy balance and postpartum diseases during the transition period on oocyte quality and embryonic development in dairy cows.

Transition period is a critical time for dairy cows because a large proportion of clinical and subclinical diseases are observed in the first month after parturition. Occurrence of negative energy balance is associated with depressed immunity and these conditions can affect oocyte quality and further embryonic development. The aim of this study was to assess the effects of negative energy balance-associated disorders on in vitro embryo production (IVP) in dairy cattle. We hypothesized that subclinical metabolic and/or inflammatory disorders have a negative effect on oocyte developmental competence and morphokinetic parameters of the resulting embryos. The study was conducted on 30 lactating Holstein-Friesian cows which were assigned into four groups: healthy (HEAL, n = 6), metabolic disease (META, n = 8), inflammatory disease (INFL, n = 8), or combined metabolic and inflammatory disease (COMB, n = 8). Ovum pick-up (OPU) was performed twice weekly on all cows over a period of four weeks (n = 8 OPU sessions/cow) starting on the fifth week postpartum, and the collected oocytes were subjected to routine IVP. Donor's health status did not affect the number of oocytes/OPU or the recovery rate (p > 0.05). The number of quality 1 oocytes collected from INFL and COMB cows was lower compared to HEAL cows (p < 0.05). Also, the percentage of quality 1 embryos was reduced in META and COMB compared to HEAL cows (p < 0.05). Cleavage, blastocyst and hatching rates were similar among groups (p > 0.05). Presence of disease did not affect the time required by zygotes to reach specific developmental stages, as recorded by means of time-lapse monitoring. Nevertheless, there was a higher probability of direct cleavage after IVF in oocytes of COMB cows compared to those of HEAL cows (p < 0.05). In conclusion, oocytes and embryos derived from dairy cows diagnosed with subclinical metabolic and/or inflammatory diseases during the transition period showed reduced quality but similar developmental potential and morphokinetics when compared to healthy cows. These results shed light on the consequences of subclinical disease on embryonic development in dairy cows which might be important for embryo transfer programs.

Enhanced Synergistic-Antioxidant Activity of Melatonin and Tretinoin by Co-encapsulation into Amphiphilic Chitosan Nanocarriers: During Mice In Vitro Matured Oocyte/Morula-Compact Stage Embryo Culture Model.

The use of exogenous antioxidants or the combination of them during in vitro oocyte/embryo culture media is reasonable. Co-delivery by nanocarrier has been designed to overcome the limitations of combining them traditionally. In this work, amphiphilic chitosan nanocarrier (ACN) was applied to co-encapsulate melatonin (Mel) and tretinoin (TTN) by the self-assembled method and evaluate their synergistic antioxidant efficacy in mice oocytes/embryos. The formation of single/dual-ACN was confirmed by Fourier-transformed infrared spectroscopy (FT-IR). The average particle diameter, size distribution, polydispersity index (PDI), and zeta potential of them were measured by dynamic light scattering (DLS), and the morphology was evaluated by TEM and SEM technologies. Also, the encapsulation efficiency (EE%) and drug loading content (DL%) of the nanocapsules were determined by UV-vis spectrophotometry. Studies of the in vitro release showed a continued drug release without any bursting effect of Mel+TTN-ACNs compared with single Mel/TTN-ACNs. Then, in both experiments, nuclear staining (Aceto-orcein and Hoechst 33342), fluorescent staining of H2DCFDA, chemiluminescence test, and qRT-PCR technique were performed as in vitro toxicity studies. The results of all these evaluations demonstrated that the dual delivery of Mel and TTN could accumulate a safety (without high-dose toxicity) synergistic anti-oxidative effect in oocyte/embryo by passive controlled, and inhibit intra/extracellular ROS levels by an enhanced intracellular penetration.

Co-encapsulation of tertinoin and resveratrol by solid lipid nanocarrier (SLN) improves mice in vitro matured oocyte/ morula-compact stage embryo development.

