Investigating the effect of Nigella sativa on the testicular function of first-generation offspring of mice treated with titanium oxide nanoparticles.

Objective: Nanoparticles include primary particles with at least one of their dimensions being less than 100 nm. The goal of this research was to determine the possible protective role of Nigella sativa (NS) against toxic effects mediated by titanium oxide nanoparticle (TNP). Materials and Methods: 30 adult mice (10 males and 20 females) were used. After mating, the pregnant female mice were randomly divided into 4 study groups (n=5 mice in each group). From the 13th day of gestation until delivery, the mice were given TNP and NS. After delivery, 10 newborn male mice were selected from each group and kept under standard conditions until puberty according to the previous grouping (4 groups). The epididymis of each mouse was removed and the sperm was collected for the evaluation of in vitro fertilization and testis for histopathology and spermatogenesis of in vitro fertilization of first-generation mice. Results: No significant difference was observed between the NS group and the control group (p>0.05). In the TNP, a degree of epithelial lysis and a significant decrease in sperm motility was observed (p<0.05) compared with the control group. In the TNP and NS group, NS had an ameliorating effect on TNP-induced testicular germ cell damage (p<0.05). Conclusion: In the present study, it was found that NS had no destructive effect on the germinal epithelium. However, NS had an ameliorating effect on TNP-induced testicular germ cell damage in mice.

Assessment of Expression Levels and Localization Patterns of Phospholipase C zeta in Different Grades of HOST in Human Sperm.

BACKGROUND: Phospholipase C zeta (PLC-ζ) deficiency in sperm can underlie oocyte activation failure after intracytoplasmic sperm injection (ICSI). The aim of this study was to determine PLC-ζ expression and location in individual spermatozoa in each host score so that a hypo-osmotic swelling test (HOST) may be used to help routine sperm selection for ICSI. MATERIALS AND METHODS: In this experimental study, fresh semen samples were randomly obtained from 30 men who were referred to the Andrology Unit of the Infertility Center. Samples were processed by density gradient centrifugation (DGC) and exposed to hypotonic conditions. Seven different tail patterns, classified from 'a' to 'g' can be detected according to World Health Organization (WHO) criteria. Then, the PLC-ζ protein localization pattern was assessed by quantitative Immunofluorescence in individual sperm Host grades. Moreover, the sperm content of PLC-ζ protein was evaluated by flow cytometry correlated with semen analysis parameters. RESULTS: In the present study, quantitive immunofluorescence analysis indicated that sperm from different host grades exhibited seven localization patterns of PLC-ζ of acrosomal (A); equatorial (EQ), and postacrosomal (PA) patterns. A+EQ=acrosomal and equatorial, A+PA=acrosomal and post-acrosomal, EQ+PA=equatorial and post-crosomal, and A+EQ+PA. The sperm from HOST grade 'd' exhibited significantly higher PLC-ζ (A+PA) and (A+EQ+PA) staining compared to sperm from other grades (P=0.006). The sperm from grade 'd' exhibited higher PLC-ζ (EQ+PA) compared with other grades (P=0.001). However, grade 'd' was not significantly different from 'c' (P=0.087). Analysis of the combined results confirmed that there was a clear reduction in PLC-ζ immunofluorescence in Host grades 'a', 'f' and 'g' sperms. CONCLUSION: Our data suggest that HOST may represent a useful diagnostic tool for the selection of sperms exhibiting a higher level of PLC-ζ expression.

Fresh or Frozen Embryo Transfer in The Antagonist In Vitro Fertilization Cycles: A Retrospective Cohort Study.

