ABSTRACT
As numerical and structural defects in chromosomes are an inevitable consequence of IVF, Pre-implantation genetic diagnosis and screening [PGD/PGS] methods are used for detecting abnormalities in embryos before implantation to the uterus to increase the successful rate of IVF. Pre-implantation genetic diagnosis and screening approaches can be achieved by different techniques such as NGS, CGH and FISH. Among these approaches, FISH-based PGD/PGS is challenging in that it requires experience and skill to increase its facility and validity. Therefore, based on literature review and our experiences obtained from genetic laboratory of Yazd Reproductive Sciences Institute [Yazd, Iran], we were ditermined to discuss these challenges. After reviewing the available protocols and articles, we compared results of different methods for performing pre- and post-examination FISH process. Required samples in each section were obtained from embryo in cleavage or blastocyst stage. According to our team's experience, we recommend the cleavage stage biopsy and our modified fixation method. Also, we do not recommend more than two round hybridization on the same cell. Many studies have shown that FISH-based PGD is an efficient method for decreasing IVF failure in infertile patients. This paper introduces the best biopsy and fixation method and, includes some useful tips and tricks on type and number of probe, removing the cytoplasm, denaturation and hybridization, data evaluation and scoring criteria
ABSTRACT
Background: tumor protein p53 [TP53] is a tumor suppressor transcriptional regulator protein which plays a critical role in the spermatogenesis. One of the most important regulators of p53 is Murine double minute 2 [MDM2], which acts as a negative regulator of the p53 pathway. Based on the key role of p53 and MDM2 in germ cell apoptosis, polymorphisms that cause a change in their function might affect germ cell apoptosis and the risk of male infertility
Objective: this study was designed to examine associations of TP53 72 Arg>Pro [rs1042522], and MDM2 309 T>G [rs937283] polymorphisms with spermatogenetic failure in Iranian population
Materials and Methods: a case-control study was conducted with 150 nonobstructive azoospermia or severe oligozoospermia and 150 fertile controls. The two polymorphisms, 72 Arg>Pro in TP53 and 309 T>G in MDM2, were genotyped using PCR-RFLP and ARMS-PCR respectively
Results: our analyses revealed that the allele and genotype frequencies of the TP53 R72P polymorphism were not significantly different between the cases and controls [p=0.41, p=0.40 respectively]. Also, no significant differences were found in the allelic [p=0.46] and genotypic [p=0.78] distribution of MDM2 309 T>G polymorphism between patients and controls
Conclusion: the results of this study indicate that polymorphisms of TP53 and MDM2 genes are unlikely to contribute to the pathogenesis of male infertility with spermatogenetic failure
ABSTRACT
Background: the study of microRNA expression can be effective in the diagnosing and treating different diseases. miR-135a is one of the most important micro-ribonucleic acids involved in endometriosis. Among the genes that become the target of the miR-135a and are subjected to changes in the endometrium of patients with endometriosis is HOXA10 gene which is expressed in the endometrium in response to steroid hormones
Objective: the aim of this study was to evaluate the expression of miR-135a and its relationship with the level of HOXA10 gene expression in both endometrial ectopic and eutopic tissues in patients with endometriosis compared to the control samples
Materials and Methods: in this prospective case-control study, both case-eutopic and case-ectopic tissue samples were obtained from 17 women with endometriosis and the eutopic endometrial tissue was sampled from 17 women with normal endometrium as the control group. The gene's expression of miR-135a and HOXA10 were investigated using quantitative reverse transcription PCR [q-RT PCR]
Results: a significant decrease in the expression of HOXA10 gene was detected in case-eutopic during the luteal phase compared to the control samples [p=0.001], while in the case-ectopic, the expression of this gene was increased [p=0.681] compared to the control samples. In addition, the expression miR-135a in the luteal phase showed a remarkable increase in the case-eutopic endometrial tissue [p=0.026] as well as a significant decrease in the case-ectopic endometrial tissue compared to the control samples [p=0.008]
