Biochemical Genetics (00062928)62(6)pp. 4317-4334
Papillary thyroid carcinoma (PTC), the most prevalent cancer of the thyroid, is more common in women than in men. To uncover the expression profile of FOXE1 gene in PTC tumor etiology. Microarray and RNA sequencing data on PTC in humans were analyzed. Eleven PTC tumor tissue samples and their neighboring normal tissue samples were collected. RT-qPCR was performed. Data normality, ROC construction, and logistic regression analysis were conducted. PTC tumors, normal tissues surrounding tumors, patients of different sexes and ages, metastasizing tumors, and tumor variants were assessed for FOXE1 expression. Eleven PTC tissues were obtained from seven women and four men. Among the PTC subtypes, there were two FV-PTCs, four C-PTCs, one microcarcinoma, and four tissues with an unknown subtype. FOXE1 gene expression was significantly increased in PTC tumors with dimensions less than 10 mm (relative expression = 14.437, p = 0.050). A significant increase in FOXE1 gene expression was observed in the normal tissue adjacent to the tumor, which was less than 10 mm in size, compared to the normal tissue adjacent to the tumor, which was larger than 10 mm (relative expression = 41.760, p = 0.0001). Females diagnosed with PTC showed a significant reduction in FOXE1 mRNA levels compared to their male counterparts (relative expression = 0.081, p = 0.042). In contrast to adjacent normal tissue, there was a significant reduction in FOXE1 gene expression in FV-PTC (relative expression = 0.044 and p = 0.0001). PTC tumors under 10mm had higher FOXE1 gene expression than larger tumors; normal tissue adjacent to smaller tumors also had higher FOXE1 expression. Females with PTC, regardless of their subtype, expressed less FOXE1 mRNA than males. FV-PTC tissues exhibited lower expression of FOXE1 mRNA than their adjacent normal tissues. © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2024.
Plant Cell, Tissue and Organ Culture (15735044)156(2)
Salinity stress is a major environmental factor that negatively impacts plant growth and agricultural productivity. Ethylene, a crucial plant hormone, plays a vital role in regulating various physiological processes in response to environmental stresses, including salinity stress. In this study, we investigated the effect of two ethylene inhibitors, pyrazinamide (PZA) and AgNO3, on the expression of key ethylene synthesis and signaling genes in tomato plants (Lycopersicon esculentum L.) under salinity stress in vitro conditions. Tomato seedlings were subjected to salinity stress by supplementing the growth medium with NaCl, and different concentrations of PZA and AgNO3 were applied. The expression patterns of ACS3, ACO1, EIN2, and EIL1 genes, involved in ethylene biosynthesis and signaling, were analyzed using real-time quantitative PCR (qPCR). Under salinity stress, the expression of ACS3 and ACO1 genes was upregulated, indicating increased ethylene production. However, the application of PZA and AgNO3 treatments effectively suppressed ACS3 and ACO1 gene expression by 23% and 14%, respectively. Similarly, the ethylene signaling genes, EIN2 and EIL1, exhibited increased expression under salinity stress, but their expression was significantly reduced by 60% to 88% with PZA and AgNO3 treatments. The study shows that PZA and AgNO3 inhibitors effectively improve stress tolerance in tomato plants under salinity stress. These results offer insights into ethylene-mediated responses and potential strategies for developing stress-resilient crop varieties. Further research is needed to explore interactions between inhibitors and the ethylene signaling pathway and validate their practical application in agriculture. © 2024, The Author(s), under exclusive licence to Springer Nature B.V.