BACKGROUND: Gonadotropin-releasing hormone antagonist (GnRH-ant), widely adopted protocol, is more in line with the physiological processes, and induces a shorter and more cost-effective ovarian stimulation. In order to assess the success rate of embryo transferring (ET) in the antagonist in vitro fertilization (IVF) cycles, we compared the fresh ET with the frozen ET outcomes. MATERIALS AND METHODS: In this retrospective cohort study, one hundred five cases of ET of the infertility clinic of the Besat hospital (Kurdistan, Iran) between March 2014 to March 2020 that were treated with antagonist cycle (both fresh and frozen) were analyzed. The difference between the two groups in baseline data and reproductive outcomes were evaluated using Independent sample t test, Mann-Whitney U test, Chi-squared test, and Fisher's exact test in SPSS software (version 22). RESULTS: Out of 105 cases, 48 and 57 were in the fresh and frozen ET groups, respectively. The participants age was 35.75 ± 4.9 Y. In the fresh ET group, and 33.98 ± 5.1 Y in the frozen ET group. The percentage of chemical pregnancy was 12 (25%) in the fresh ET group and 15 (26.3%) in the frozen ET group (P=0.8); Clinical pregnancy rate was 11 (22.9%) in the fresh ET group and 11 (19.3%) in the frozen ET group (P=0.6); the rate of abortion in the fresh ET group was 3 (6.3%, P=0.2), and in the frozen ET group was 8 (14%, P=0.2); and the live birth rate was 9 (18.8%) in the fresh ET group, in comparison with 7 (12.3%) in the frozen ET group (P=0.3). CONCLUSION: Not statistically significant, the percentage of chemical pregnancy and abortion were higher in the frozen ET group. The percentage of clinical pregnancy and live birth were higher in the fresh ET group.

Effects of The Mitochondrial Genome on Germ Cell Fertility: A Review of The Literature.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enters cells through angiotensin converting enzyme 2 (ACE2), which expression of its gene increases during pregnancy that is resulted in an enhanced level of the ACE2 enzyme. It might enhance the risk of SARS-CoV-2 infection and its complications in the pregnant women. Although, pregnancy hypertensive disorders and severe infection with SARS-CoV-2 are correlated with high comorbidity, these two entities should be discriminated from each other. Also, there is a concern about the risk of preeclampsia and consequently severe coronavirus disease 2019 (COVID-19) development in the pregnant women. So, to answer these questions, in the present review the literature was surveyed. It seems there is higher severity of COVID-19 among pregnant women than non-pregnant women and more adverse pregnancy outcomes among pregnant women infected with SARS-CoV-2. In addition, an association between COVID-19 with preeclampsia and the role of preeclampsia and gestational hypertension as risk factors for SARS-CoV-2 infection and its complications is suggested. However, infection of the placenta and the SARS-CoV-2 vertical transmission is rare. Various mechanisms could explain the role of COVID-19 in the risk of preeclampsia and association between preeclampsia and COVID-19. Suggested mechanisms are included decreased ACE2 activity and imbalance between Ang II and Ang-(1-7) in preeclampsia, association of both of severe forms of COVID-19 and pregnancy hypertensive disorders with comorbidity, and interaction between immune system, inflammatory cytokines and the renin angiotensin aldosterone system and its contribution to the hypertension pathogenesis. It is concluded that preeclampsia and gestational hypertension might be risk factors for SARS-CoV-2 infection and its complications.Infertility is one of the major problems faced in medicine. There are numerous factors that play a role in infertility. For example, numerous studies mention the impact of the quantity and quality of mitochondria in sexual gametes. This is a narrative review of the effects of the mitochondrial genome on fertility. We searched the PubMed, Science Direct, SID, Google Scholar, and Scopus databases for articles related to "Fertility, Infertility, Miscarriage, Mitochondria, Sperm, mtDNA, Oocytes" and other synonymous keywords from 2000 to 2020. The mitochondrial genome affects infertility in both male and female gametes; in sperm, it mainly releases free radicals. In the oocyte, a mutation in this genome can affect the amount of energy required after fertilisation, leading to gestation failure. In both cases, infertile cells have substantially less mitochondrial DNA (mtDNA) copies. The effects of mtDNA on gamete fertility occur via changes in oxidative phosphorylation and cellular energy production. Also, a reduction in the number of mtDNA copies is directly associated with sex cell infertility. Therefore, evaluation of the mitochondrial genome can be an excellent diagnostic option for couples who have children with neonatal disorders, infertile couples who seek assisted reproductive treatment, and those in whom assisted reproductive techniques have failed.

In vitro effects of pentoxifylline and coenzyme Q10 on the sperm of oligoasthenoteratozoospermia patients.

