Reanalysis of discarded blastocysts for autosomal aneuploidy after sex selection in cleavage-stage embryos / 대한생식의학회지

Objective@#The goal of the present study was to investigate the rate of chromosomal aneuploidies in surplus embryos after sex determination at the cleavage stage. Then, the same chromosomal aneuploidies were evaluated in blastocysts after extended culture. @*Methods@#Sixty-eight surplus embryos were biopsied at the cleavage stage and incubated for an additional 3 days to allow them to reach the blastocyst stage. The embryos were reanalyzed via fluorescence in situ hybridization (FISH) to examine eight chromosomes (13, 15, 16, 18, 21, 22, X, and Y) in both cleavage-stage embryos and blastocysts. @*Results@#Although the total abnormality rate was lower in blastocysts (32.35%) than in cleavage-stage embryos (45.58%), the difference was not significant (p=0.113). However, when we restricted the analysis to autosomal abnormalities, we observed a significant difference in the abnormality rate between the cleavage-stage embryos (44.11%) and the blastocysts (17.64%, p=0.008). A higher rate of sex chromosomal abnormalities was also observed in cleavage-stage embryos (29.4%) than in blastocysts (14.70%, p=0.038). @*Conclusion@#The data indicated that embryo biopsy should be conducted at the blastocyst stage rather than the cleavage stage. The results also emphasized that examination of common chromosomal aneuploidies apart from sex selection cycles can be conducted in the blastocyst stage with the FISH method.

Reanalysis of discarded blastocysts for autosomal aneuploidy after sex selection in cleavage-stage embryos / 대한생식의학회지

Objective@#The goal of the present study was to investigate the rate of chromosomal aneuploidies in surplus embryos after sex determination at the cleavage stage. Then, the same chromosomal aneuploidies were evaluated in blastocysts after extended culture. @*Methods@#Sixty-eight surplus embryos were biopsied at the cleavage stage and incubated for an additional 3 days to allow them to reach the blastocyst stage. The embryos were reanalyzed via fluorescence in situ hybridization (FISH) to examine eight chromosomes (13, 15, 16, 18, 21, 22, X, and Y) in both cleavage-stage embryos and blastocysts. @*Results@#Although the total abnormality rate was lower in blastocysts (32.35%) than in cleavage-stage embryos (45.58%), the difference was not significant (p=0.113). However, when we restricted the analysis to autosomal abnormalities, we observed a significant difference in the abnormality rate between the cleavage-stage embryos (44.11%) and the blastocysts (17.64%, p=0.008). A higher rate of sex chromosomal abnormalities was also observed in cleavage-stage embryos (29.4%) than in blastocysts (14.70%, p=0.038). @*Conclusion@#The data indicated that embryo biopsy should be conducted at the blastocyst stage rather than the cleavage stage. The results also emphasized that examination of common chromosomal aneuploidies apart from sex selection cycles can be conducted in the blastocyst stage with the FISH method.

Tumor necrosis factor Alpha -308 G/A single nucleotide polymorphism and risk of sperm abnormalities in Iranian males

Background: Signaling molecules such as cytokines regulate spermatogenesis during the maturation of germ cells and sperm apoptosis. Tumor necrosis factor alpha [TNF alpha] is one of the most-documented cytokines that is involved in spermatogenesis. We investigated the association of the TNF alpha -308 G/A single nucleotide polymorphism with sperm abnormalities in Iranian males

Materials and Methods: This case-control study included 180 infertile men who referred to Yazd Research and Clinical Center for Infertility and 100 healthy normospermic controls. Infertile men were classified into four groups of azoospermia [n=91], oligospermia [n=26], teratospermia [n=30] and asthenoteratospermia [n=33]. After sperm analysis, DNA was extracted from blood and polymerase chain reaction-restriction fragment length polymorphism [PCR-RFLP] was carried out for the genotyping of TNF alpha-308 G/A

Results: The A allele was significantly associated with sperm abnormality in our population [[P<0.001, odds ratios [OR] 95% confidence interval [CI]=2.31]. In addition, the A allele was also associated with azoospermia [P<0.001, OR [95% CI]=2.484], oligospermia [P=0.005, OR [95% CI]=2.51] and teratospemia [P<0.001, OR [95% CI]=3.385] but not with asthenoteratospermia [P=0.623]

Conclusion: Our data suggest that this single nucleotide polymorphism [SNP] maybe associated with the risk of sperm abnormality in infertile men of Iranian origin

