26 research outputs found
Differential expression of progesterone receptor isoforms related to PGR +331g/a polymorphism in endometriosis: A case-control study
Background: Endometriosis are defined as a progesterone-resistance disease. Two progesterone receptor (PR) isoforms, namely PR-A and PR-B, mediate the special effects of progesterone. One of the most effective polymorphism in the promoter region of PGR is the +331G/A.Objective: The differential expression level of PR isoforms due to +331G/A polymorphism may be able to influence the function of progesterone and reduce the susceptibility of endometriosis.Materials and Methods: This analytic, case-control study was carried out at Royan Institute, Tehran, Iran. Whole-blood samples were collected from 98 infertile women undergoing laparoscopy for endometriosis and 102 healthy fertile women. After DNA extraction, genotype frequencies were determined by polymerase chain reactionrestriction fragment length polymorphism. Then, RNA was extracted from the selectedeutopic tissue samples of endometriosis patients. Analysis of PR-A and PR-B mRNA expressions were performed using Real-time polymerase chain reaction.Results: The frequency distribution of GG, GA genotypes in +331G/A polymorphism was 98.04%, 1.96% in the patients and 97.96%, 2.04% in the control groups, respectively (p = 0.968). Although our data did not show any significant association with +331G/A in the patient and control groups, we were able to demonstrate significantly higher expression level of PR-B and no significant lower expression level of PR-A isoforms in patients by favoring GA to GG genotypes (p = 0.017, p = 0.731, respectively).Conclusion: Our findings show that patients with GA genotypes had a higher expression level of PR-B compared to patients with GG genotypes
Valsalva maneuver using a Handmade Device in Supraventricular Tachycardia Reversion; a Quasi Experimental Study
Introduction: The use of vagal nerve stimulation is identified as a proper treatment option in patients with stable supraventricular tachycardia (SVT). This study aimed to assess the success of Valsalva maneuver via a handmade device in reversion of SVT. Methods: In this quasi experimental study, using a handmade device, vagus nerve stimulation was performed for SVT patients presenting to emergency department or cardiac intervention unit and the success rate and its related factors were assessed. Results: 100 patients with the mean age of 53.05 ± 13.70 years were studied (67% female). 12 (12%) cases were unable to do the maneuver. Out of the 88 (88.0%) patients who could perform the maneuver, 75 (85.2%) cases were unsuccessful. Dysrhythmia was controlled in 6 (6.8%) cases on the first attempt and in 7 (8.0%) cases on the second one (14.8% total success rate). 12 of the 13 cases (92.3%) with successful maneuver had history of SVT (p = 0.031). There was not any significant association between success rate and sex (p = 0.084), age (p = 0.744), or other medical histories (p ≥ 0.05). Conclusion: Based on the results of the present study, the success rate of Valsalva maneuver with the mentioned handmade device was calculated to be 14.8%. The only independent related factor of successful reversion was SVT history.
Increased expression of stemness genes Rex-1, Oct-4, Nanog, and Sox-2 in women with ovarian endometriosis versus normal endometrium: A case-control study
Background: Endometriosis is a common, chronic inflammatory disease which is defined as an overgrowth of endometrial tissue outside the uterine cavity. The etiology of this disease is complex and multifactorial but there is a strong evidence that supports the presence of endometrial stem cells and their possible involvement in endometriosis.
Objective: In this study, we analyzed the mRNA expression of REX-1 stemness gene and reconsidered three other stemness genes SOX-2, NANOG, OCT-4 in women with endometriosis compared to normal endometrium.
Materials and Methods: Ten ectopic and ten eutopic tissue samples along with 23 normal endometrium specimens were recruited in this study. The expression levels of OCT-4, NANOG, SOX-2, and REX-1 genes were evaluated by the quantitative real-time polymerase chain reaction.
Results: The transcription levels of OCT-4, NANOG, and SOX-2 mRNA were significantly increased in ectopic lesions compared with eutopic and control group (p = 0.041, p = 0.035, p = 0.048), although the REX-1 mRNA increase was not significant between endometriosis and control groups. Also, there were differences in the expression level of these genes in normal endometrium during the menstrual cycles (p = 0.031, p = 0.047, p = 0.031).
Conclusion: Based on our data, we confirm the dynamic role of stemness genes in proliferation and growth of normal endometrium during the menstrual cycle and conclude that differential expression n levels of these genes may contribute to the pathophysiology of endometriosis.
