ABSTRACT
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.
ABSTRACT
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.
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: 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: Recurrent miscarriage, as the occurrence of two or more of pregnancy loss before the 20[th] wk, can occur for multiple causes. One of the causes of miscarriage may be a defect in the process of angiogenesis because the delivery of nutrients to the fetus is decreased and it may lead to miscarriage. Also, micro ribonucleic acids play an important role in the development of diseases. The microRNAs 16 and 21 are the most well-known angiogenesis-related miRNAs, which their gene targets are vascular endothelial growth factor-A and phosphatase and tensin homolog, respectively
Objective: To evaluate the changes in expression of microRNAs 16 and 21 and their association with the gene targets in women with unexplained RM
Materials and Methods: In this case-control study, blood samples were taken from 25 women with unexplained RM and 25 controls. After extraction of RNA, the relative expression of microRNAs and their gene targets was measured using real-time quantitative reverse transcription-PCR method
Results: Our findings showed that miR-21 expression was significantly decreased in both plasma and peripheral mononuclear cells [p=0.04 and p=0.02, respectively] and could be associated with the PTEN expression [p=0.03], however, there is no significant correlation between miR-16 and VEGF-A
Conclusion: One of the most remarkable results of this study is that miR-21 showed significant changes in both plasma and peripheral mononuclear cells, which can be related to the etiology and progression of RM
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
Transendothelial migration of leukocytes [diapedesis], have vital role in both the innate and adaptive immune responses. This process consists of overlapping steps, including: activation of leukocytes, formation of weak adhesions, and translocation along the endothelium followed by stronger adhesions resulting in transmigration of leukocytes. Although the surface molecules used by leukocytes and endothelial cells during diapedesis have been well characterized, the mechanisms by which they regulate this process is poorly understood. In this mini review we introduce some of the involved molecules and mechanisms that play roles in this process