Brazilian Journal of Otorhinolaryngology (18088694)90(4)
Objective: Nodular goiter may increase the risk of thyroid cancer, but the genetic factors contributing to nodular goiter are not well understood. There is an overexpression of H19 lncRNA in goiter tissue and its target remains unknown. In this study, we attempted to identify a new target for H19 in the context of goiter development. Methods: Using interaction energy calculations, the interaction between NKX2-1 mRNA and H19 lncRNA was examined. Putative microRNAs were found at the H19 lncRNA target site with the highest affinity for NKX2-1. RNAseq data was analyzed to determine the tissue specificity of gene expression. Samples were taken from 18 goiter and 18 normal tissues during thyroidectomy. The expression of NKX2-1 was determined by RT-qPCR using specific primers. Results: The interaction between NKX2-1 and H19 was characterized by six local base-pairing connections, with a maximum energy of −20.56 kcal/moL. Specifically, the sequence that displayed the highest affinity for binding with H19 overlapped with the binding site of has-miR-1827 to NKX2-1. It was found that NKX2-1 is exclusively co-expressed with H19 in normal thyroid tissue. As compared to adjacent normal tissues, nodular goiter tissues have a significant overexpression of NKX2-1 (relative expression = 1.195, p = 0.038). Conclusion: NKX2-1 has been identified as the putative target of H19 lncRNA, which is overexpressed in nodular goiter tissues significantly. Level of Evidence: 4. © 2024 Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial
Biologia (13369563)78(10)pp. 2717-2721
This correction stands to correct the original article, which due to a processing error was published without its figures. The publisher regrets the error and provided herein are the missing figures. Effect of melatonin and NaCl on fresh weight (A), dry weight (B) and root length (C) of alfalfa root. Values are means (± SD) of fifteen replicates. Different letters indicate significant differences (P < 0.05) based on Duncan’s multiple range test. Melatonin concentrations of 0, 0.1, 10, and 15 µM are used. Data was collected after ten days of treatment with salt and melatonin The performance of alfalfa seedlings with different concentrations of melatonin (0, 0.1, 10 and 15 µM) and salt stress (150 and 200 mM) treatment after ten days Effect of melatonin and NaCl on soluble carbohydrate (A), proline (B) and glycine betaine (C) of alfalfa root. Data are means of five replicates ± SD. Different letters indicate significant differences (P < 0.05) based on Duncan’s multiple range test. Melatonin concentrations of 0, 0.1, 10, and 15 µM are used. Data was collected after ten days of treatment with salt and melatonin Histochemical staining assay of ROS accumulations using DAB (A) and NBT (B) for investigated the effect of 0, 0.1, 10 and 15 µM melatonin and 0, 150 and 200 mM NaCl on H2O2 and O2− content in alfalfa root after ten days of treatment with salt and melatonin. Scale bar = 0.5 mm Effect of melatonin and NaCl on K+ (A), Na+ (B) and K+/Na+ content (C) of alfalfa root. Data are means of five replicates ± SD. Different letters indicate significant differences (P < 0.05) based on Duncan’s multiple range test. Melatonin concentrations of 0, 0.1, 10, and 15 µM are used. Data was collected after ten days of treatment with salt and melatonin Effect of melatonin and NaCl treatment on leaf melatonin content (A), leaf IAA content (B), root melatonin content (C), root IAA content (D), leaf IAA/melatonin ratio (E) and root IAA/melatonin ratio (F) of alfalfa. Data are means of five replications ± SD. Different letters indicate significant differences (P < 0.05) based on Duncan’s multiple range test. Melatonin concentrations of 0, 0.1, 10, and 15 µM are used. Data was collected after ten days of treatment with salt and melatonin Result of biplot principal components 1 and 2 analyses obtained from phytohormones and physiological parameters in alfalfa root subjected to melatonin and salinity (A), and the main objectives (B). © 2023, The Author(s), under exclusive licence to Plant Science and Biodiversity Centre, Slovak Academy of Sciences (SAS), Institute of Zoology, Slovak Academy of Sciences (SAS), Institute of Molecular Biology, Slovak Academy of Sciences (SAS).