Conclusion: considering the inverse relations between the over-expression of miR-135a and the reduction of HOXA10, it seems that miR-135a may be applied as an endometrial diagnostic and therapeutic biomarker
ABSTRACT
Background: genetic factors are believed to play an important role in the etiology of polycystic ovarian syndrome [PCOS] which is the most common endocrinological disorder of women in their reproductive age. Androgen metabolism is impaired in PCOS and, thus, CYP19 gene which is involved in this pathway can be a candidate gene. Previous studies have shown a relationship between single nucleotide polymorphism [SNP] of CYP19 in hyperandrogenism and PCOS in some racial groups
Objective: this study was designed to elucidate the role of CYP19 gene in PCOS in Iran
Materials and Methods: in this case-control study, 70 PCOS women and 70 non-PCOS women as normal control were selected. Following the informed consent, 5 ml blood was taken from individuals and subsequently, genomic DNA was extracted by salting out method. Furthermore, a set of polymerase chain reaction restriction fragment length polymorphism [PCR-RFLP] was carried out using specific primers for SNP rs.2414096 followed by enzyme digestion, with HSP92II
Results: genotype frequencies of SNP rs. 2414096 in PCOS women were as follows: AA [14.4%], AG [44.3%], and GG [41.4%] while in normal group, genotypes were 24.3%, 52.8%, and 22.9%, respectively. Allele frequencies in PCOS group were 49.3% for A and 50.7% for G, whereas normal group had a different percentage of A [36.4%] and G [63.6%]. The calculations for both genotypic and allelic frequencies showed statistical significance difference
Conclusion: variants of SNP rs. 2414096 in CYP19 could play a role in the development of PCOS in Iranian women
ABSTRACT
Background: Teratoasthenozoospermia [TA] is a severe form of male infertility with no clear etiology
Objective: To compare the level of intracellular anion superoxide [O[2]-], heat shock protein A2 [HSPA2] and protamine deficiencies in ejaculated spermatozoa between teratoasthenozoospermic and normozoospermic men
Materials and Methods: In this case- control study, semen samples of 20 infertile men, with TA [with normal morphology lower than 4%_ and total motility lower than 40% ] as the case group and 20 normozoospermic fertile men as the control group were evaluated for intracellular O[2] - and HSPA2 by flow cytometry and protamine deficiency by Chromomycin A3 [CMA3] test
Results: The rate of CMA3+ spermatozoa in the case group was higher than controls [p=0.001]. The percentages of HSPA2[+] spermatozoa in the cases were significantly lower than controls [p=0.001]. Also, intracellular O[2] - levels in the case group were significantly higher than controls [p=0.001] and had positive correlations with sperm apoptosis [r=0.79, p=0.01] and CMA3 positive sperm [r=0.76, p=0.01], but negative correlations with normal morphology [r=-0.81, p=0.01] and motility [r=-0.81, p=0.01]. There was no significant correlation between intracellular O[2] - and HSPA2 in the case group [r=0.041, p=0.79]
Conclusion: We suggest that the increase in intracellular O[2] -, decrease in spermatozoa HSPA2[+], and high percentages of spermatozoa with immature chromatin might be considered as etiologies of infertility in TA patients
ABSTRACT
Background: Selection of the best embryo for transfer is very important in assisted reproductive technology [ART]. Using morphological assessment for this selection demonstrated that the correlation between embryo morphology and implantation potential is relatively weak. On the other hand, aneuploidy is a key genetic factor that can influence human reproductive success in ART
Objective: The aim of this lab trial study was to evaluate the incidence of aneuploidies in five chromosomes in the morphologically high-quality embryos from young patients undergoing ART for sex selection
Materials and Methods: A total of 97 high quality embryos from 23 women at the age of 37or younger years that had previously undergone preimplantation genetic screening for sex selection were included in this study. After washing, the slides of blastomeres from embryos of patients were reanalyzed by fluorescence in-situ hybridization for chromosomes 13, 18 and 21
Results: There was a significant rate of aneuploidy determination in the embryos using preimplantation genetic screening for both sex and three evaluated autosomal chromosomes compared to preimplantation genetic screening for only sex chromosomes [62.9% vs. 24.7%, p=0.000]. The most frequent detected chromosomal aneuploidy was trisomy or monosomy of chromosome 13