Effects of gamma radiation on fetal development in mice

Background: Many cancer patients receive radiotherapy which may lead to serious damages to the ovary storage and the matrix muscle state. Some of these patients may admit to infertility clinics for having pregnancy and on the other hand hormonal administration for superovulation induction is a routine procedure in assisted reproduction technology [ART] clinics

Objective: This study aimed to investigate fertility and fetuses of hormone treated super ovulated female mice who had received whole-body gamma irradiation before mating.Materials and Methods: Female mice were randomly categorized into a control group and 3 experimental groups including: Group I [Irradiation], Group II [Superovulation], and Group III [Superovulation and Irradiation]. In hormone treated groups, mice were injected with different doses of 59Tpregnant mare's serum gonadotropin59T [PMSG] followed with human chorionic gonadotropin [HCG]. Irradiation was done using a Co-60 gamma ray generator with doses of 2 and 4 Gy. Number of fetuses counted and the fetus's weight, head circumference, birth height, the number of live healthy fetuses, the number of fetuses with detected anomalies in the body, the sum of resorption and arrested fetuses were all recorded as outcome of treatments

Results: In the group I and group II, increased radiation and hormone dose led to a decrease in the number of survived fetuses [45 in 2 Gy vs. 29 in 4 Gy for irradiated group] as well as from 76 in 10 units into 48 in 15 units. In the group III, a higher dose of hormone in the presence of a 2 Gy irradiation boosted the slink rate; i.e. the number of aborted fetuses reached 21 cases while applying the dose of 15 Iu, whereas 6 cases of abortion were reported applying the hormone with a lower dose. Among different parameters studied, there was a significant difference in parameters of weight and height in the mouse fetuses [p=0.01]

Conclusion: The data indicated that use of ovarian stimulating hormones in mice that received pre mating gamma irradiation did not significantly increase the pregnancy rates

Haplotype analysis of BRCA1 intragenic markers in Iranian patients with familial breast and ovarian cancer

Background: Breast cancer is the most common malignancy in women. Breast Cancer Type 1 Susceptibility gene [BRCA1] is a tumor suppressor gene, involved in DNA damage repair and in 81% of the breast-ovarian cancer families were due to BRCA1. In some clinically investigated genes, the intragenic marker polymorphism is important and the screening of such mutations is faster by using short tandem repeat [STR] polymorphism. Individual polymorphism of STR is a good evidence for following inheritance of repeat polymorphism

Objective: The aim of this study was to evaluate three intragenic BRCA1 marker polymorphisms in families, which have two or more patients with breast/ovarian cancer in comparison to healthy women

Materials and Methods: A total of 107 breast and/or ovarian cancer patients and 93 unrelated healthy women with no clinical phenotype of any malignancy or familial cancer history constitute the study groups. Haplotyping analysis, at 3 intragenic BRCA1 microsatellite markers [D17S855, D17S1322 and D17S1323], were performed for all subject and control groups using labeled primers

Results: After fragment analysis, significance differences were observed as follows: two alleles of D17S855; allele 146 [p=0.02] and 150 [p=0.006], and two alleles of D17S1322, allele 121 [p=0.015] and 142 [p=0.043]. These differences were compared with control group. There was significance difference in 8 di/tri allelic haplotypes in present experimental subjects. Some haplotypes were observed to have approximately twice the relation risk for breast cancer

Conclusion: According to recent results, assessment of presence or absence of mentioned alleles in BRCA1 microsatellite can be used for prognosis in individuals, suspected of having or not having the breast cancer

Mutation analysis of exon1 of bone morphogenetic protein-15 gene in Iranian patients with polycystic ovarian syndrome

Background: with the prevalence of 6-10%, polycystic ovarian syndrome [PCOS] is considered the most common endocrinological disorder affecting women in their reproductive age. It has been suggested that genetic factors participate in the development of PCOS. Follicular development has been considered as one of the impaired processes in PCOS. Bone morphogenetic protein-15 [BMP-15] gene is a candidate gene in follicular development and its variants may play role in pathogenesis of PCOS

Objective: to investigate whether BMP-15 gene mutations are present in Iranian women with PCOS

Materials and Methods: in this cross-sectional study 5 ml venous blood samples was taken from 70 PCOS women referring to Afzalipour Hospital, Kerman University of Medical Sciences, Kerman, Iran, between January to December 2014. Genomic DNA was extracted from the blood sample by salting out method. Then a set of PCR reactions for exon1 of BMP-15 gene was performed using specific primers followed by genotyping with direct sequencing