Esfandiarpour, G.,
Mokhtari, M.,
Javadirad, S.M.,
Kolahdouzan, M.,
Almuslimawi, A. Scientific Reports (20452322)12(1)
Routine tissue-specific reference genes are often used in expression studies, but target genes are not taken into account. Using the relative RT-qPCR approach, we evaluated the expression of three target genes. At the same time, meta-analyses were conducted in various ethnic groups, genders, and thyroid cancer subtypes. When eight common reference genes were examined, it was discovered that some of them not only lacked consistent expression but also had considerable expression variance. It is worth noting that while choosing a reference gene, the mean gene expression and its standard deviation should be carefully addressed. An equation was developed based on this, and it was used to perform statistical analysis on over 25,000 genes. According to the subtype of thyroid cancer and, of course, the target genes in this investigation, appropriate reference genes were proposed. The intuitive choice of GAPDH as a common reference gene caused a major shift in the quantitative expression data of target genes, inverting the relative expression values. As a result, choosing the appropriate reference gene(s) for quantification of transcription data, and especially for relative studies of the expression of target gene(s), is critical and should be carefully considered during the study design. © 2022, The Author(s).
International Journal Of Reproductive Biomedicine (24764108)20(5)pp. 399-404
Background: Some previous human and animal studies have supported the idea that KDM3A down-regulation might be the main cause of male infertility, especially in non-obstructive azoospermia (NOA). The regulatory role of micro-RNAs (miRNA) has been investigated in the development of male infertility. Objective: The expression level of hsa-miR-30a-5p in azoospermia was evaluated to reveal its possible association with the etiology of male infertility. Materials and Methods: In this case-control study, 30 men with azoospermia (19 of whom had NOA) were selected as the case individuals, and 11 men with obstructive azoospermia (OA) were selected as control individuals. The best miRNA with the strongest ability to target the KDM3A gene was detected via comprehensive bioinformatics analysis. Reverse transcriptase quantitative polymerase chain reaction was used to assess the expression level of hsa-miR-30a-5p. After analyzing the data, the expression level of hsa-miR-30a-5p was compared between men with NOA and men with OA. Results: The findings supported the idea that hsa-miR-30a-5p is the miRNA with the best ability to target the KDM3A transcript. The expression analysis of hsa-miR-30a-5p indicated a significant overexpression (p = 0.04) in men with NOA compared to in men with OA. Conclusion: Hsa-miR-30a-5p was overexpressed in men with NOA compared to in control individuals. Hsa-miR-30a-5p could target the KDM3A transcript and may suppress its expression. © 2022, Research and Clinical Center for Infertitlity. All rights reserved.
Botanical Studies (1817406X)63(1)
Background: Alfalfa (Medicago sativa L.) is the most cultivated forage plant as a model in legumes. Salinity stress due to Na+ toxicity causes severe, oxidative stress as a main reason for program cell death (PCD) in plants. Melatonin application can increase plant productivity in response to diverse stressors via modulating plant antioxidant mechanisms and PCD inhibition in plants. Results: Alfalfa roots were subjected to different concentrations of in vitro salinity supplemented with melatonin (0.1, 10 and 15 µM) for ten days. Application of melatonin under salinity stress reduced ROS, H2O2 and O2- content and showed a dramatic impact on TTC reduction and augmented cell viability. Interestingly, melatonin inhibited caspase 3-like protease activity and could decrease DNA fragmentation induced by salinity while increased expression of anti-apoptotic genes BI-1, UCP1-UCP2 involved in PCD pathway. In contrast, in 300 mM salinity, γVPE gene as a proapoptotic of PCD down-regulated significantly. Conclusions: For the first time, present data showed that, melatonin plays a major function in preventing PCD in alfalfa root meristem cells. We attempted to offer a mechanism for the function of melatonin as an anti-apoptotic agent by demonstrating significant actions of melatonin on mitochondria proteins, such as UCPs, in a manner similar to animal cells. © 2022, The Author(s).
