Frequency of the rs 14035 polymorphism of RAN gen in recurrent pregnancy loss: A case-control study

Background: Genetic factors could account for recurrent pregnancy loss (RPL). The RAN gene is a member of the ”large RAS family” and a small GTPase that is essential for the translocation of Ribonucleic acid (RNA) and proteins through the nuclear pore. Mutation in the RAN constitutive gene could stop DNA synthesis and alter the expression of genes in the uterus, likely playing a role in recurrent miscarriage. Objective: The aim was to investigate the frequency of RAN (rs 14035) polymorphism in women with RPL compared with women without abortion history. Materials and Methods: In this case-control study, 100 women with at least two consecutive miscarriages before the 20th wk of gestation and having spouses with karyotype and normal sperm parameters as the case group and 100 women with no history of abortion and having at least one successful pregnancy and normal delivery as the control group. The groups were age matched (20-40 yr). The rs 14035 polymorphism of RAN gene was investigated by Polymerase Chain Reaction- Restriction Fragment Length poly morphism technique and the frequency of which was compared between the two groups. Results: The frequency of TT, TC, and CC genotypes of RAN gene polymorphism in the case group were 9%, 40%, and 51%, respectively, and in the control group were 11%, 38%, and 51%, respectively. There was no significant difference in the genotypes between two groups (p = 0.882). Conclusion: According to our results, it seems that RAN polymorphism (rs 14035) is not associated with the risk of RPL in this study population. Key words: RAN gene, Repeated abortion, Polymorphism, PCR-RFLP

The effect of the human cumulus cells-conditioned medium on in vitro maturation of mouse oocyte: An experimental study

Background: To increase the results of infertility treatment, many efforts have been made to improve the treatment methods. As assisted reproductive technology is mainly using cell culture methods, one of the approaches to improve this technology is conditioned medium from different sources. It is desirable to apply in vitro maturation (IVM) and use oocytes from normal cycles instead of stimulating ovulation. Objective: To investigate the effect of human cumulus cell condition medium (hCCCM) on the IVM of immature mouse oocytes and morphology. Materials and Methods: In this experimental study, 240 germinal vesile oocytes were collected from four-six wk-old mice after 48 hr of 5IU pregnant mare serum gonadotropin (PMSG) injection and cultured in hCCCM (test group, n = 120) and DMEM + 20% FBS (control group, n = 120). The IVM rates and changes in perivitelline space (PVS) and shape were investigated at 8, 16, and 24 hr following the culture. The mature (MII) oocytes were subjected to in vitro fertilization (IVF) and the fertilization rate was assessed in three days. Results: A significant difference was observed between the maturation rates in the hCCCM and control groups (24.16% vs 0%; p = 0.001), as well as morphologic changes between the two groups (p = 0.04, p = 0.05). The development rate for MII oocytes attained from IVM in the hCCCM group was 27.58% (2-cell) and 6.89% (4-cell). Data displayed that hCCCM is an effective medium for oocytes maturation compared to the control medium. Conclusion: hCCCM supports oocyte in vitro growth and maturation. Moreover, hCCCM changes the oocyte shape and size of perivitelline space. Key words: Germinal vesicle, Cumulus cell, Conditioned medium, In vitro fertilization, In vitro maturation, Oocyte

The Effect of The Conditioned Medium from Human Embryonic Stem Cells on Mouse Oocytes In Vitro Maturation

