Possible Harmful Effects of Smoking Hookah on Sperm DNA Fragmentation Index and Protamine Genes Expression in Normozoospermic Men.

Objectives: In recent years, smoking water pipes or hookah has increased among adolescents in most countries. Although there is evidence in support of the negative effects of this type of smoking on human health, such as the increased risk of lung disease, little is known about the potential effects of hookah smoking on the male reproductive system, especially on the molecular aspects of sperm. Patients and methods: This cross-sectional study examined sperm DNA fragmentation index, protamine 1 and 2 (PRM1 and PRM2) genes expression, and oxidant status in normozoospermic hookah smokers in comparison with non-smoker controls. Results: Our results showed significantly higher rates of DNA fragmentation, protamine deficiency, and abnormal chromatin condensation in the spermatozoa of hookah smokers (P < .0001). Also, protamine gene expression showed a remarkable decrease in hookah smokers (1.55 ± 2.54 and 0.33 ± 0.54) compared to the controls (3.49 ± 5.41 and 1.22 ± 1.96), although the reduction was not statistically significant (P = .155 and P = .066, respectively). Moreover, a significantly higher level of semen MDA was observed in the case group compared to the controls (0.39 ± 1.04 vs 0.15 ± 0.21; P = .013). Conclusion: According to our study, although hookah smoking does not have a significant effect on sperm parameters, it may have deleterious effects on DNA integrity, oxidative status, and nuclear protein levels of spermatozoa.

Proliferating cell nuclear antigen presentation, as a marker of folliculogenesis, in the transplanted ovarian tissue.

AIM: One of the most important ways to understand the ovarian biology is studding the initiation of primordial follicle development and subsequent folliculogenesis control. In this study, proliferating cell nuclear antigen (PCNA) presentation was used as a marker of follicular development in the thawed ovarian tissue (OT) following transplantation onto chick embryo chorioallantoic membrane (CAM) using two methods of freezing of slow freezing and vitrification. METHODS: Samples of OT from 10 patients were subjected to slow freezing and vitrification. After warming, CAM transplantation was done and PCNA proliferation index (PI; percent of PCNA-positive granulosa cells) was calculated for each follicle stage. Image J software was used to determine the mean staining intensity. RESULTS: PCNA was positive for granulosa cells and oocytes nuclei, but negative for ooplasm. There were no remarkable PCNA staining in the granulosa cells of primordial follicles, but increased significantly as follicle progression (p < 0.05). Proliferation rate was also insignificantly higher in the vitrified than slow freezing group, before and after transplantation (p < 0.05). Lower PCNA presentation index was observed after CAM transplantation (p < 0.05). The earliest stage of follicular recruitment took place in the transitional follicles, before squamous cells transform to cuboidal cells. CONCLUSION: PCNA showed that follicles had proliferation power after cryopreservation. Higher presentation after vitrification may indicate accelerated folliculogenesis in the thawed OT.

Vitrification has detrimental effects on maturation, viability, and subcellular quality of oocytes post IVM in cancerous women: An experimental study.

BACKGROUND: In vitro maturation (IVM) of immature oocytes retrieved from ovarian tissue has been considered as a valuable approach for fertility preservation in cancerous patients. OBJECTIVE: To evaluate the efficacy of vitrification on oocyte maturation, survival rates, as well as the subcellular oocyte quality post IVM. MATERIALS AND METHODS: The ovarian cortexes from 19 women with cervix and uterine malignancy aged 21-39 yr were collected. Cumulus-oocyte complexes were aspirated from all visible antral follicles. 102 immature oocytes were collected, and 43 oocytes were detected appropriately for IVM (control group). Also, 59 immature oocytes were vitrified, then matured in vitro (IVM) in two groups: with Growth/differentiation factor 9 (GDF9) (group 1) and without GDF9 (group 2) supplementation. Rates of oocytes viability, maturation, and survival along with meiotic spindle visualization and zona pellucida birefringence were assessed with Polyscope. RESULTS: The rate of maturation was significantly higher in controls (55.8%) compared to the other groups. Maturation rate was 23.3% in oocytes cultured in IVM medium enriched with GDF9, and 27.6% in those cultured in IVM medium lacking GDF9 (p = 0.86). Also, the meiotic spindle was present in 74.4% of control oocytes which was significantly higher than the other groups. The proportion of high zona pellucida birefringence was higher in the controls when compared with group 1 (51.2% vs. 23.3%, respectively, p = 0.04). CONCLUSION: Vitrification had a detrimental effect on oocyte maturation, viability as well as the subcellular quality of the oocytes after IVM in cancerous women.

Biological and physiological characteristics of human cumulus cells in adherent culture condition.

BACKGROUND: Cumulus cells, as oocyte nurse cells, provide a suitable microenvironment with growth factors and cellular interactions required for oocyte maturation. Thus, these cells may serve as a natural niche for in vitro studies of female germ cell development. Cumulus cells may help attain a better understanding of the causes of infertility in women and eventually improve the outcomes of cases that respond poorly to standard infertility treatment. OBJECTIVE: The aim of this study was to isolate, culture, and investigate the biological characteristics of human cumulus cells. MATERIALS AND METHODS: In this experimental study, cumulus cells were isolated, cultured, and characterized using reverse transcription-polymerase chain reaction analyses of specific genes including FOXL2, CYP19A1, FSHR, AMHR, and LHR. The presence of vimentin, a structural protein, was examined via immunofluorescent staining. Moreover, levels of anti-mullerian hormone (AMH) and progesterone secretion by cumulus cells were measured with ELISA after 2, 4, 12, 24, and 48 hr of culture. RESULTS: In adherent culture, human cumulus cells expressed specific genes and markers as well as secreted AMH and progesterone into the medium. CONCLUSION: Cumulus cells secrete AMH and progesterone in an adherent culture and might be applicable for in vitro maturation (IVM) and in vitro gametogenesis (IVG) studies.

The effect of immature oocytes quantity on the rates of oocytes maturity and morphology, fertilization, and embryo development in ICSI cycles.

PURPOSE: The goal was to evaluate the role of the number of retrieved immature oocytes on mature oocyte counts and morphology, and also the rates of fertilization and embryo development in ICSI cycles. METHODS: 101 ICSI cycles were included in this prospective evaluation. Patients were divided into 2 groups of A (≤ 2 immature oocytes) and B (> 2 immature oocytes). In sub-analysis, the impacts of the number of GV and MI oocytes were assessed on the rates of fertilization and embryo development. Also, correlations between the numbers of immature and mature oocytes, as well as maternal age between two groups were analyzed. Assessments of oocyte morphology, fertilization, embryo quality and development were done accordingly. RESULTS: There was no correlation between the immature oocytes quantity with the number of mature ones. There were insignificant differences for embryo development between two groups, but fertilization rate was higher in group A (P = 0.03). In sub-analysis, insignificant differences were observed between two groups of ≤ and >2 GV and MI oocytes for rates of fertilization and embryo development. Also, the rates of clinical pregnancy and delivery were insignificant between groups. The rate of morphologically abnormal oocytes had no significant difference between two groups, except for wide perivitelline space (PVS) which was higher in group A (P = 0.03). There was no significant difference for maternal age between two groups. CONCLUSIONS: In cases with few retrieved immature oocytes, rates of fertilization and incidence of wide PVS may increase, although immature oocytes may not have any negative impacts on early embryo development, or the rates on number of mature oocytes.