Biochemical Genetics (00062928)59(2)pp. 506-515
BCL2 apoptosis regulator (BCL2) is a cause of tumorigenesis whose CA-repeat promoter polymorphisms has inconsistent association with various types of cancers. The association of BCL2 polymorphism with breast cancer was investigated in the Isfahan province of Iran. PCRamplification of the CA-repeat was followed by polyacrylamide gel electrophoresis and direct sequencing for 120 breast cancer women and an equal number of corresponding healthy control individuals. Seven different alleles, ranging from 11 to 17 CA-repeats were observed. Short alleles with 11 to 14 repeats were protective (OR 0.363, P = 0.001), but large alleles with 15 to 17 repeats were threatening against breast cancer development (OR 2.780, P = 0.001). Accordingly, genotypes with large alleles showed a higher risk of breast cancer development (OR 3.400, P = 0.004). ERS1\ERBB2 positive breast cancer patients, but not PGRpositive ones, showed protection against breast cancer (OR 0.405, OR 0.346 respectively). In conclusion, women with at least one large allele of BCL2 were 3.4 times at higher risk of breast cancer development in the Isfahan province of Iran. © 2020, Springer Science+Business Media, LLC, part of Springer Nature.
Journal of Gorgan University of Medical Sciences (15624765)23(4)pp. 33-39
Background and Objective: Thyroid cancer is a common cancer of the endocrine system, and knowing the etiology can be effective in its treating. On the other hand, the FNA technique is not accurate enough, so finding a biomarker for thyroid cancer is of importance. This study was done to evaluate the NKX2-1 gene as indicator of differentiation of papillary thyroid carcinoma (PTC). Methods: In this case-control study, 17 fresh PTC tissue samples and 20 adjacent-healthy tissues were collected during thyroidectomy in Isfahan, Iran. RNA extraction was followed by cDNA synthesis. The expression of the NKX2-1 gene was performed using specific primers (exon-junction and intron spanning) using the RT-qPCR method. Results: An examination of the quality and quantity of extracted RNAs showed that they were intact and suitable for making cDNA. Examination of the melting curve showed a specific amplification of the NKX2-1 gene. The difference in expression of the NKX2-1 gene between PTC and healthy-adjacent tissues was 0.947. Conclusion: No difference in the expression of the NKX2-1 gene between the healthy tissue adjacent to the tumor and the tissue of the PTC tumor indicates that the PTC tumors were differentiated. © The author(s).
Scientific Reports (20452322)11(1)
The association of PRM1/2 with male azoospermia is well-documented, but the relationship between TXNDC2 deficiency and the azoospermia phenotype, sperm retrieval, and pathology has not been elucidated. Here we identified the association of TXNDC2 and protamines in evaluating testis pathology and sperm retrieval. An extensive microarray meta-analysis of men with idiopathic azoospermia was performed, and after undergoing several steps of data quality controls, the data passing QC were pooled and batch effect corrected. As redox imbalance has been shown to have a variable relationship with fertility, our relative expression studies began with candidate protamination and thioredoxin genes. We constructed a logistic regression model of TXNDC2 with PRM1 and PRM2 genes, and collective ROC analysis indicated a sensitivity of 96.8% and specificity of 95.5% with a ROC value of 0.995 (SE = 0.0070, 95% CI 0.982–1.000). These results demonstrate that TXNDC2, PRM1, and PRM2 combined have a robust power to predict sperm retrieval and correlate with severe azoospermia pathology. © 2021, The Author(s).
