The study of association between reduced folate carrier 1 (RFC1) polymorphism and non-syndromic cleft lip/palate in Iranian population

Introduction: Cleft lip/palate is one of the most common congenital defects and is supposed to have multifactorial etiology, including a complex interaction between genetics and environment. Reduced folate carrier 1 (RFC1) gene takes part in folate transportation within the cells. In this study, the association of A80G polymorphism in the RFC1 gene with the non-syndromic cleft lip/palate (nsCL/P) was investigated in Iranian infants for the first time. Methods: In this case-control survey, 122 Iranian infants with nsCL/P and 164 healthy infants were investigated for RFC1 polymorphism by PCR and RFLP methods. The results were statistically compared with control group, odds ratios with 95% CI were estimated by univariate and multivariate logistic regression model and a P <0.05 was considered statistically significant. Results: The RFC1 G allele was significantly higher (P=0.001; OR=7, 95% CI: 4.7-10.2) in the cases (60.3%) compared with the controls (17.9%). Not only the RFC1 AG genotype was significantly higher (P<0.001; OR=44, 95% CI: 14.6-133) in cases (67.8%) than the controls (27.4%), but also GG genotype (P<0.001; OR=85, 95% CI: 20.5-352) was much higher in cases (26.4%) than the controls (4.3%). Conclusion: Our study indicated that the RFC1 (A80G) polymorphism was associated with the nsCL/P in Iranian population. Moreover, 80GG homozygosity was significant in the cases. The presence of G allele can be considered as a risk factor for the nsCL/P. Infants with the GG and AG genotypes were more prone to cleft lip/palate as compared to the AA ones. This finding emphasizes the role of RFC1 gene and the intracellular levels of folate

Comparing four laboratory three-parent techniques to construct human aged non-surrounded nucleolus germinal vesicle oocytes: A case-control study

Background: The three-parent assisted reproductive technique may increase oocyte competence. Objective: In this case-control study, the suitability of germinal vesicle transfer (GVT), synchronous ooplasmic transfer (sOT), asynchronous ooplasmic transfer using cryopreserved MII oocyte (caOT), and asynchronous ooplasmic transfer using waste MII oocyte (waOT) for maturation of the human-aged non-surrounded nucleolus germinal vesicle-stage (NSN-GV) oocyte were investigated. Materials and Methods: NSN-GV oocytes were subjected to four methods: group A (GVT), B (sOT), C (caOT) D (waOT), and E (Control). The fusion rates, MI, MII, ICSI observations and cleavage at 2-cell, 4-cell, and 8-cell stages were compared in the groups. Results: In GVT, none of the oocytes fused. In sOT, all oocytes fused, 20 achieved the MI, 14 progressed to MII, 8 fertilized, 6 cleaved and 5, 4, and 3 achieved the 2- cells, 4-cells and 8-cells, respectively. In caOT, all oocytes fused and achieved the MI, 8 progressed to MII and fertilized, 6 cleaved and 6, 5, and 5 achieved the 2-cells, 4- cells, and 8-cells respectively. In waOT, all oocytes fused, 5 and 3 progressed to MI and MII, respectively, but only one fertilized, cleaved and reached a 4-cells stage. In group E, 6 and 2 oocytes progressed to MI and MII, respectively, and only one fertilized but arrested at the zygote stage. caOT had the highest survival rate when compared to sOT (p = 0.04), waOT (p = 0.002), and control (p = 0.001). Conclusion: The caOT method was beneficial over sOT, waOT, and GVT in supplementing the developmental capacity of human-aged NSN-GV oocytes. Key words: Assisted reproductive techniques, In vitro oocyte maturation techniques, Nuclear transfer techniques, Oocytes, Oocyte donation

Analysis of the genes encoding the spp24 protein in human and mouse and identification of interacting proteins

Secreted phosphoprotein 24 (spp24) is a member of the cystatin superfamily.;This study identifies a rare single-amino acid polymorphism (p.S38F) of human spp24 and its importance has been assessed by comparing the sequence of human spp24 with that of eight other species.;The gene encoding spp24 in mouse (Spp2), like its human ortholog, comprises eight exons with an apparently TATA-les promoter. The exon-intron structure is identical in mouse and human and the size and location of intron 1 is conserved between many species. Using several strategies, the gene encoding spp24 in mouse has been mapped to 88832387-88853226 bp of the mouse chromosome 1.;An extensive expression study was carried out on the mouse and human genes encoding spp24. These studies indicated that the gene has an expression pattern of a tissue-specific and cell-specific nature, being expressed predominantly in liver and its expression is down-regulated by lipopolysaccharide (LPS) and tumour necrosis factor alpha (TNFalpha).;In an attempt to elucidate the function of the spp24 protein in mouse, a pooled-tissues cDNA library was constructed in a yeast two-hybrid vector. Two different constructs comprising the entire spp24 protein and the C-terminal non-cystatin like domain of the protein were used individually as baits in the yeast two-hybrid system to screen the constructed library. Seven potential interacting proteins were identified including granulin precursor also known as acrogranulin/epithelin (Grn), tissue specific transplantation antigen P35B (Tsta3), keratin complex 1, acidic, gene 18 (Krt1-18), keratin complex 1, acidic, gene 13 (Krtl-13 ), vimentin (Vim), similar to protein phosphatase 1, regulatory (inhibitor) subunit 12C (no gene symbol yet assigned) or myosin binding subunit 85 and alpha-actinin-4 (Actn4)

Study on Association Between GSTP1 (rs1695) and Late-Onset Alzheimer Disease and Interaction With APOe4