Conclusion: There is considerable numbers of chromosomal abnormalities in embryos generated in vitro which cause in vitro fertilization failure and it seems that morphological characterization of embryos is not a suitable method for choosing the embryos without these abnormalities
ABSTRACT
N-acetyl cysteine [NAC], as a nutritional supplement, is a greatly applied antioxidant in vivo and in vitro. NAC is a precursor of L-cysteine that results in glutathione elevation biosynthesis. It acts directly as a scavenger of free radicals, especially oxygen radicals. NAC is a powerful antioxidant. It is also recommended as a potential treatment option for different disorders resulted from generation of free oxygen radicals. Additionally, it is a protected and endured mucolytic drug that mellows tenacious mucous discharges. It has been used for treatment of various diseases in a direct action or in a combination with some other medications. This paper presents a review on various applications of NAC in treatment of several diseases
Subject(s)
Humans , Antioxidants/therapeutic use , Oxidative StressABSTRACT
Background: Radiotherapy has many side effects on fertilization in young women. Radiation can lead to ovarian failure in women who underwent abdomen or pelvic radiotherapy
Objective: This study helps us to investigate ovarian response of NMRI female mice to ovarian stimulating hormones [PMSG, HCG] after whole-body gamma irradiation
Materials and Methods: 45 pregnant mice were divided into two groups of control and experimental. The experimental group was classified into three sub-groups: Irradiation group [2 or 4Gy],Superovulation group [10 or 15IU],and superovulation and gamma-radiation group [2Gy and 10IU, 2Gy and 15IU, 4Gy and 10IU,4Gy and 15IU]. Female mice were killed and embryos were removed from oviduct .The number of embryos cells counted and the quality of them was evaluated in each group. Kruskal-Wallis test and Mann-Whitney test were used to analyze the data
Results: There was a significant difference in the number of 2-4 cells grade D embryos in 2Gy and 15IU group compared with control and 2Gy groups [p=0.01], and the number of embryos in 4Gy group was more than in 10IU and 15IU [p=0.03] and 2Gy and 15IU groups [p=0.01]. It was more significantly embryos in 4Gy and 15IU group compared to 2Gy and 15IU group [p=0.01].In addition There were no significant differences in the number of 2-4 cells grades A, B and C embryos and also number of 4-8 cells grades A, B and C, D embryos in groups
Conclusion: The concurrent use of ovulation stimulating hormones and gamma rays ameliorates this problem of drastic decrease in number of living embryos due to whole-body irradiation
ABSTRACT
Background: methamphetamine [MA] is a potent psychomotor stimulant with high abuse and addictive potential. MA is a neurotoxic drug which is widely abused by females of childbearing age, raising serious public health concerns in terms of exposure of the fetus to the drug. Neurotoxic effects of MA on adult are well known, such as dopaminergic nerve terminal degeneration and cell death in regions of brain in some doses
Objective: in the present study, we examined effect of prenatal MA exposure on mouse fetuses
Materials and Methods: in this study, forty 8-12 week-old NMRI female mice were used which were mated with male mice in serial days. When sperm plug was observed it was designated as gestational day [GD] 0. Pregnant mice were individually housed in plastic cages. Pregnant mice were divided into four groups: in first group 10 mg/kg /day MA, in second group 5 mg/kg /day MA and in third group saline were injected subcutaneously from GD 6 to GD 14, corresponding to organogenesis period, while fourth or control group were without injection. On GD 14 fetuses were removed and accomplished chromosome preparation from fetal liver. Then fetal were fixed in formalin for brain hematoxilin and eosine staining and TUNEL assay
Results: we observed morphological abnormality including exencephal fetus in 5mg/kg MA group and premature fetuses in 10 mg/kg MA group. Also brain histological study showed subarachnoid hemorrhage in fetal brain in both experimental groups. Fetal liver karyotyping analysis was normal in fetuses of all groups and TUNEL assay in fetal striatum did not show significant difference in number of apoptotic cells between groups
Conclusion: from our results, it could be concluded that chronic abuse of MA by pregnant females during organogenesis period can cause teratogenic effect and brain hemorrage in fetus