As a promising strategy in overcoming drug resistance, the nano drug co-delivery system (NDCDS) can transport two or more drugs into the cell. In this study, we sought to compare the dual and single drug-delivery system, to deliver the optimal dose of Resveratrol (RES) and Tretinoin (TTN) into the in vitro matured oocyte and morula-compact stage embryonic cells. The formation of single (RES/TTN) and dual-drug (RES + TTN)-SLN were confirmed by Uv-vis spectrophotometery, dynamic light scattering (DLS), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) technologies. In two experiments, the oocytes/presumptive zygotes were cultured under various concentrations of the single (RES/TTN) and dual-drug (RES + TTN)-SLN. In vitro toxicity studies, including nuclear staining (Aceto-orcein and Hoechst 33342), H2DCFDA fluorescent staining, chemiluminescence assay, and quantitative reverse transcription-PCR (qRT-PCR) techniques, indicated an excellent oocyte/embryo internalization of RES and TTN. Moreover, when oocytes/embryos were treated with the lowest concentration of RES + TTN-SLN, antioxidants-related genes were upregulated, apoptotic-related genes were downregulated, and intra/extracellular ROS production was reduced. In vitro cytotoxicity studies also demonstrated that single/dual-encapsulation of RES or TTN were safe even at the highest concentration (10 and 5 µM) compared to the control group. To sum it up, both delivery systems of RES and TTN by SLN (dual or single encapsulation) can deliver the optimal dose of RES and TTN into the oocyte/embryo. Where the dual-delivery of RES and TTN even at the lowest concentration (0.25 µM + 0.1 µm) showed a synergistic anti-oxidative effect in oocyte/embryo with a better inhibition of intra/extra-cellular ROS production by an enhanced/controlled intracellular penetration.

The Anti-oxidative Effects of Encapsulated Cysteamine During Mice In Vitro Matured Oocyte/Morula-Compact Stage Embryo Culture Model: a Comparison of High-Efficiency Nanocarriers for Hydrophilic Drug Delivery-a Pilot Study.

Although it is well-recognized that antioxidant nano-encapsulation has many benefits such as minimizing side effects (e.g., high-dose toxicity), the most attention was paid to the hydrophobic antioxidant not hydrophilic. In this regard, we sought to compare two hydrophilic model nanocarriers to deliver the optimal dose of cystamine (Cys) into the in vitro matured oocyte and the first cleavage stages until morula-compact stage embryonic cells. The formation of Cys-loaded solid self-emulsifying lipid (Cys + SLN) and Cys-loaded chitosan shell (Cys-CS-NC) were confirmed by FT-IR and UV-Vis spectrophotometry, dynamic light scattering (DLS), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) technologies. In two experiments, the oocytes/presumptive zygotes were cultured under various concentrations of Cys-SLN and Cys-CS-NC. The results of nuclear staining (aceto-orcein and Hoechst 33342), H2DCFDA fluorescent staining, chemiluminescence test, and quantitative reverse transcription-PCR (qRT-PCR) technique as in vitro toxicity studies demonstrated that adding the lowest dose of Cys-encapsulated in both nanocarriers [Cys-SLN (5 µM) and Cys-CS-NC (10 µM)] to maturation or culture medium could accumulate a strong anti-oxidative effect in oocyte/embryo by controlled release and enhanced intracellular penetration of Cys. In comparison, Cys-SLN (5 µM) is more effective than Cys-CS-NC (10 µM) groups to improve the expression of antioxidant genes (SOD, CAT, GPx) or anti-apoptotic (BCL-2) gene and decreased apoptosis (BAX and caspase-3) or intra-/extracellular ROS levels. In a nutshell, both nanocarriers (CS-NC or SLN) can deliver the lowest dose of Cys into the oocyte/embryo, thus encouraging a better expansion of antioxidant genes and enhancing the development of in vitro oocyte/embryo.

The impact of endocrine disruptor chemicals on oocyte/embryo and clinical outcomes in IVF.