Key words: Endometriosis, Stemness genes
TIMPs Expression as A Maternal Cell Free Plasma Biomarker of Severe Preeclampsia: A Case-Control Study
Objective:
Preeclampsia (PE) is a pregnancy related disorder with prevalence of 6-7%. Insufficient trophoblasticinvasion leads to incomplete remodeling of spiral arteries and consequent decrease in feto-placental perfusion. Alteredplacental expression of tissue inhibitors of matrix metalloproteinase (TIMPs) is considered to be involved in this processwhile the balance between matrix metalloproteinases (MMPs) and TIMPs contributes to remodeling of the placentaand uterine arteries by degradation and refurbishing of extracellular matrix (ECM). Therefore, TIMPs, fetal expressionpattern was evaluated with the aim of its potential to be used as a determinant for the (early) detection of PE.
Materials and Methods:
In this case-control study, cell free fetal RNA (cffRNA) released by placenta into the maternalblood was used to determine expression patterns of TIMP1, 2, 3 and 4 in the severe preeclamptic women in comparisonwith the normal pregnant women. Whole blood from 20 preeclamptic and 20 normal pregnant women in their 28-32weeks of gestational age was collected. The second control group consisted of 20 normal pregnant women in either 14or 28 weeks of gestation (each 10). cffRNA was extracted from plasma and real-time polymerase chain reaction (PCR)was done to determine the expression levels of TIMP1, 2, 3 and 4 genes.
Results:
Statistical analysis of the results showed significant higher expression of TIMP1-4 in the preeclamptic womenin comparison with the control group (P=0.029, 0.037, 0.037 and 0.049, respectively). Also, an increased level of TIMPsexpression was observed by comparing 14 to 28 weeks of gestational age in the normal pregnant women in the secondcontrol group.
Conclusion:
An increased cffRNA expression level of TIMPs may be correlated with the intensity of placental vasculardefect and may be used as a determinant of complicated pregnancies with severe preeclampsia
Differential Incorporation of β-actin as A Component of RNA Polymerase II into Regulatory Regions of Stemness/Differentiation Genes in Retinoic Acid-Induced Differentiated Human Embryonic Carcinoma Cells
Objective
Nuclear actin is involved in transcription regulation by recruitment of histone modifiers and chromatin remodelers to the regulatory regions of active genes. In recent years, further attention has been focused on the role of actin as a nuclear protein in transcriptional processes. In the current study, the epigenetic role of nuclear actin on transcription regulation of two stemness (OCT4 and NANOG) and two differentiation) NESTIN and PAX6) marker genes was evaluated in a human embryonal carcinoma cell line (NT2) before and after differentiation induction.
Materials and Methods
In this experimental study, differentiation of embryonal cells was induced by retinoic acid (RA), and quantitative real-time polymerase chain reaction (PCR) was used to evaluate differential expression of marker genes before and 3 days after RA- induced differentiation. Chromatin immunoprecipitation (ChIP) coupled with real-time PCR was then undertaken to monitor the incorporation of β-actin, as a functional component of RNA polymerase II, in the regulatory regions of marker genes.
Results
Data showed significant change in nuclear actin incorporation into the promoter regions of NESTIN and PAX6 after RA-induction.
Conclusion
We emphasize the dynamic functional role of nuclear actin in differentiation of embryonal cells and its role as a subunit of RNA polymerase II
Epigenetic Regulation of Osteogenic and Chondrogenic Differentiation of Mesenchymal Stem Cells in Culture
Management of mesenchymal stem cells (MSCs) capabilities to differentiate into osteogenic and chondrogenic lineages would be of utmost importance for their future use in difficult to treat cases of destroyed bone and cartilage. Thus, an understanding of the epigenetic mechanisms as important modulators of stem cell differentiation might be useful. Epigenetic mechanism refers to a process that regulates heritable and long-lasting alterations in gene expression without changing the DNA sequence. Such stable changes would be mediated by several mechanisms including DNA methylation and histone modifications. The involvement of epigenetic mechanisms during MSC bone and cartilage differentiation has been investigated during the past decade. The purpose of this review is to cover outstanding research works that have attempted to ascertain the underlying epigenetic changes of the nuclear genome during in vitro differentiation of MSCs into bone and cartilage cell lineages. Understanding such genomic alterations may assist scientists to develop and recognize reagents that are able to efficiently promote this cellular differentiation. Before summarizing the progress on epigenetic regulation of MSC bone and cartilage differentiation, a brief description will be given regarding in vitro conditions that favor MSC osteocytic and chondrocytic differentiation and the main mechanisms responsible for epigenetic regulation of differentiation
Expression level of chromodomain Y (CDY): potential marker for prediction of sperm recovery in non-obstructive azoospermia
Background: The availability of testis specific genes will be of help in choosing the most promising biomarkers for the detection of testicular sperm retrieval in patients with non-obstructive azoospermia (NOA). Testis specific chromodomain protein Y 1 (CDY1) is a histone acetyltransferase which concentrates in the round spermatid nucleus, where histone hyperacetylation occurs and causes the replacement of histones by the sperm-specific DNA packaging proteins, TNPs and PRMs.