Results: two different polymorphisms were found in the gene under study. In total 20 patients [28.6%] were heterozygote [C/G], and 2 patients [2.86%] were homozygous [G/G] for c.-9C>G in 5´UTR promoter region of BMP-15 gene [rs3810682]. In addition, in the coding region of exon1, three patients [4.3%] were heterozygote [G/A] for c.A308G [rs41308602]. Two PCOS patients [2.86%] appeared to have both c.-9C>G [C/G] and c.A308G [G/A] variants simultaneously

Conclusion: our research detected two polymorphisms of BMP-15 gene among PCOS patients, indicating that even though it cannot be concluded that variants of BMP-15 gene are the principal cause of polycystic ovarian syndrome; they could be involved in pathogenic process in development of PCOS

SULF 1 gene polymorphism, rs6990375 is in significant association with fetus failure in IVF technique

Sulfatase 1 [SULF1] function is to remove the 6-O-sulphate group from heparan sulfate. This action changes the binding sites of extracellular growth factors. SULF1 expression has been reported to be changed in angiogenesis. We hypothesized that single nucleotide polymorphisms [SNPs] of SULF1 would impact clinicopathologic characteristics. Study of SULF1 gene polymorphism with fetus failure in in vitro fertilization [IVF] technique. We studied one common [minor allele frequency >0.05] regulatory SNP, rs6990375, with polymerase chain reaction and restriction fragment length polymorphism method, in 53 infertile women with fetus failure in IVF technique and 53 women with at least one healthy child as controls. We found that rs6990375 is significantly associated with an early failure in IVF and frequency of G allele is high in women with fetus failure in IVF technique [p<0.001]. These findings suggest that SULF1genetic variations may play a role in IVF technique fetus failure. Further studies with large sample sizes on SULF1 SNPs may be useful in support of this claim

cellular uptake of antisense oligonucleotid of E6 mRNA into cervical cancer cells by DOPE-modified hydroxyapatite nanoparticles

Although several chemical and physical methods for gene delivery have been introduced, their cytotoxicity, non-specific immune responses and the lack of biodegradability remain the main issues. In this study, hydroxyapatite nanoparticles [NPs] and 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine [DOPE]-modified hydroxyapatite NPs was coated with antisense oligonucleotide of E6 mRNA, and their uptakes into the cervical cancer cell line were evaluated. Calcium nitrate and diammonium phosphate were used for the synthesis of the hydroxyapatite nanoparticle. Thus, they were coated with polyethylene glycol [PEG], DOPE and antisense oligonucleotide of E6 mRNA using a cross-linker. Then, hydroxyapatite NPs and DOPE-modified hydroxyapatite NPs were incubated 48 hours with cervical cancer cells and their uptakes were evaluated by fluorescent microscopy. The hydroxyapatite NPs had different shapes and some agglomeration with average size of 100 nm. The results showed DOPE-modified hydroxyapatite NPs had higher uptake than hydroxyapatite NPs [P<0.05]. Hydroxyapatite NPs conjugated with DOPE are a good choice for gene delivery and silencing of viral genes in cervical cancer cells, but their efficacy should be addressed more in future studies

luteinizing hormone beta-subunit exon 3 [Gly102Ser] gene mutation and ovarian responses to controlled ovarian hyperstimulation

Despite extensive progress in IVF techniques, one of the most difficult problems is the variability in the response to controlled ovarian hyperstimulation [COH]. Recent studies show the effects of individual genetic variability on COH outcome. To evaluate the correlation between LHbeta G1502A polymorphisms in exon 3 of the LH gene and ovarian response to COH. A total of 220 women treated with a long protocol for ovarian stimulation were studied. Three genotypes of GG, GA and AA were detected by polymerase chain reaction-restriction fragment length polymorphism [PCR-RFLP] analysis. In total, 34 [17%] patients were poor responders, 154 [77%] were normal responders and 12 [6%] were hyper responders. The most frequent genotype was GA [55.5%] whereas 44.5% of patients showed GG genotype and there was no patient with AA genotype. In total 54.5% of normal responders, 61.8% of poor responders and 50% of hyper responders showed GA genotype. Our results did not establish a significant relationship between this polymorphism and the ovarian response. Therefore it is still very difficult to use the genotype of patients for prediction of the ovarian response to stimulation

Effect of nano-zinc oxide on doxorubicin- induced oxidative stress and sperm disorders in adult male Wistar rats

