Association of DRD2, DRD4 and COMT genes variants and their gene-gene interactions with antipsychotic treatment response in patients with schizophrenia.

Antipsychotic drugs are the first line of treatment in schizophrenia; although antipsychotic responses indicate a wide interindividual variety in patients with schizophrenia. This study aimed to investigate the association between four polymorphisms in DRD2, DRD4 and COMT genes and their gene-gene interactions with antipsychotic treatment response in patients with schizophrenia. A total of 101 patients with schizophrenia were recruited and stratified in treatment responder and treatment resistant groups based on the published criteria of resistant to treatment using PANSS. Clinical and demographic factors were analyzed. Genomic DNA was extracted from whole blood and genotyping for the four polymorphisms were done by ARMS-PCR, PCR-RFLP and gap-PCR. Gene-gene interactions were analyzed by logistic regression. In case of DRD2 A-241G, G allele was significantly associated with resistant to treatment. Regarding DRD4 120-bp duplication, 240/240 genotype was significantly associated with resistant to treatment comparing to other genotypes in a dominant model. The genotype combination of DRD4 240/240 and COMT Val/Val was significantly associated with treatment resistant. Among DRD2 AA genotype, COMT met allele carriers which also had a 120 bp allele of DRD4 had a significantly better response to antipsychotics. Moreover, analysis of clinical and demographic factors demonstrated a significantly longer duration of hospitalization and higher chlorpromazine-equivalent daily dose in resistant to treatment patients. Discovering the polymorphisms which effect treatment response to antipsychotics will provide the possibility of genetic screening before starting an antipsychotic treatment which enhances the chance of responding to antipsychotics and decreases drugs side effects and costs.

Evaluation of BRCA1 Gene Promoter Methylation Status in Sporadic Breast Cancer Patients in Southwest of Iran.

OBJECTIVE: Similar to other types of cancer, the development of breast cancer is a multi-stage process, consisting of various mutations and epigenetic changes in many genes. Mutations in the BRCA1 gene, which is a tumor suppressor gene, are considered as the most important types of mutations. The pivotal role of epigenetics is currently considered as the primary key to carcinogenesis. Several studies have previously reported the BRCA1 epigenetic silencing through promoter methylation in the pathophysiology of breast cancer cells. This study aimed to investigate whether the BRCA1 gene promoter methylation in peripheral blood cells is correlated with the risk of breast cancer. METHODS: In the current study, DNA samples were extracted from blood cells belonged to 74 patients with breast cancer as well as 30 healthy individuals, and the BRCA1 gene promoter methylation status in these two groups was examined using Methylation Specific PCR (MSP). RESULT: out of 74 patients, 2 cases demonstrated methylation in their BRCA1 gene promoter; however, none of the healthy controls demonstrated methylation status. Among these 74 patients, 13 cases were at the early stages (stage I), and two patients who had BRCA1 gene methylation (15.4%), were in this group (p=0.02). While 34 and 27 patients were at stages II and III, respectively, showing a negative state of BRCA1 gene methylation. CONCLUSION: Although 2 out of 74 patients resulted positive for methylation status, the healthy controls demonstrated no methylation. Consequently, there was inadequate evidence to confirm the association between BRCA1 gene promoter methylation in blood and the risk of developing breast cancer.

Molecular diagnosis of maturity onset diabetes of the young in Iranian patients: improving management.

BACKGROUND: The purpose of this study is to identify the mutations of the most common form of maturity-onset diabetes of the young (MODY), also known as MODY3, in diabetic patients suspected of MODY. This can recommend appropriate medical surveillance of at-risk family members of MODY based on the genetic cause. METHODS: We analyzed the clinical course of 19 patients from 12 unrelated Iranian families with diabetes features. The coding regions and intron-exon boundaries of the hepatocyte nuclear factor 1 alpha (HNF1A) gene were studied by polymerase chain reaction (PCR) and sanger sequencing. Also, the detected mutation was analyzed by bioinformatics tools. RESULTS: One novel frameshift insertion mutation (p.Glu11Argfs*12) was detected in one of the probands and seven other patients of her family with the heterozygote state. The mutation is located in the exon1 of the dimerization domain of the HNF1A gene. According to the In Silico analysis, the detected mutation is predicted as a pathogenic one. CONCLUSIONS: Differential diagnosis between MODY3 and other forms of diabetes can be considered a necessity in terms of overlapping symptoms of MODY3 with type1 or 2 diabetes. Molecular genetic testing can provide an accurate diagnosis for optimal management.