International Journal Of Reproductive Biomedicine (24764108)18(11)pp. 961-968
Background: The role of KDM3A and its downstream genes in male fertility has been approved in animal models. Additionally, the expression shrinkage of KDM3A is significantly correlated with human azoospermia phenotype. Aberrant expression of micro-RNAs could mislead spermatogenesis and mostly lead to diverse phenotypes of male infertility. Objective: The aim of this study was to evaluate the expression level of hsa-miR-27a-3p in azoospermic men to reveal its possible association with infertility. Materials and Methods: This case-control study was conducted on 30 azoospermic men, of whom, 19 had non obstructive azoospermia (NOA) and 11 obstructive azoospermia (OA) according to the pathological examinations. Comprehensive bioinformatics investigations were performed securely and hsa-miR-27a-3p was selected afterward. Reverse Transcriptase-quantitative polymerase chain reaction (RT-qPCR) method was used and statistical analysis was performed to compare the expression level of hsa-miR-27a-3p in both OA and NOA individuals. Results: In silico analysis suggested hsa-miR-27a-3p, with its potential binding ability to target KDM3A transcripts. The expression analysis of candidate hsa-miR-27a-3p indicated its significant overexpression in NOA men. Conclusion: The hsa-miR-27a-3p was overexpressed in NOA men compared to OA-control individuals. As a consequence, the overexpressed micro-RNA could downregulate directly KDM3A and indirectly TNP1 and PRM1. Therefore, spermatogenesis could be misled and male infertility could be developed. © Norioun et al.
Journal Of Kerman University Of Medical Sciences (20082843)27(5)pp. 426-435
Background: Hypertension with its related disorders is one of the most common health problems among the Iranian population. Hypertension can be developed by chronic stress and a positive association between stress and rs2596542 has been confirmed. Methods: A Total number of 112 hypertensive patients and 97 healthy individuals were involved in the study. Total blood genomic-DNA was extracted and PASA (PCR amplification of specific alleles) method was used to amplify MICA-rs2596542 polymorphic site. Different genotypes were visualized. The normality of the data was assessed and the binary logistic regression was used for OR and 95%CI calculations. Results: A-risk allele of rs2596542 increased the risk of hypertension development significantly (OR=1.734, p=0.006). Females were significantly more potent to hypertension development than males (OR=2.015, p=0.013). Risk-allele homozygotes (AA) showed a higher risk of hypertension development than GG (OR=2.132, p=0.020) and AG individuals (OR=3.206, p=0.006). Age adjustments at 70 years old, further increased the risk of hypertension development in GG (OR=3.772, p=0.011) and AG (OR=6.531, p=0.009) individuals. Conclusion: A-risk allele of rs2596542 could increase the risk of hypertension up to 3.2 folds and this risk could be upraised after sex and age adjustments. © 2020, Kerman University of Medical Sciences. All rights reserved.
Scientific Reports (20452322)10(1)
In RT-qPCR, accuracy requires multiple levels of standardization, but results could be obfuscated by human errors and technical limitations. Data normalization against suitable reference genes is critical, yet their observed expression can be confounded by pseudogenes. Eight reference genes were selected based on literature review and analysis of papillary thyroid carcinoma (PTC) microarray data. RNA extraction and cDNA synthesis were followed by RT-qPCR amplification in triplicate with exon-junction or intron-spanning primers. Several statistical analyses were applied using Microsoft Excel, NormFinder, and BestKeeper. In normal tissues, the least correlation of variation (CqCV%) and the lowest maximum fold change (MFC) were respectively recorded for PYCR1 and SYMPK. In PTC tissues, SYMPK had the lowest CqCV% (5.16%) and MFC (1.17). According to NormFinder, the best reference combination was SYMPK and ACTB (stability value = 0.209). BestKeeper suggested SYMPK as the best reference in both normal (r = 0.969) and PTC tissues (r = 0.958). SYMPK is suggested as the best reference gene for overcoming the pseudogene problem in RT-qPCR data normalization, with a stability value of 0.319. © 2020, The Author(s).
Iranian Biomedical Journal (2008823X)23(3)pp. 220-227
Background: KDM3A is a key epigenetic regulator expressed in the testis and is required for packaging and condensation of sperm chromatin. To this point, the association of the KDM3A gene with infertility has not been studied in human. The aim of this study was to screen any new mutation in KDM3A gene to explore more details of human male infertility. Methods: In this work, 150 infertile men (oligozoospermia and azoospermia) and 150 normal healthy fathers were studied. Polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and sequencing were used to screen any mutation in exons 12, 22, and 24 of KDM3A. Results: The infertile men showed various SSCP patterns for the exons 12 and 24, but not for exon 22. A transversion point mutation in exon 12 and a single nucleotide deletion in exon 24 were detected using sequencing analysis. The transversion mutation was located in the preceding exon of lysine-specific demethylase1 and Jumonji (Jmj)-C domain and the later one (deletion) in the cupin-like motif of KDM3A protein. Neither Y chromosome microdeletions nor partial azoospermia factor deletion was found in these patients. Conclusion: The mutations found in infertile men with otherwise unexplained severe spermatogenic failure could be considered as the origin of their abnormalities. © 2019, Pasteur Institute of Iran. All rights reserved.