Doxorubicin [DOX], an anthracycline antibiotic, is a widely used anticancer agent. In spite of its high antitumor efficacy, the use of DOX in clinical chemotherapy is limited due to diverse toxicities, including gonadotoxicity. We investigated the protective effect of nano-zinc oxide [nZnO] as an established antioxidant on DOX-induced testicular disorders. In this experimental study 24 adult male Wistar rats were divided into four groups including one control and three experimentals [6 rats per group]. They received saline [as control], DOX alone [6 mg/kg body weight, i.p.], nZnO alone [5 mg/kg body weight, i.p.], and nZnO followed by DOX. Animals were sacrificed 28 days after treatment and evaluations were made by sperm count and measuring sex hormone levels in plasma. Also total antioxidant power [TAP] and lipid peroxidation [LPO] in plasma were tested. Data was analyzed with SPSS-14 and one way ANOVA test. P<0.05 were considered to be statistically significant. In the DOX-exposed rats significant differences were found compared with the control group [p=0.001] in plasma total antioxidant power [TAP] [425.50 +/- 32.33 vs. 493.33 +/- 18.54 mmol/mL], Lipid peroxidation [LPO] [3.70 +/- 0.44 vs. 2.78 +/- 0.68 micromol/mL], plasma testosterone [3.38 +/- 0.69 vs. 5.40 +/- 0.89 ng/dl], LH [0.26 +/- 0.05 vs. 0.49 +/- 0.18 mlU/mL], sperm count [157.98 +/- 6.29 vs. 171.71 +/- 4.42x10[6]/mL] and DNA damage [11.51 +/- 3.45 vs. 6.04 +/- 2.83%]. Co-administration of nZnO significantly improved DOX-induced changes [p=0.013] in plasma TAP [471.83 +/- 14.51 mmol/mL], LPO [2.83 +/- 0.75 micro mol/mL], plasma testosterone [5.00 +/- 1.07 ng/dl], LH [0.52 +/- 0.08 mlU/mL], sperm count [169.13 +/- 5.01x10[[6]/mL] and DNA damage [7.00 +/- 1.67%]. At the dose designed in the present investigation cytoprotective role of nano-zinc oxide through its antioxidant potential is illuminated in DOX-induced male gonadotoxicity.

Multiplex PCR screening of Y-chromosome microdeletions in azoospermic ICSI candidate men

It has been hypothesized that Y-q microdeletion can account for significant proportion of infertility in men. There are three nonoverlapping regions referred to as the "azoozpermia factors" AZFa, AZFb, and AZFc from proximal to distal part of Y-q. These have been defined as spermatogenesis loci, this region deletions have been shown to be involved in male azoospermic or severe oligoozospermic infertility. Evaluation the rate of Y-chromosome microdeletions in infertile men. In this case-control study, 25 azoospermic infertile men candidate for intracytoplasmic sperm injection [ICSI] were selected as case group. For control group, 25 normoozoospemric men were selected. All cases and controls had normal 46XY karyotype. DNA extraction and molecular analysis were done on blood samples. Multiplex-PCR method was done to identify the presence of microdeletion in AZFa, AZFb or AZFc loci. Eight STS primers that include two controls were selected to determine Y-chromosome microdeletions. 20% [5/25] of all patients have at least one microdeletion in more than one region of AZF loci. Totally 17 microdeletions was observed, one case had deletions in three AZF regions, and 4 cases had deletions in two AZF regions. The rate of deletions was 42% [7/17] for AZFc, 35% [6/17] for AZFa and 23% [4/17] for AZFb. The molecular DNA analysis could help us to know the real cause of infertility and can give good information for good decision for example in men whit microdeletions who want to undertake ICSI procedure the deletions will be passed to their son

point mutations of mitochondrial tRNA threonine and proline in idiopathic repeated pregnancy loss