The negative impact of endocrine-disrupting pesticides on human fertility is now a key issue in reproductive health. There are much fewer literature data about the impact of pesticide exposure on women than on men and very few studies of women participating in an in vitro fertilization (IVF) programme. In the present review, we found that (1) various pesticides with an endocrine-disrupting action are associated with poor oocyte maturation and competency, embryonic defects and poor IVF outcomes, and (2) some pesticide compounds are linked to specific causes of female infertility, such as premature ovarian insufficiency, polycystic ovarian syndrome, and endometriosis. IVF participants living in agricultural regions should be informed about the fertility decline, low ongoing pregnancy rates, and elevated risk of miscarriage associated with exposure to high doses of pesticides.

Endometriosis and strategies of fertility preservation: a systematic review of the literature.

Endometriosis is a common chronic inflammatory disorder, often causing both pain and infertility. It is estimated that 25-50 % of patients undergoing fertility treatments have had endometriosis as it involves an impairment of the ovarian reserve. For these reasons, endometriosis has been highlighted as a condition that may require a fertility preservation procedure, while being benign in nature. The aim of this review is to summarize the current evidence on fertility preservation techniques for patients affected by endometriosis, focusing on the main characteristics of the different approaches. A systematic review of literature was performed by searching in the main electronic databases (MEDLINE, EMBASE, Web of Science, Scopus, ClinicalTrial.gov, OVID and Cochrane Library), from their inception to February 2020 for studies testing fertility preservation (FP) techniques. Only scientific publications in English were included. Risk of Bias Assessment was performed. Eight articles were included in the study: 3 case reports (one paper reporting 2 cases), 1 prospective longitudinal analysis with only 1 reported case with criteria for inclusion in the review, 2 retrospective observational studies and 2 retrospective cohort studies. The results of our analysis showed embryo (1 article), oocyte (4 articles) and ovarian tissue cryopreservation (3 articles) as preservation methods proposed in international literature for patients with endometriosis. Only few authors reported details on successive outcomes. After ovarian tissue transplantation, 1 pregnancy with IVF and 1 case of endocrine function/ovulation recovery were described; six of 16 endometriotic patients, who underwent embryo cryopreservation, experienced livebirths after successive embryo-transfer; in a large series of 485 patients performing oocyte cryopreservation, an overall cumulative live-birth rate/patient of 46.4 % (225 babies) was reported. Based on the risk of premature ovarian failure related to endometriosis, the offer of FP techniques to these patients has significantly increased, as well as the reported experiences in recent medical literature. However, further studies concerning risks, benefits, effectiveness and cost-effectiveness are needed.

The ZAR1 protein in cancer; from epigenetic silencing to functional characterisation and epigenetic therapy of tumour suppressors.

ZAR1, zygote arrest 1, is a zinc finger protein (C-terminus), which was initially identified in mouse oocytes. Later it was found that its expression is present in various human tissues e.g. lung and kidney. Interestingly, it was observed that in various tumour types the ZAR1 transcript is missing due to hypermethylation of its CpG island promoter, but not ZAR2. Since methylation of the ZAR1 promoter is described as a frequent event in tumourigenesis, ZAR1 could serve as a useful diagnostic marker in cancer screens. ZAR1 was described as a useful prognostic/diagnostic cancer marker for lung cancer, kidney cancer, melanoma and possibly liver carcinoma. Furthermore, ZAR1 was reactivated as a tumour suppressor by epigenetic therapy using CRISPR-dCas9 method. This method holds the potential to precisely target not only ZAR1 and reactivate tumour suppressors in a tailored cancer therapy. ZAR1 is highly conserved amongst vertebrates, especially its zinc finger, which is the relevant domain for its protein and RNA binding ability. ZAR1 is implicated in various cellular mechanisms including regulation of oocyte/embryo development, cell cycle control and mRNA binding, though little was known about the underlying mechanisms. ZAR1 was reported to regulate and activate translation through the binding to TCS translation control sequences in the 3'UTRs of its target mRNA the kinase WEE1. ZAR1 has a tumour suppressing function by inhibiting cell cycle progression. Here we review the current literature on ZAR1 focusing on structural, functional and epigenetic aspects. Characterising the cellular mechanisms that regulate the signalling pathways ZAR1 is involved in, could lead to a deeper understanding of tumour development and, furthermore, to new strategies in cancer treatment.