DNA vaccine containing Flagellin A gene induces significant immune responses against Helicobacter pylori infection: An in vivo study.

OBJECTIVES: Helicobacter pylori is one of the most prevalent human infectious agents that is directly involved in various upper digestive tract diseases. Although antibiotics-based therapy and proton pump inhibitors eradicate the bacteria mostly, their effectiveness has been declined recently due to emergence of antibiotic-resistant strains. Development of a DNA vaccine is a promising approach against bacterial pathogens. Genes encoding motility factors are promising immunogens to develop a DNA vaccine against H. pylori infection due to critical role of these genes in bacterial attachment and colonization within the gastric lumen. The present study aimed to synthesize a DNA vaccine construct based on the Flagellin A gene (flaA), the predominant flagellin subunit in H. pylori flagella. MATERIALS AND METHODS: The coding sequence of flaA was amplified through PCR and sub-cloned in the pBudCE4.1 vector. The recombinant vector was introduced into the human dermal fibroblast cells, and its potency to express the flaA protein was analyzed using SDS-PAGE. The recombinant construct was intramuscularly (IM) injected into the mice, and the profiles of cytokines and immunoglobulins were measured using ELISA. RESULTS: It has been found that flaA was successfully expressed in cells. Recombinant-vector also increased the serum levels of evaluated cytokines and immunoglobulins in mice. CONCLUSION: These findings showed that the pBudCE4.1-flaA construct was able to activate the immune responses. This study is the first step towards synthesis of recombinant-construct based on the flaA gene. Immunization with such construct may inhibit the H. pylori-associated infection; however, further experiments are urgent.

Association between Endothelial nitric oxide synthase and Hepatic lipase gene polymorphisms with the risk of coronary artery disease in Southern Iran population - A case control study.

Coronary artery disease is a multifactorial genetic disease caused by the interaction between genetic and environmental factors. Angiography is the gold standard method for the diagnosis and determining the stage of cardiac disorder. The rs1800588 at the Hepatic Lipase gene and rs1799983 at the endothelial nitric oxide synthase (eNOS) gene are two candidate SNP that result in increased risk of this disease. The aim of this study was to find out the associations of the two mentioned polymorphisms with angiographically proven coronary artery patients in a southern Iranian population. In this study, this two polymorphisms in 287 patients and 229 matched controls were confirmed by angiography and analyzed. Genotype analysis was carried out by PCR and RFLP. Data showed that a significant difference for the eNOS gene polymorphism (p = 0.004) and a non-significant difference for the Hepatic lipase polymorphism (p = 0.261) and increasing severity of angiographic evidences of coronary artery disease were observed. Conclusively the significant association of the G894T with the narrowing of two or three coronary vessels of this patients in an Iranian population have been detected.

A DNA Repair Pathway Polymorphism (rs25487) and Angiographically Proven Coronary Artery Patients in a Population of Southern Iran.

BACKGROUND: Coronary Artery Disease (CAD), which is a multifactorial genetic disease, is known as one of the most common causes of death worldwide. In this regard, X-ray Repair Cross-Complementing group 1 (XRCC1), a DNA repair protein involved in Single-Strand Breaks (SSBs), and Base Excision Repair (BER) pathways have been reported to be responsible for the efficient repair of single strand breaks and damaged bases in DNA. OBJECTIVES: In the current study, we analyzed Arg399Gln (rs25487), which is one of the most common polymorphisms of XRCC1 gene that might be associated with the increased risk for CAD. METHODS: This case-control study was performed to investigate the relationship between this polymorphism and CAD development. In this study, 290 patients and 216 controls were diagnosed by cardiac angiography and then screened for the above-mentioned polymorphism using Restriction Fragment Length Polymorphisms (RFLP) method. RESULTS: The frequency of the GA genotype of XRCC1 Arg399Gln (rs25487) was significantly higher in CAD patients compared to the controls (p=0.002, OR: 1.21, 95% CI: 1.06-1.37). Moreover, its dominant mode (AA + GA) genotype had a 1.851-fold increase in the risk of CAD (p = 0.005). CONCLUSION: Our findings demonstrated that Arg399Gln polymorphism of XRCC1 (rs25487) has a significant relationship with CAD and also plays a probable predisposing role in that. Our results support the role of DNA damages and the malfunctions of DNA repair system in the patients with CAD.