Objectives GSTs are detoxification enzymes that remove excess reactive oxygen species (ROS) from cells. Evidence suggests that oxidative stress plays a role in several stages of the neurodegenarative disease like Alzheimer disease. Free radicals and similar molecules are classified as reactive oxygen species (ROS), which can cause oxidative modifications in the cell. In this study we have investigated the association between GSTP1 (rs1695) and AD risk for  genetic variant in Iranian population. Methods & Materials The patient group consisted of 280 cases for GSTP1 gene investigation, whose Alzheimer disease had been approved by psychologists based on clinical test (DSM-IV). The control group included 168 healthy individuals, satisfying the condition of not having any psychological disorders. Individuals’ genotype have been determined by PCR-RFLP method. Statistical analysis was done by logistic regression using OpenEpi 2.3.1 and SPSS 16. Results Significant association was observed between heterozygote genotype (AG) rs1695 A/G of GSTP1 gene and the risk of Alzheimer disease (P=0.005, OR=0.57[0.38-0.84]).This genotype acts as a protective factor. This observed result was significant in within women group (P=0.02). Significant interaction was also found between heterozygote genotype (AG) rs1695 A/G of GSTP1 gene (protective factor) and absent ε4 allele in our study group (P=0.001). Conclusion Based on our results, we suggest that heterozygote genotype (AG) rs1695 A/G of GSTP1 gene can act as a protective factor for Alzheimer disease in Iranian population.&nbsp

Investigation of DRD2 and HTR2A mRNA Expression in Two Therapeutic States of Antipsychotic Polypharmacy and Aripiprazole Monotherapy in the Peripheral Blood of Patients With Schizophrenia

Objectives: Schizophrenia is a severe psychiatric disorder that has profound effects on both individuals and the community. Notwithstanding the suggestion for treating schizophrenia with a minimum dose of drugs, antipsychotic polypharmacy increases the patient’s care costs and drug interactions. Aripiprazole reduces the metabolic side effects of antipsychotic polypharmacy treatment. DRD2 and HTR2A can serve as predictors for response to treatment in schizophrenic patients. The purpose of this survey was to measure the DRD2 and HTR2A genes expression in the peripheral blood samples using Real-Time Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Methods: A total of 19 patients with a long history of schizophrenia who received at least two types of antipsychotics with daily doses of more than 500 mg of chlorpromazine were entered into the study. The response rates to the treatment based on scores in the Brief Psychiatric Rating Scale (BPRS) questionnaire and DRD2 and HTR2A expression were compared between antipsychotic polypharmacy status and 6 months after monotherapy with aripiprazole. Results: The levels of DRD2 expression decreased significantly after the intervention. The mean changes in HTR2A expression and the BPRS questionnaire and also the relationship between changes in DRD2 and HTR2A expression and changes in BPRS score after the intervention were not significant. Discussion: The conversion of the antipsychotic polypharmacy state to monotherapy with aripiprazole has been accompanied by a significant decrease in DRD2A expression. These genes can be used for evaluating the response rate of schizophrenia treatments in the future

Identification of Genetic Polymorphism Interactions in Sporadic Alzheimer’s Disease Using Logic Regression

Objectives: Genetic polymorphism interactions are among the important factors in affliction with complex diseases like Alzheimer’s disease. The important goal of genetic association studies is to identify a combination of polymorphisms and measure their importance in increasing the risk of occurrence of such diseases. In this study, feature selection approach of logic regression was used to identify the interactions among genetic polymorphisms influential in patients affected with Alzheimer’s disease. Methods: 101 Alzheimer’s cases and 109 control subjects from Iranian population were recruited in a case-control study. The evaluation of genes in two groups was performed using molecular technique methods in particular, the PCR-RFLP technique was used to evaluate the intended polymorphisms in APOE, ABCA1, CALHM, CCR2, GSK3β, SAITOHIN, TAU, TNF-α and VDR genes, and then the feature selection approach was used to detect the significance polymorphisms and interactions between them. Results: Based on feature selection approach, the two-way interaction between the polymorphisms of SAITOHIN and APOE genes were significant on occurrence of Alzheimer’s disease. Discussion: Logic regression approach is recommended to detect interaction in the genetic association studies

Early Fetal Gender Determination Using Real-Time PCR Analysis of Cell-free Fetal DNA During 6th-10th Weeks of Gestation

Nowadays, new advances in the use of cell free fetal DNA (cffDNA) in maternal plasma of pregnant women has provided the possibility of applying cffDNA in prenatal diagnosis as a non-invasive method. In contrary to the risks of invasive methods that affect both mother and fetus, applying cffDNA is proven to be highly effective with lower risk. One of the applications of prenatal diagnosis is fetal gender determination, which is important in fetuses at risk of sex-linked genetic diseases. In such cases by obtaining the basic information of the gender, necessary time management can be taken in therapeutic to significantly reduce the necessity of applying the invasive methods. Therefore in this study, the probability of detecting sequences on the human Y-chromosome in pregnant women has been evaluated to identify the gender of fetuses. Peripheral blood samples were obtained from 80 pregnant women with gestational age between 6th to 10th weeks and the fetal DNA was extracted from the plasma. Identification of SRY, DYS14 & DAZ sequences, which are not presentin the maternal genome, was performed using Real-Time PCR. All the obtained results were compared with the actual gender of the newborns to calculate the test accuracy. Considerable 97.3% sensitivity and 97.3% specificity were obtained in fetal gender determination which is significant in the first trimester of pregnancy. Only in one case, false positive result was obtained. Using non-invasive method of cffDNAs in the shortest time possible, as well as avoiding invasive tests for early determination of fetal gender, provides the opportunity of deciding and employing early treatment for fetuses at risk of genetic disease