The effect of in-vitro sperm incubation with Pentoxifylline (PTX) and Coenzyme Q10 (CoQ10) in Oligoasthenoteratozoospermia (OAT) patients was evaluated. Semen samples were obtained from men with Normozoospermia and men with OAT. Motile sperm from the two groups were subdivided into four subgroups: (i) without incubation with PTX + CoQ10; (ii) incubation with PTX; (iii) Incubation with CoQ10; and (iv) incubation with a combination of PTX + CoQ10. Then, sperm parameters, chromatin, DNA and membrane integrity, protamine deficiency, apoptosis, mitochondrial activity, sperm chromatin dispersion test (SCD), hypo-osmotic swelling test (HOS), chromomycin A3 (CMA3), Terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL), and diaminobenzidine (DAB) assays were evaluated, respectively. Sperm incubated with CoQ10 and a combination of CoQ10 and PTX resulted in a significant increase in the sperm parameters. Also, a significant decrease was noted with a combination of PTX and CoQ10 in normal men. There was a significant difference between CoQ10 treated and CoQ10 + PTX treated groups in comparison with the OAT group in the percentage of the DNA fragmentation, sperm apoptosis, AB+, HOS test + and sperm mitochondrial activity. Incubated sperm with CoQ10, PTX, and in combination with each other can improve sperm parameters in OAT patients.

Effect of sperm selection methods on ICSI outcomes in patients with oligoteratzoospermia.

OBJECTIVE: Sperm selection without - or with a low level of - protamine deficiency and DNA fragmentation is a remarkable indicator to increase the success rate of ICSI outcomes. The aim of this study was to compare sperm selection methods in the elimination of sperm with protamine deficiency and DNA fragmentation and their effects on ICSI Outcomes in oligoteratzoospermia patients. METHODS: Semen samples were obtained from oligoteratozoospermia patients undergoing ICSI. Sperm selection was conducted using Zona Pellucida (ZP) binding, Hyaluronic Acid (HA) binding, and conventional PVP methods. SCD assay and CMA3 staining were used for the detection of sperm protamine deficiency and DNA fragmentation. Good quality of the embryo, blastocyst formation, chemical, and clinical pregnancy rates among studied groups was evaluated and compared. RESULTS: Our results indicated the percentage of sperm DNA fragmentation and protamine deficiency were lower significantly in the HA- and ZP-bound sperm. Although no significant differences were observed in the fertilization rate among studied methods, good quality of cleavage embryo rates were increased using ZP and HA methods versus the conventional PVP method. However, there were no significant differences in cleavage and embryo quality between the HA compared to the ZP method. Blastocyst formation, chemical and clinical pregnancy rates increased in the HA method. CONCLUSIONS: Overall, the HA method for sperm selection due to high sensitivity in selecting sperm with a low level of DNA fragmentation and protamine deficiency is a very useful method to increase the success rate of ICSI outcomes in oligoteratozoospermia patients.

The effect of platelet-rich plasma on the achievement of pregnancy during frozen embryo transfer in women with a history of failed implantation.

OBJECTIVE: The aim of this study was to evaluate the effect of platelet-rich plasma (PRP) on the rate of implantation and pregnancy in women with repeated failed implantation during frozen embryo transfer. METHODS: This study was conducted on 50 infertile women candidates (who were referred to the Infertility Treatment Center of Besat Hospital in Sanandaj) with a history of failed implantation for the purpose of frozen embryo transfer. The participants were randomly divided into two groups (n = 25). In the first group (control), the intrauterine infusion of 0.5 ml of Ringer serum was done 48 h before embryo transfer. In the second group (treatment), the intrauterine infusion of 0.5 ml of PRP was performed 48 h before embryo transfer. RESULTS: In this study, there was no significant difference between the two groups in the rate of chemical and clinical pregnancy. The rate of chemical pregnancy was 28% in the treatment group and 36% in the control group, while the rate of clinical pregnancy was 28% in the treatment group and 24% in the control group. CONCLUSION: The intrauterine infusion of PRP before frozen embryo transfer in infertile women with a history of failed implantation will not make any significant effect on the result of pregnancy.