Increased Expression Level of Long Noncoding RNA H19 in Plasma of Patients with Myocardial Infarction.

Long noncoding RNAs (lncRNAs) are lengthy noncoding transcripts which are actively involved in crucial cellular pathways. Tissue-specific expression of lncRNAs besides its secretion into the body fluids, has made lncRNAs in attention as biomarkers of the diseases. According to the role of lncRNAs, especially H19 in cardiac regeneration, it is not surprising if their altered expression levels lead to cardiac diseases. In the present study, the relative expression of H19 was compared in the plasma of atherosclerotic myocardial infarction and control individuals by real time-PCR, and data were normalized using GAPDH. The association of plasma level of lipid and homocystine with H19 expression was also considered. The potential of H19 to discriminate the case from control was studied using the ROC analysis. We found that the plasma level of H19 transcript significantly increased in the plasma of patients in comparison with the control group. Additionally, the relative expression level of H19 was directly associated with the plasma homocystine level. The relative expression of H19 at threshold of 0.3 showed 70% sensitivity and 94% specificity to discriminate cases from controls. This study revealed that the expression level of H19 may be considered as a biomarker of myocardial infarction, although further studies are needed to generalize this finding.

Influence of Two DNA Repair Pathway Polymorphisms in Colorectal Cancer Risk in Southwest Iran.

OBJECTIVE: X-ray cross-complementing group 1 (XRCC1) and 8 Oxo guanine DNA-glycosylase 1 (OGG1) genes are implicated in the repair of single-stranded breaks (SSBRs) and base excision repair (BER) pathways. Common polymorphisms in DNA repair genes are supposed to decrease the capability of DNA repair and cause genetic instability. This study was designed to investigate the association between XRCC1 (rs25487) and OGG1 (rs1052133) polymorphisms and susceptibility to colorectal cancer (CRC) in the Ahvaz city, south-west Iran. METHODS: This case- control study comprised 150 patients and 150 controls that were selected from 2 educational hospitals in Ahvaz. They were matched for age and gender, and their genotyping was carried out by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Our results indicate that the frequency of the Gln (A) allele of XRCC1 (rs25487) is significantly higher in colorectal cancer patients, compare to controls (p = 0.01, OR: 1.54, 95% CI 1.9-13.3). Significant increased risk of cancer was observed in XRCC1 (rs25487) genotypes (p = 0.001 OR: 5.3, 95% CI 1.9-14.2 for Gln / Gln), while no association was found between OGG1 (rs1052133) and colorectal cancer risk (p = 0.6). CONCLUSION: Our study suggests that XRCC1 (rs25487) polymorphism might be associated with an increasing risk of CRC in Ahvaz. It also demonstrates positive correlation between the XRCC1 (rs25487) genotypes and demographic characteristics, such as smoking and increased age in patients and control groups.
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Double Trisomy 48,XXY,+21 in a Neonate with Congenital Heart Disease.

Double trisomy 48, XXY, +21 or Down-Klinefelter syndrome is a rare occurrence and presents clinical manifestation of trisomy 21 in early life and of Klinefelter syndrome after 10 months of age. The phenotypic and karyotyping characteristics of a 2-month-old boy were reported. He had mild clinical feature of Down syndrome and echocardiographic features of atrioventricular (AV) septal defects with severe pulmonary valve stenosis.

Study of the immunogenicity of outer membrane protein A (ompA) gene from Acinetobacter baumannii as DNA vaccine candidate in vivo.