Fertility preservation in Turkey: a global look for nationwide strategy development

As the reproductive technology advanced along with the improved outcome in cancer treatment demands implementing new fertility preservation, developing algorithms on fertility preservation requires tailoring for each society. Here, the authors attempt to modify the current medical literature on fertility preservation for the Turkish population. A PubMed search was conducted using the search term fertility preservation. Initially, 280 items of literature were accessed. In the second evaluation, 126 articles were examined and 154 items were discarded due to the low quality of the literature. In the final round, only 68 publications that were the most relevant were found eligible for inclusion in this review article. In order to develop a more systematic national guideline, forming a multidisciplinary approach to create a web-based network would be the first step. Both physicians and patients will have open access to the information. This database should be linked to an international consortium to stay integrated and open for updating. The aim of this review was to evaluate the relationship between the current situation in our country and the developments in the world in light of the literature, and to establish infrastructure for the development of future approaches in our country.

Impact of a gestational exposure to diesel exhaust on offspring gonadal development: experimental study in the rabbit.

The aim of the present work was to address experimentally the possible impact of exposure to air pollution during gestation on the differentiation and function of the gonads of the offspring using a rabbit model. Rabbits were exposed daily to diluted diesel exhaust gas or filtered air from the 3rd until the 27th day of gestation, during which time germ cells migrate in genital ridges and divide, and fetal sex is determined. Offspring gonads were collected shortly before birth (28th day of gestation) or after puberty (7.5 months after birth). The structure of the gonads was analyzed by histological and immunohistological methods. Serum concentrations of testosterone and anti-Müllerian hormone were determined using ELISA. The morphology and the endocrine function of the gonads collected just at the arrest of the exposure were similar in polluted and control animals in both sexes. No differences were observed as well in gonads collected after puberty. Sperm was collected at the head of the epididymis in adults. Sperm motility and DNA fragmentation were measured. Among all parameters analyzed, only the sperm DNA fragmentation rate was increased three-fold in exposed males. Mechanisms responsible for these modifications and their physiological consequences are to be further clarified.

Premature menopause - Meeting the needs.

Premature menopause is characterized by amenorrhea, elevated gonadotropin levels and sex steroid deficiency occurring in women below 40 years of age. Heterogeneity of premature ovarian failure is reflected by various causes, both spontaneous and iatrogenic. X chromosomal abnormalities remain the main cause of primary ovarian insufficiency. Women with primary ovarian insufficiency present with amenorrhea, irregular cycles, signs of estrogen deficiency and infertility. They are at increased risk for non-communicable diseases such as, cardiovascular disease, Alzheimer's disease, and osteoporosis. Management includes use of menopausal hormone therapy till the age of natural menopause and customized as per the needs. In women having fertility issues, IVF with donor oocytes remains the treatment of choice with the best results. With advances in assisted reproductive technologies, cryopreservation of oocytes/embryos should be utilized, whenever impending primary ovarian insufficiency is anticipated either due to therapy or biological reasons.

Descarga de la maduración final ovocitaria con gonadotropina coriónica humana o un agonista de la GnRH en donantes de ovocitos: análisis retrospectivo de cohortes