Objective: The aim was to evaluate CDY1 gene as a marker for predicting of successful sperm retrieval in NOA patients.
Materials and Methods: This research was conducted on 29 patients with NOA who had undergone testicular sperm extraction (TESE) procedure. NOA patients were subdivided into patients with successful sperm retrieval (NOA+, n=12) and patients with unsuccessful sperm retrieval (NOA-, n=17). Relative expression of CDY1 gene and chromatin incorporation of CDY1 protein were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and ELISA assay, respectively.
Results: Quantification of mRNA relative expression and incorporation of CDY1 protein in chromatin showed significant lower expressions and protein levels of CDY1 in testis tissues of NOA- in comparison to NOA+ group.
Conclusion: The findings in this study demonstrated a correlation between the low levels of CDY1 function and unsuccessful sperm recovery in the testicular tissues of NOA- compared to NOA+ patients. Therefore, it can be reasonable to consider CDY1 as a potential biomarker for predicting the presence of spermatozoa, although the claim needs more samples to be confirmed
Differences in fatty acid profiles and desaturation indices of abdominal subcutaneous adipose tissue between pregnant women with and without PCOS
The objective was to determine the differences in fatty acid (FA) profiles in subcutaneous adipose tissue (AT) between pregnant women with polycystic ovary syndrome (PCOS) and those without PCOS. FA profiles of AT samples from 13 PCOS and 32 non-PCOS, all of whom underwent caesarean section were compared using gas chromatography. Age and BMI in the two groups were similar. Twenty-one FAs were detected and the total saturated FA percentage of experimental groups was similar. While the total monounsaturated FA (MUFA) (p < 0.0004) and desaturase index (18:1 cis-9/18:0; p < 0.03) were higher in PCOS women than non-PCOS women, total polyunsaturated FA (PUFA) was lower in PCOS than non-PCOS women (p < 0.004). Docosahexaenoic acid level of the two groups was similar while α-linolenic acid and eicosapentaenoic acid levels were significantly (p < 0.05) lower in PCOS. Total trans-FA, C18:1 t9 and C18:2t were lower in PCOS women (p < 0.05). These results indicate differences in desaturase index, MUFA and PUFA, especially n-3 FA in AT between age and BMI-matched pregnant PCOS and non-PCOS pregnant subjects. Further studies are warranted to replicate these findings and to investigate potential changes in these profiles in non-pregnant PCOS women
Epigenetic role of the nuclear factor NF-Y on ID gene family in endometrial tissues of women with endometriosis: a case control study
Abstract Background A predominant difference between endometrial and normal cells is higher proliferation rate in the former cells which is benign. The genes of inhibitor of differentiation (ID) family play a major role in cell proliferation regulation which might be targeted by the nuclear transcription factor Y (NF-Y) for subsequent epigenetic modifications through the CCAAT box regulatory region. The present study was designed to investigate the epigenetic role of NF-Y on ID gene family in endometrial tissue of patients with endometriosis. Materials & methods In this case-control study, 20 patients with endometriosis and 20 normal women were examined for the relative expression of the NF-YA, NF-YB, NF-YC and ID genes by real-time PCR during the proliferative phase. The occupancy of NF-Y on CCAAT box region of ID genes was investigated using chromatin immunoprecipitation (ChIP) followed by real-time PCR. Results The NF-YA was over-expressed in eutopic endometrium during the proliferative phase. Although the expression level of NF-YB and NF-YC were unchanged in eutopic samples, they were remarkably higher in ectopic group (P<0.05). The ID2 and ID3 genes were up-regulated in ectopic and eutopic tissues, however ID1 and ID4 genes were down-regulated in these samples (P<0.05). The ChIP analysis revealed significant enrichment of NF-Y on regulatory regions of ID2,3 genes in eutopic group, but reduced binding level of NF-Y to the ID1,3 promoters in ectopic specimens (P<0.05). Conclusion The ability of NF-Y to regulate ID genes via CCAAT box region suggests the possible role of NF-Y transcription factor in epigenetic changes in endometrial tissues which may open novel avenues in finding new therapeutic strategies