OBJECTIVES: Acinetobacter baumannii is one the most dangerous opportunistic pathogens in hospitalized infections. This bacterium is resistant to 90% of commercial antibiotics. Therefore, developing new strategies to cure A. baumannii-infections is urgent. The DNA vaccines new approach in which the immunogen can be directly expressed inside the target cells through cloning of immunogen into an expression vector. The outer membrane protein A(OmpA) is one the critical factors in pathogenicity of A. baumannii which has been repeatedly described as a powerful immunogen to trigger the immune responses. As the pure form of the OmpA is insoluble, vaccine delivery is very hard. MATERIALS AND METHODS: We previously cloned the ompA gene from A. baumannii into the eukaryotic expression vector pBudCE4.1 and observed that the OmpA protein has been considerably expressed in eukaryotic cell model. In current study, the immunogenic potential of pBudCE4.1-ompA has been evaluated in mice model of experimental. The serum levels of IgM, IgG, IL-2, IL-4, IL-12 and INF-γ were measured by enzyme-linked immunosorbent assay (ELISA) after immunization with ompA-vaccine. The protective efficiency of the designed-DNA vaccine was evaluated following intranasal administration of mice with toxic dose of A. baumannii. RESULTS: Obtained data showed the elevated levels of IgM, IgG, IL-2, IL-4, IL-12 and INF-γ in serum following the vaccine administration and mice who immunized with recombinant vector were survived more than control group. CONCLUSION: These findings indicate ompA-DNA vaccine is potent to trigger humoral and cellular immunity responses although further experiments are needed.

Association study of rs10768683 and rs968857 polymorphisms with transfusion-dependent thalassemia (TDT) in a southern Iranian population.

Previous studies reported that detection of polymorphisms inherited through paternal model could be potential markers for the Non-Invasive Prenatal Diagnosis (NIPD) of ß-thalassemia. The aim of the current study was to find out the associations of rs10768683 and rs968857 with transfusion-dependent thalassemia (TDT) in a southern Iranian population. A total of 175 subjects were investigated, divided into patients with TDT as case group (n = 75) and healthy people as control group (n = 100). Genomic DNAs were extracted from peripheral blood using salting out procedure. Genotyping rs10768683 and rs968857 was carried out by ARMS-PCR, then statistical analyses were assessed using SPSS, and Medcalc ver. 18 software. Data showed that rs10768683 was statistically significant in co-dominant model of inheritance (P = 0.025, OR = 2.11 [1.08-4.15]) and genotype frequencies of CG among controls and cases were 0.68 and 0.80, respectively. However, according to genotype frequencies, there was no association between rs968857 and TDT among cases and healthy controls in any models of inheritance. In conclusion, the present study showed the association of rs10768683 with major ß-thalassemia through ARMS-PCR technique.

Pattern Analysis of Short Tandem Repeats Allele Frequencies among the Population of Khuzestan Province, South of Iran.

BACKGROUND: The basis of genetic fingerprinting and DNA profiling in forensic laboratories is the use of Short Tandem Repeats (STRs) according to local and ethnical genetics characteristics. METHODS: Forensic parameters and allele frequencies for 15 autosomal STRs in 100 unrelated individuals from Khuzestan province, south Iran were determined. PCR was carried out for amplification of STRs and GeneMapper ID software was used for genotyping and allelic analyzing. RESULTS: The Power of Exclusion (PE) varied between 0.332 (TPOX) and 0.768 (FGA). With exception of the THO1 (0.020), TPOX (0.014) and D18S51 (0.003), other STRs showed no deviation from the Hardy-Weinberg equilibrium (p>0.05). CONCLUSION: Out of 15 STRs, 12 repeats seemed to be more useful and more powerful tools in identity and paternity determination for our studied population. Variation in our data analysis revealed that effective use of these 15 STR loci in forensic cases needed to be localized by collection and analysis of population data from the general population.

The effect of question generation activity on students' learning and perception.