Association of Factor V Leiden and Prothrombin G20210A Polymorphisms in Women with Recurrent Pregnancy Loss in Isfahan Province, Iran.

BACKGROUND: Maternal thrombophilia has been identified as a risk factor for recurrent pregnancy loss (RPL). The aim of this study was to investigate the association between prothrombin G20210A and factor V Leiden (FVL) polymorphisms in women with RPL and a control group of parous women in Isfahan province of Iran. METHODS: We studied 250 women with idiopathic RPL and 116 control cases. Prothrombin and FVL different genotypes were determined using polymerase chain reaction and reverse hybridization technique. RESULTS: The frequencies of heterozygous mutation prothrombin G20210A were 6% and 0.9%, respectively (P = 0.025), in cases compared to the control group. The frequencies of homozygous mutation prothrombin G20210A were 0.4% and 0%, respectively, in cases compared to controls (P = 0.02). The prothrombin mutation was significantly higher in cases compared to the control group (odds ratio 8.81; 95% confidence interval: 1.16-66.62). There was no significant difference between the FVL mutation and pregnancy loss. CONCLUSIONS: The results indicated a significant higher frequency of prothrombin G20210A in women with RPL in comparison with controls. Our data suggest that the prothrombin G20210A mutation, but not the FVL mutation, may be an unrecognized cause of RPL in our population.

Comparison of mitochondrial-related transcriptional levels of TFAM, NRF1 and MT-CO1 genes in single human oocytes at various stages of the oocyte maturation.

BACKGROUND: The aim of the current study was to assess the mRNA levels of two mitochondria-related genes, including nuclear-encoded NRF1 (nuclear respiratory factor 1), mitochondrial transcription factor A (TFAM), and mitochondrial-encoded cytochrome c oxidase subunit 1 (MT-CO1) genes in various stages of the human oocyte maturation. METHODS: Oocytes were obtained from nine infertile women with male factor undergoing in vitro fertilization (IVF)/intra-cytoplasmic sperm injection protocol. Mitochondrial-related mRNA levels were performed by single-cell TaqMan real-time PCR. RESULTS: the expression level of the target genes was low at the germinal vesicle stage (P>0.05). Although the mRNA level of NRF1gene remained stable in metaphase I, the mRNA level of TFAM and MT-CO1 increased significantly (P<0.05).In metaphase II, the expression level of all genes increased compared to metaphase I (P<0.05). CONCLUSION: The overexpression levels of NRF1, TFAM, and MT-CO1 genes are related to the oocyte maturation. Therefore, the current study could be used clinically to improve the success rate of IVF.

Methylenetetrahydrofolate Reductase C677T and A1298C Polymorphism in Iranian Women With Idiopathic Recurrent Pregnancy Losses.

BACKGROUND: Recurrent pregnancy loss (RPL) is a serious problem for pregnancy. There is evidence that vascular complications play a principal role in RPL. Methylenetetrahydrofolate reductase (MTHFR) is a key enzyme in folate metabolism. Polymorphisms (C677T, A1298C) of MTHFR gene are associated with decreased MTHFR activity. OBJECTIVES: The aim of this study was to determine the association between MTHFR polymorphisms (C677T and A1298C) and recurrent pregnancy loss (RPL) in Iranian women. MATERIALS AND METHODS: In this case-control study, blood samples were obtained from patients who had three or more consecutive pregnancy losses before the 22(nd) week of pregnancy (n = 204). The control group consisted of 116 age-matched women with at least one alive child and without any history of pregnancy loss or other gestational complications (n = 116). Following DNA extraction, samples were tested for MTHFR C677T and A1298C polymorphisms using the reverse hybridization method. RESULTS: The prevalence of 677TT mutation was 8.8% (18/204) in the patient group and 8.6% (10/116) in the control group (P = 0.434). The prevalence of 1298CC mutation was 12.3 % (25/204) in the patient group and 8% (9/116) in the control group (P = 0.155). Investigation of the distributions of various genotypes of MTHFR C677T and A1298C did not indicate a significant difference between patients with RPL and healthy control subjects. CONCLUSIONS: The results suggest that MTHFR mutations might not be associated with RPL in the examined population.