El protocolo de antagonistas de la GnRH (antGnRH) ha permitido el uso del agonista de la GnRH (aGnRH) como inductor de la descarga ovulatoria (DO), en reemplazo de la gonadotropina coriónica humana (hCG). Una ventaja importante con respecto al uso del aGnRH en la DO es que disminuye significativamente el riesgo del síndrome de hiperestimulación ovárica (SHEO), debido a su efecto luteolítico, de importancia en casos de donantes de ovocitos. Objetivos: Comparar la proporción de ovocitos obtenidos, calidad ovocitaria/embrionaria, resultado reproductivo y la incidencia de SHEO en ciclos de donación de ovocitos consecutivos, donde la DO fue con hCG o aGnRH. Adicionalmente, se comparó los ciclos con diagnóstico genético preimplantacional. Diseño: Estudio retrospectivo de cohortes. Institución: Grupo PRANOR, Clínica Concebir, Lima, Perú. Participantes: Donantes y receptoras de ovocitos. Intervenciones: Se utilizó un protocolo antGnRH en 43 donantes de ovocitos para 203 ciclos de estimulación consecutiva y DO con hCG (n=127) o aGnRH (n=76) basados en una decisión clínica y del conteo folicular. Los ovocitos fueron asignados, parte de ellos, a un banco de ovocitos (vitrificación) o a 303 ciclos de receptoras de ovocitos. Principales medidas de resultados: Ovocitos aspirados, tasa de fecundación, calidad embrionaria, formación a blastocisto. Resultados: Se observó diferencias significativas en el número de ovocitos aspirados a favor del grupo aGnRH sobre el grupo hCG (24,09±12,24 versus 18,69±8,93, p=0,002, respectivamente). No hubo diferencias significativas en el resultado global de proporción de ovocitos maduros, tasa de fecundación, calidad embrionaria, formación a blastocisto entre los grupos comparados. La proporción de embriones cromosómicamente normales fueron similares entre los grupos aGnRH (35,2%) y hCG (34,53%), mientras que las tasas globales de embarazo (64,49% versus 62,59%), implantación (45,41% versus 42,38%) y aborto (8,7% versus 8,7%) fueron comparables. La incidencia de SHEO fue reducida significativamente en el grupo de DO con aGnRH (0%), en comparación al grupo hCG (18,9%, p<0,0001). Conclusiones: La DO con el aGnRH es un método seguro y eficaz para ser aplicado en pacientes con alto riesgo de SHEO, como las donantes de ovocitos, eliminando por completo el SHEO. La calidad embrionaria y resultados reproductivos fueron idénticos entre los grupos de DO con aGnRH y hCG.

The GnRH antagonists (GnRHant) protocol has allowed the use of GnRH agonist (GnRHa) as inductor of ovulatory discharge (OD) replacing human chorionic gonadotropin (hCG). An important advantage of GnRHa use in OD is that it significantly decreases the risk of ovarian hyperstimulation syndrome (OHSS) due to its luteolytic effect, important in cases of oocyte donors. Objectives: To compare proportion of oocytes obtained, oocyte/embryo quality, reproductive result and OHSS incidence in consecutive oocyte donation cycles when OD was done with hCG or GnRHa. Cycles with preimplantation genetic diagnosis were compared. Design: Cohort retrospective study. Setting: Grupo PRANOR, Clinica Concebir, Lima, Peru. Participants: Oocyte donors and receptors. Interventions: A GnRHant protocol was used in 43 oocyte donors for 203 consecutive stimulation cycles and OD with hCG (n=127) or GnRHa (n=76) based in clinical decision and follicular count. Oocytes were assigned either to an oocyte bank (vitrification) or to 303 cycles of oocyte receptors. Main outcome measures: Oocytes aspired, fecundation rate, embryo quality, blastocyst formation. Results: There were significant differences in number of oocytes aspired in favor of the GnRHa group with regards to the hCG group (24.09±12.24 versus 18.69±8.93, p=0.002 respectively). There were no significant differences among the groups compared in the global result of mature oocytes proportion, fecundation rate, embryo qulity, blastocyst formation. The proportion of chromosomically normal embryos was similar between GnRHa (35.2%) and hCG (34.53%) groups, and pregnancy (64.49% versus 62.59%), implantation (45.41% versus 42.38%) and abortion (8.7% versus 8.7%) global rates were comparable. OHSS incidence reduced significantly in the OD group with GnRHa (0%) compared with the hCG group (18.9%, p<0.0001). Conclusions: OD with GnRHa is a safe and efficacious method to use in patients with high risk of OHSS such as oocyte donors, as it completely eliminates OHSS. Embryo quality and reproductive results were identical between OD groups with GnRHa and hCG.