INTRODUCTION: Student-generated questions can be a very helpful tool in medical education. The use of this activity can allow the students to feel more involved in the subjects covered and may improve their knowledge and learning. The aim of this study was to identify the effect of question-writing activity as a stimulus factor on learning in midwifery students and determine their perception about this activity. METHODS: This quasi-experimental study with two groups of pre- and post-tests was conducted on two groups of midwifery students who had taken the immunology course. Two classes of midwifery students (N=62) participated and were randomly assigned to two different groups. One class was selected as the experimental group (n=32) and the other class was considered as the control group (n=30). The experimental group's students were asked to write questions covering different topics of the syllabus components taught during 15 weeks from February 2016 to May 2016. They were asked to write, answer and explain their multiple-choice questions (MCQs). The students' performance in immunology course was compared between the two groups at the end of the semester. After their final exam, we asked them to fill in a questionnaire on their views about this activity. The data were analyzed by independent t- test using SPSS software, version 18. RESULTS: The differences between pre- and post-test mean scores of the experimental and control groups were 24.53±5.74 and 20.63±5.58, respectively. The results of independent t-test showed that these differences in the two groups were significant (p=0.009). Nevertheless, most of the students stated that question-writing activity as a learning tool is an unfamiliar exercise and unpopular learning strategy. CONCLUSION: Results showed that question writing by students has been found to promote learning when it is implemented as a part of the teaching curriculum in immunology course; therefore, this activity could be effective in improving the students' learning.

Cloning of ompA gene from Acinetobacter baumannii into the eukaryotic expression vector pBudCE4.1 as DNA vaccine.

Antibiotic resistant features of Acinetobacter baumannii is partly due to the decreased outer membrane proteins (OMPs) permeability. The OmpA is one of the most conserved proteins among A. baumannii with a considerable antigenic potential to stimulate the multidimensional immune system responses. The present study was aimed to clone the ompA gene into the eukaryotic expression vector with potential as DNA vaccine. The ompA gene of A. baumannii was amplified using polymerase chain reaction (PCR). The target DNA was cloned and sub-cloned into the pTZ57R/T and pBudCE4.1 vectors, respectively. The recombinant vectors containing ompA were then validated using colony PCR, vector sequencing and double-digestion strategies. The pBudCE4.1-ompA recombinant plasmid was transfected into the human dermal fibroblast cells (HDF) and presence of ompA transcript and protein was evaluated using reverse transcribed-PCR (RT-PCR) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Our finding from colony PCR, sequencing and enzyme double digestion result confirmed that target gene has been successfully inserted into the pTZ57RT and pBudCE4.1. The presence of an expected band (1112 bp) in RT-PCR as wells as a ~ 38 kDa band during SDS-PAGE showed that the recombinant pBudCE4.1-ompA construct was efficiently transfected into the HDF cells and expressed. Altogether, our observation demonstrated that the recombinant pBudCE4.1-ompA construct was successfully produced although further experiments are needed.

The recurrence risk of genetic complex diseases.

Complex inherited diseases affected by an interaction between collective effects of the genotype at one or multiple loci either to increase or to lower susceptibility to disease, combined with a variety of environmental exposures that may trigger, accelerate, exacerbate, or protect against the disease process. The new aspects of genetic techniques have been opened for diagnosis and analysis of inherited disorders. While appropriate Mendelian laws is applied to estimate the recurrence risk of single gene diseases, using empirical recurrence risks are the most important and available method to evaluate pedigree of complex (multifactorial), chromosomal, and unknown etiology disorders. Although, generally, empirical recurrent risks are not accurate, either because of the difference of gene frequencies and environmental factors among populations or heterogeneity of disease; using results of plenty family population studies, computerized estimating programs, genotyping technologies, and Genome-wide association studies (GWASs) of single nucleotide polymorphisms (SNPs), can make it possible nowadays to estimate these risks. The specific family situation and importance recurrence risks of some common complex genetic diseases will be presented in this review and some important multifactorial disorders' recurrence risks will be summarized to help genetic counselors for supporting families and representing better view of genetic disorders.

Medical genetics teaching in Iranian medical schools, especially Ahvaz, south of Iran.

INTRODUCTION: Physicians have to visit, diagnose and refer patients with genetic disorders, so they need to be familiar with the basics and indications of genetic tests. In other words, they should have effective theoretical and practical knowledge about medical genetics before they do their job. Medical genetics courses at Medical Universities of Iran are generally presented as a theoretical subject in the first period of medical education. METHODS: In this descriptive research, the results of interviews with teachers of medical genetics in 30 medical schools in Islamic Republic of Iran and responses to a questionnaire by 125 medical students of Ahvaz Jundishapour University of medical sciences, about presentation time, curricula and also efficacy of medical genetics courses were analyzed. The interviews with teachers were done on phone and the students' comments were collected by a researcher-made questionnaire. The data were analyzed, using SPSS software, version 14.  RESULTS: In two thirds of medical universities, medical genetics is taught in the third or fourth semester and in 5 universities in the fifth semester. 86% of the students believed that the quality of genetics courses is moderate and such courses are very beneficial to medical students. CONCLUSION: This article suggests that medical genetics be offered in the second or third period of medical education (physiopathology or stagger period). Furthermore, in teaching such courses advanced educational methods (animation presentation, case-based learning, problem-based learning, etc.) should be used, together with simple genetic tests in laboratories, and the visit of genetic patients in hospitals and genetics centers.