Mitochondrial transfer RNAs [tRNA] genes are essential components of protein biosynthesis. These genes are hotspots for mutations. These mutations are associated with a wide spectrum of human disease. Many genetic factors are known in assessment of repeated pregnancy loss [RPL]. The aim of this study was analysis of tRNA[Thr] and tRNA[Pro] in women with RPL. The nucleotide variations of threonine and proline were investigated in 96 women with idiopathic repeated pregnancy loss. The related mitochondrial area was amplified using a polymerase chain reaction [PCR]. The PCR products were demonstrated by 2% agarose gel electrophoresis, and all the positive samples were purified and verified by an automated DNA sequencing method. The sequence analysis revealed 4 mutations in tRNA[Thr]. These mutations were A15907G in 2 cases [2.08%], A15924G in 3 cases [3.12%], G15928A in 10 cases [10.42%] as the most common mutations and G15930A in 3 cases [3.12%] as a novel mutation. Also, the result of tRNA[pro] sequencing showed the T15972C mutation in 1 woman [1.04%] as a novel mutation. These tRNAs mutations can alter their steady state level and affect the structure of tRNAs. It results in protein synthesis defects and, in turn, mitochondrial dysfunction. The mutations of these genes may help in the assessment of RPL. Further study of an expanded series of these tRNA mutants is recommended to describe their etiologic role in idiopathic RPL

Detection of chalmydia trachomatis infection in female partners of infertile couples

The prevalence of infertility is about 10-15% among the couple in reproductive age. Several factors can affect on fertility ability in men and women. Chlamydia is non-motile gram negative obligatory interacellular pathogenic organism. It can be caused infections in females as cervicitis, urethritis, endometritis, pelvic inflammatory disease also prostatitis and epidydidimitis in male as well. The aim of this survey is to mention the frequency of infection with Chlamydia infection in infertile female who were treated in Yazd research and Clinical Centre for Infertility. A questionnaire contains some demographic information and a clinical feature related to the infection was completed for each infertile women. Specimen of vaginal discharge was collected by well trained nurses using sterile cotton swap from 91 women. Elisa test was done on blood serum. DNA extraction for Chlamydia was carried out using low salt method and PCR was done using MOMP and plasmid primers. DNA sequencing was performed on two PCR products using Chromas LITE ver. 2.01 and analyzed by BLAST. Of 91 blood samples collected in this survey, none of those was positive by ELISA. Also there was not positive by PCR. Four PCR products showed a questionable band which were not in range of Chlamydia. The products were performed DNA sequencing and there were not related to any micro-organism. However, it is well known that Chlamydia has an infection role in infertility. Nevertheless, there was not evidence of this organism in these infertile patients. It is necessary to design such survey in large population of infertile patients especially on infertile women with tubal infertility and their husbands as well

Public knowledge and perceptions of biotechnology and genetically modified organisms in Iran

Biotechnology offers a variety of potential environmental, social and economic benefits but, it is the center of extreme public and political debate at present. A major field of discussion in biotechnology are genetically modified [GM] organisms. New medical, political, ethical and religious discussions arise over the production and consumption of these organisms. Many surveys have been conducted in industrialized countries to investigate the public perceptions regarding the risks and benefits of biotechnology, while in developing countries hardly any studies have been done so far. The present descriptive study intends to contribute to a better understanding of public attitudes toward biotechnology and GM foods in Iran. To assess the public's knowledge and opinions on the dangers or opportunities of genetic modification, 300 university students as a sample of educated community and 300 individuals with no university's education as a sample of ordinary people were asked to complete a specially designed questionnaire on the risks and benefits of biotechnology and GM foods. This article summarizes the current situation with regard to biotechnology, with a particular focus on GM foods and discusses the results of the survey. Our results indicated that public's knowledge about biotechnology is low in Iran and more efforts are needed to improve their understanding of different aspects of biotechnology

relationship between seasonal variability and pregnancy rates in women undergoing assisted reproductive technique

Backgrownd: Studies in regions with seasonal climatic variations have revealed a correlation between human natural conception and birth rates. Holidays and other cultural activities probably have influence on conception, but the ambient temperature and emotional influences on the female hormones related to fertility may play an important part in the seasonal variation in conception

Objectives: The aim of study was to determine the relationship between the success rate of Assisted Reproductive Technique [ART] treatment cycles and temperature in different seasons

Materials and Methods: A retrospective study on all individuals undergoing assisted ART at our institution was performed during June 2000 to June 2001. The study population represented 258 IVF-ET cycles and 821 ICSI treatment cycles. Different variables were analyzed using jj test

Results: In IVF treatment cycles, conception was more common from early spring [March to June]. This decreased from spring, with the minimum in fall, 22% and 14%, respectably. A significant seasonal variability in the number of eggs, embryo transferred and sperm motility was not demonstrated [p>0.05], but sperm count was significantly higher in spring than any other season [72 +/- 4 xl0[6] and 52 +/- 7xl0[6], respectively]

Conclusion: The seasonal changes should be taken into account together with other factors when evaluating infertility data