Objective: Some reports have indicated that conditioned medium from growing mouse embryonic stem cells (ESCs)provides a supportive condition for small follicles growing, oocyte maturation, and following embryo growth. The aim ofthis study is assessing in vitro maturation (IVM) and consequent in vitro fertilization (IVF) outcome of immature mouseoocytes using human embryonic stem cells conditioned medium (HESCM).Materials and Methods: In this experimental study, 240 germinal vesicle (GV) oocytes were took from NMRI femalemice, aged 4-6 weeks, 48 hours before injection of 5 IU pregnant mare serum gonadotropin (PMSG). 120 GV oocyteswithout cumulus cells were cultured in each of the groups. 120 GV were cultured in HESCM as test groups and also120 GV cultured in human embryonic stem cells medium (HESM) as control groups. After evaluating the metaphase II(MII) oocyte maturation rate at 8, 16 and 24 hours, the MII oocytes subsequently were fertilized in vitro and the two-cellembryo development rate was recorded at days 1, 2, and 3. Statistical analysis was performed by using the generalizedestimating equations (GEE) method that calculated their rate ratio.Results: Our data indicated there are significant differences between the maturation rates in HESCM and HESM(P=0.004), also the two-cell embryo development was significant between two culture media (P=0.00).Conclusion: Similar to some other studies, the secretome of the HESCM showed a significant impact on the IVMoutcomes in mice

Exploring relationships between GSTP1 Ile105Val polymorphism and endometriosis risk in an Iranian population

Introdution: Endometriosis is one of the most common gynecologic disorders and shows significantly elevated prevalence in industrial regions. Additionally, a possible genetic predisposition is assumed for the disease. Glutathione S-transferases (GSTs) are enzymes participated in the metabolism of many human disease-causing mutagens, carcinogens and environmental pollutants. A functionally significant A to G transition in GSTP1 gene causes substitution of in the codon 105 can influence the enzyme activity. The aim of the present study was to investigate association of GSTP1 polymorphism and endometriosis in women from central and southern Iran. Methods: In this case-control study, after obtaining informed consent, samples were obtained from 101 endometriosis patients and 126 healthy controls. Genomic DNA was isolated from peripheral blood cells and genotyping was performed using polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) analysis. Data were compared in both groups by using Pearson chi-square and Hardy-Weinberg equilibrium tests. Results: Results showed no significant association between GSTP1 Ile105Val Polymorphism and endometriosis susceptibility (P = 0.370). Frequencies of the AA, AG and GG genotype of GSTP1 gene polymorphism in the patients were 42.6%, 47.5% and 9.9%, while the frequencies in the controls were 49.2%, 45.2% and 5.6%, respectively. Conclusion: According to our study, GSTP1 Ile105Val polymorphism appears to be not associated with the risk of endometriosis in the studied population. However, additional studies, especially with larger sample size are needed to validate these findings. &nbsp

WNT7A (rs104893832) polymorphism increases the risk of recurrent spontaneous abortion in Iranian women

BACKGROUND Recurrent spontaneous abortion is defined as the occurrence of three or more clinical miscarriages in one woman. Several factors, including genetics and environmental factors, are involved in this kind of infertility, in which WNT7A (rs104893832) polymorphism plays a major role. The aim of the present study was to determine the association between a common polymorphism of WNT7A (rs104893832) with recurrent spontaneous abortion in females. METHODS In the present case-control study, the WNT7A (rs104893832) polymorphism was investigated in 70 women with recurrent spontaneous abortion as cases and 100 women with at least one child and no history of infertility or abortion as controls. Polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) was used to investigate the WNT7A (rs104893832) polymorphism in both case and control groups. The data were subsequently analyzed using the chi-square and logistic regression tests by SPSS software (version 18.0). RESULTS A significant association was found between the WNT7A (rs104893832) polymorphism and recurrent spontaneous abortion (OR=25.00, 95% CI=5.52-157.09; p<0.0001). Our finding showed that G allele frequency in women with recurrent spontaneous abortion was significantly different compared to the control group. (OR=6.42, 95% CI=2.82-15.16; p<0.0001).Therefore, genetic variation in WNT7A (rs104893832) polymorphism may play a role in recurrent spontaneous abortion. Conclusion This study revealed that WNT7A (rs104893832) polymorphism increased the risk of recurrent spontaneous abortion. Knowledge of these mutations and polymorphisms can provide an insight into the prognosis for individual patients. Therefore, further studies are necessary to establish the association of WNT7A (rs104893832) polymorphism with recurrent spontaneous abortion in a larger population