Assad samani, L.,
Javadirad, S.M.,
Parsafar, S.,
Tabatabaeian, H.,
Ghaedi, K.,
Azadeh, M. Indian Journal of Clinical Biochemistry (09740422)34(4)pp. 485-489
Breast cancer as the second most common cancer worldwide tend to be experienced by Iranian women 10 years earlier with a peak incidence at the premenopausal stage. Genetic mutations of TP53 tumor suppressor gene has been shown to be related to early onset of breast cancer. It has been shown already that rs1625895 polymorphic site is related to glioma as well as lung cancer. In this study, we have investigated the role of rs1625895 polymorphism in breast cancer incidence in Iranian women. DNA extraction of 86 breast cancer patients and 96 control individuals have been used for allele-specific primer-PCR and genotyping of allele A and allele G of the TP53 rs1625895. Genotypes frequencies have been shown that GG homozygosis as the most frequent genotype is a significant association with increased risk of breast cancer development in Iranian women (odds ratio = 6, p = 0.002). On the other hand and in comparison to allele G, allele A could cause early death of breast cancer patients by threefolds significantly (p = 0.011). As a conclusion, we show that allele A is the minor allele in both breast cancer patients and also control individuals and major allele G, is related to the increased risk of breast cancer development in Iranian women. © 2018, Association of Clinical Biochemists of India.
Journal Of Kerman University Of Medical Sciences (20082843)25(3)pp. 191-197
Background: Prostate cancer is increasing among Iranian men and gene polymorphisms may play a role in the development of prostate cancer. Insulin-like growth factor 1 (IGF1) gene polymorphisms have been deeply explored in different malignancies. In this study, we aimed to explore the association of IGF1 CA repeat length polymorphism with the risk of prostate cancer development in Isfahan province of Iran. Method: The total blood of 100 prostate cancer patients and the equivalent matched control individuals were collected. DNA extraction was followed by IGF1 promoter polymorphism amplification. Genotyping was performed using polyacrylamide gel electrophoresis and sequencing was performed. Results: According to the results, IGF1 promoter polymorphic site showed six different alleles ranging from 17-22 CA repeats among our studied population. Comparing SL heterozygotes with both homozygotes, a significant increase in RR value (RR=4.5, p=0.031) was observed. Although age adjustment and family history did not elevate the RR value, but a significantly elevated risk of prostate cancer (RR= 3.143, p=0.002) was shown when we compared SS patients with LL ones according to their BPH history. Conclusion: In conclusion, carriers of (CA)17 allele could be at a higher risk of prostate cancer development and being SL heterozygotes could increase the risk of BPH development in our studied population. © 2018, Kerman University of Medical Scienc. All rights reserved.