An understanding of the genetic basis of asthma.

Asthma is the most common chronic childhood disease in developed nations and its prevalence has increased in the world over the last 25 years. It is a complex disease with both genetic and environmental risk factors. Asthma is caused by multiple interacting genes, some having a protective effect and others contributing to the disease pathogenesis, with each gene having its own tendency to be influenced by the environment. This article reviews the current state of the genetics of asthma in six categories, viz. epidemiology, management, aetiology, family and twin studies, segregation and linkage studies, and candidate genes and single nucleotide polymorphisms (SNPs).

Association of IL-4 and ADAM33 gene polymorphisms with asthma in an Indian population.

There are more than 100 candidate genes of asthma located on 23 human chromosomes. Interleukin-4 (IL-4), located on chromosome 5q31, and ADAM33, located on chromosome 20p13, and some single nucleotide polymorphisms (SNPs) of these genes have been shown to be associated with asthma and its manifestations in different populations. The most prominent SNPs of IL-4 and ADAM33 are 589C>T and 400A>G, respectively. There are also controversial reports on the association of these SNPs with asthma. In the present study, we analyzed these two SNPs in 100 patients with asthma and 50 controls through PCR amplification and restriction digestion to evaluate association of these two SNPs with asthma. The nonsignificant differences were observed for the IL-4 promoter polymorphism C589T and the ADAM33 T1 polymorphism between asthmatic patients and controls (P = 0.638 and 0.943, respectively). Our data revealed that there is no association of these SNPs with asthma indicating that other SNPs of these genes or other genes might be involved in the manifestation of asthma.

Lack of association between asthma and ABO blood group.

ABO is the most important blood group system in transfusion and transplantation practices. Glycosyltransferases are controlled by the ABO system which is helpful in building oligosaccharide structures on the cell surface of erythrocytes and vascular endothelium and in the exocrine secretion system, including the respiratory tract. We analyzed the ABO blood group of 200 children and adults with asthma as well as that of 2000 healthy subjects as controls. The most common blood group among the patients and controls was "O" (43.5% and 43.6%, respectively), followed by B, A, and AB. In the distribution of different blood groups, nonsignificant difference between patients and controls was observed (p = 0.931). We conclude that ABO blood group status has a nonsignificant association with asthma among the population of Mysore, Karnataka, South India.

Soluble intercellular adhesion molecule-1 and E-selectin in patients with asthma exacerbation.

Soluble intercellular adhesion molecule-1 (sICAM-1) and soluble E-selectin (sE-selectin) are important factors in immunological processes of inflammatory cell buildup in target tissues. Studies have suggested that these molecules could be important markers of inflammatory diseases. This study was undertaken to assess the levels of sICAM-1 and sE-selectin during an acute attack of asthma in adults and children and to establish normal values (95th percentile) in healthy control subjects. We analyzed serum levels of ICAM-1 and E-selectin in 120 children and adults obtained during an acute attack of asthma: 40 with severe and 80 with moderately severe attack, and 50 healthy subjects as controls by ELISA (enzyme linked immunosorbent assay). sICAM-1 from patients with asthma was significantly higher than healthy controls (P < 0.001) but not for sE-selectin (P = 0.778). Significant differences were observed in moderately severe attack versus controls and severe attack of asthma for sICAM-1. With 95th percentile levels as cutoff for normal values (sICAM-1 = 585.08 ng/ml, sE-selectin = 160.87 ng/ml), it was observed that 88.3% of subjects (sICAM-1) and 98.5% of subjects (sE-selectin) with an acute attack of asthma had levels within the normal range. Although mean serum levels of sICAM-1 are higher in asthmatics than normal controls, it may be necessary to establish individual baseline values for serial estimation to evaluate their clinical relevance.