No association of GSTM1 null polymorphism with endometriosis in women from central and southern Iran

Background: Endometriosis is one of the most common gynecologic disorders. It is a complex trait and both genetic and environmental factors have been implicated in its pathogenesis. There is growing evidence indicating that exposure to environmental contaminants is a risk factor for endometriosis. Glutathione-S-Transferase M1 (GSTM1) is one of the genes involved in detoxification of endogenous and exogenous compounds. Objective: Several studies have indicated an association between GSTM1 null mutation and endometriosis. In this study, the possible association between the GSTM1 gene null genotype and susceptibility to endometriosis in woman from central and southern Iran was investigated. Materials and Methods: One hundred and one unrelated premenopausal women with endometriosis and 142 unrelated healthy premenopausal women without endometriosis were enrolled in the study. Genomic DNA was extracted from Peripheral blood in all subjects. GSTM1 null genotyping was performed by polymerase chain reaction (PCR). Results: There was no significant difference between frequencies of GSTM1 null genotype in case and control groups (50.5% Vs. 52.1%, p=0.804). Furthermore, this genotype was not associated with severity of endometriosis in our sample (p=0.77). Conclusion: further studies involving gene-environment and gene-gene interactions, particularly combination of GSTM1 and other GST gene family polymorphisms are needed

The Effect of Zinc Supplementation on Expressed Levels of Peroxisome Proliferator-Activated Receptor Gamma and Glucose Transporter Type 1 Genes in Newborns of Women with Gestational Diabetes Mellitus

The current study was designed to determine the beneficial effects of zinc supplementation on expressed levels of peroxisome proliferator-activated receptor gamma (PPAR-γ) and glucose transporter type 1 (GLUT1) genes in newborns of women with gestational diabetes mellitus (GDM). This randomized, double-blind, placebo-controlled clinical trial was performed among 40 women with GDM. Patients were randomly allocated to intake either 233 mg zinc gluconate (containing 30 mg zinc) (n = 20) or a placebo (n = 20) for 6 weeks. PPAR-γ and GLUT1 mRNA levels were quantified in umbilical cord blood of newborns of women with GDM. After 6 weeks of intervention, the change in serum zinc levels was greater in women consuming zinc than in the placebo group (+11.1 ± 13.4 vs. −4.8 ± 17.3 mg/dL, P = 0.002). Quantitative results of RT-PCR demonstrated that compared with the placebo, zinc supplementation resulted in a significant increase of expressed levels of PPAR-γ mRNA (P < 0.001) and GLUT1 mRNA (P < 0.001) in umbilical cord blood of newborns of women with GDM. Taken together, the current study demonstrated that zinc supplementation for 6 weeks among GDM women increased the mRNA levels of PPAR-γ and GLUT1 in their newborns compared with the placebo group

Exome sequencing utility in defining the genetic landscape of hearing loss and novel-gene discovery in Iran

Hearing loss (HL) is one of the most common sensory defects affecting more than 466 million individuals worldwide. It is clinically and genetically heterogeneous with over 120 genes causing non-syndromic HL identified to date. Here, we performed exome sequencing (ES) on a cohort of Iranian families with no disease-causing variants in known deafness-associated genes after screening with a targeted gene panel. We identified likely causal variants in 20 out of 71 families screened. Fifteen families segregated variants in known deafness-associated genes. Eight families segregated variants in novel candidate genes for HL: DBH, TOP3A, COX18, USP31, TCF19, SCP2, TENM1, and CARMIL1. In the three of these families, intrafamilial locus heterogeneity was observed with variants in both known and novel candidate genes. In aggregate, we were able to identify the underlying genetic cause of HL in nearly 30% of our study cohort using ES. This study corroborates the observation that high-throughput DNA sequencing in populations with high rates of consanguineous marriages represents a more appropriate strategy to elucidate the genetic etiology of heterogeneous conditions such as HL