Teimuri, S.,
Hosseini, A.,
Ghaedi, K.,
Tanhaei, S.,
Javadirad, S.M.,
Etemadifar, M.,
Nasr-esfahani, M.H. Gene Reports (24520144)12pp. 105-108
Multiple sclerosis (MS) is a multifactorial, chronic and inflammatory disease due to the myelin sheath damage of the central nervous system (CNS). Genome-wide association studies reveal that progression of MS is associated with single nucleotide polymorphisms (SNP). To elucidate the frequency of two SNPs (rs3745453 and rs1044165) with the incidence of MS, a population composed of 310 MS patients and 280 healthy volunteers were selected in Isfahan as a high prevalent location in the heart of Iran. After genomic extraction of blood samples, Allele specific primer PCR (ASP-PCR) was implemented for genotyping of the samples. Data indicated that frequency of carriage of CC genotype in rs3745453 in MS patients compared to healthy individuals and frequency of carriage of rs1044165 TT genotype increased in female MS patients compared to the controls. Logistic regression analyses indicated a risk factor role of rs1044165 and rs3745453 in the pathogenesis of MS. © 2018
Meamar, R.,
Javadirad, S.M.,
Chitsaz, N.,
Asadian ghahfarokhi, M.,
Kazemi, M.,
Ostadsharif, M. The Egyptian Journal Of Medical Human Genetics (20902441)18(3)pp. 225-230
Background Vitamin D plays an important role in neurodegenerative disorders as a crucial neuro-immunomodulator. Accumulating data provide evidences that vitamin D receptor (VDR) gene is a candidate gene for susceptibility to Parkinson's disease (PD). Aim To find out whether the risk of the development of sporadic PD might be influenced by VDR gene polymorphisms in an Iranian population or not. Subjects and methods A genetic study was conducted to investigate the relationship between VDR gene polymorphisms and the severity of PD. Fifty-nine PD patients and 53 matched-healthy controls were genotyped using polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) analysis. For this purpose, four single nucleotide polymorphisms (SNPs) in VDR gene including FokI T > C (rs 10735810), BsmI A > G (rs 1544410), ApaI A > C (rs 7975232), and TaqI C > T (rs 731236) have been evaluated. Results Our genotyping studies revealed that holding ApaI a allele and FokI f allele could significantly increase the risk of developing Parkinson's disease 1.85 and 2.46 times, respectively (p = 0.023 and 0.008). Moreover, Aa heterozygous of ApaI also shows a significantly elevated risk of developing PD when compared to AA homozygous (OR = 7.44, p = 0.005). For BsmI and TaqI polymorphisms, no significant difference in genotype or allele distribution was found between PD patients and the controls. Moreover, in this study, no significant association was found between different genotypes and Hoehn & Yahr staging and Unified Parkinson Disease Rating Stage (UPDRS) rating scale. Conclusion This study demonstrates a possible association between the VDR FokI and ApaI polymorphism and PD, indicating that VDR polymorphisms may change genetic susceptibility to sporadic PD in the Iranian population. © 2016 Ain Shams University
Journal of Isfahan Medical School (10277595)33(367)pp. 2407-2416
Background: Real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR), is a fast, sensitive and reliable method of gene expression comparison that is prone to a lot of technical errors. On the other hand, historical reference (housekeeping) genes are not suitable for all tissues. Herein, we have tried to identify and evaluate the best reference gene for testis tissues for further qRT-PCR experiments. Methods: Testis tissues of 15 men with non-obstructive (NOA) and 15 men with obstructive (OA) azoospermia (as control individuals) were collected. Primer designing and verification of four candidate reference genes including glyceraldehyde 3-phosphate dehydrogenase (GAPDH), ribosomal protein L37 (RPL37), ring finger protein 1 (RING1) and eukaryotic translation elongation factor 2 (eEF2) were performed using Beacon designer 8.1 software. PCR pre-optimization for reverse transcriptase input RNA and best primer concentration were included. Melt curve analysis was drawn and values of quantitation cycle (Cq) were extracted. Mean Cq analysis was calculated using BestKeeper v1 software and suitable reference genes were selected afterward. Findings: Comparing the mean Cq values between the NOA and OA groups declared that RPL37 and GAPDH showed the lowest standard deviations of 1.39 and 1.67 among the other candidates. GAPDH and RPL37 were selected as the best reference genes in testis tissues with their r values of 0.959 and 0.927, respectively. Conclusion: The results of this study show that the best reference genes for normalization of qRT-PCR data of testis tissues are GAPDH and RPL37. © 2016, Isfahan University of Medical Sciences(IUMS). All rights reserved.
Andrology (20472927)4(3)pp. 492-499
Summary: To evaluate the predictive value of histone demethylase KDM3A to protamine 1 (PRM1) mRNA expression ratio as a reliable marker of sperm retrieval in men with obstructive and non-obstructive azoospermia (NOA). Fifty eight azoospermic men, including 44 with NOA and 14 with obstructive azoospermia (OA). Testis tissue samples were collected from azoospermic men who have been referred for testicular sperm extraction (TESE) and micro-TESE. Relative expression ratio of KDM3A to PRM1 was analyzed after selection of approved reference genes. Histological classification of testis biopsies was performed. Sperm retrieval following TESE and micro-TESE was evaluated. A sperm retrieval prediction sensitivity of 95% was established when the Cq of PRM1 became smaller than the Cq of both KDM3A and GAPDH genes. However, azoospermic men with down-regulated KDM3A and decreased expression of PRM1 mRNA showed very low success for sperm retrieval (<25%), even after micro-TESE surgery. The KDM3A to PRM1 mRNA expression ratio can be used as a reliable marker of successful testicular sperm extraction in men with obstructive and non-obstructive azoospermia with 95% sensitivity. © 2016 American Society of Andrology and European Academy of Andrology.
Komeijani, M.,
Kardi, M.T.,
Javadirad, S.M.,
Naghshineh, N.,
Rezaei, M.,
Hemmati, S.,
Sichani, A.H. Journal of Isfahan Medical School (10277595)30(188)
Background: Human cytomegalovirus (HCMV) is a member of herpes virus family and its early diagnosis could be very critical in kidney transplant patients. The clinical manifestations of HCMV infection include a wide range from tissue rejection to death. The aim of this study was to compare the accuracy of two common methods of HCMV monitoring, i.e. real-time polymerase chain reaction (PCR) and pp65 antigen assay. Methods: We extracted the genomic DNA of 135 kidney transplant patients from their whole blood. HCMV monitoring was performed using TaqMan scanning probes and pp65 surface antigen detection by real-time PCR and pp65 antigen assay, respectively. Findings: According to our results, real-time PCR and pp65 antigen assay showed positive results in 36 (27%) and 29 patients (21%), respectively. In addition, the maximum and minimum rates of infection with HCMV were detected in spring (28%) and winter (16%), respectively. However, no significant association was established between HCMV infection and seasonal patterns. Conclusion: Our results indicated that real-time PCR was more accurate than pp65 antigen assay to detect HCMV infection in kidney transplant patients. We could also show that the maximum rate of infections with HCMV belonged to spring. However, the rates in the 4 seasons were not significantly different.
Journal of Isfahan Medical School (10277595)29(162)pp. 1937-1944
Background: Aromatase is a key enzyme in the synthesis of estrogen. This enzyme is encoded by the cytochrome P-450 (CYP19) gene. Overexposure of cells to estrogen causes breast carcinoma and breast cancer patients show a higher level of aromatase expression. The purpose of this study was to investigate polymorphism of the CYP19 intron 4 (TTTA)n among breast cancer patients and healthy individuals, and its correlation to risk of breast cancer. Methods: This study was a case-control study on 420 patients and 432 control women. DNA was extracted from the blood of the study subjects, and then the sequence was amplified by the polymerase chain reaction (PCR) technique. Thereafter, the number and sequence of TTTA was measured by electrophoresis in Polyacrylamide gel. Findings: The results of this study show that IGF-I gene allele distribution in the population of Isfahan varies between 7-13 repetitions. The most common allele in both controls and cases was (TTTA)7-TCT with frequencies of 34.3% and 34.8%, respectively. Conclusion: The results of the current study show that the frequency distribution of 10 TTTA repeats is higher in the controls (2.3%) than in patients (1.4%). In other words, not only does 10 TTA not in-crease the rate of breast cancer, but it may even cause people with this allele to be more resistant to cancer. © 2012, Isfahan University of Medical Sciences(IUMS). All rights reserved.
