ORIGINAL_ARTICLE Methylmalonic Acidemia Diagnosis by Laboratory Methods Methylmalonic acidemia (MMA) is usually caused by a deficiency of the enzyme methylmalonyl-CoA mutase (MCM), a defect in the transport or synthesis of its cofactor, adenosyl-cobalamin (cblA, cblB, cblC, cblF, cblD, and cblX), or deficiency of the enzyme methylmalonyl-CoA epimerase. A comprehensive diagnostic approach involves investigations of metabolites with tandem mass spectrometry, organic acid analysis with gas chromatography, enzymatic studies with fibroblast cell culture, and finally, mutation analysis. With biochemical techniques and enzymatic assay the reliable characterization of patients with isolated MMA for mutation analysis can be achieved. Reliable classification of these patients is essential for ongoing and prospective studies on treatments, outcomes, and prenatal diagnoses. This article reviews the diagnostic techniques used to characterize patients with MMA. http://rbmb.net/article-1-79-en.pdf 2016-10-30 1 14 Diagnostic techniques Enzyme assay Methylmalonic acidemia Mutation analysis Organic acid analysis Tandem mass spectrometry Fatemeh Keyfi 1 Immunobiochemistry Lab, Immunology Research Center, School of medicine, Mashhad University of Medical Sciences, Mashhad, Iran - Pardis Clinical and Genetic Laboratory, Mashhad, Iran AUTHOR Saeed Talebi 2 Department of Medical Genetics, Faculty of Medicine, Tehran University of medical sciences, Tehran, Iran AUTHOR Abdol-Reza Varasteh varasteha@mums.ac.ir 3 Pardis Clinical and Genetic Laboratory, Mashhad, Iran - Immunobiochemistry Lab, Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran - Varastegan Institute for Medical Sciences, Mashhad, Iran. AUTHOR
ORIGINAL_ARTICLE Low Prevalence of Aeromonas hydrophilain Infectious Diarrhea Samples of Pediatric Patients in Arak, Iran Background: Aeromonashydrophila (A. hydrophila) is an aquatic bacterium that can cause a spectrum of infectious diseases, including both gastrointestinal and extraintestinal infections. Due to the high rate of diarrheal infections in pediatric patients in central Iran, this study was designed to determine the frequency of A. hydrophila in diarrhea samples from children in this region. Methods: In this descriptive cross-sectional study, diarrheal stool specimens were collected from 200 children admitted between February and October of 2015 to educational and medical centers affiliated with the Arak University of Medical Sciences. The samples were analyzed both phenotypically by culture and genotypically by the polymerase chain reaction (PCR). Results: A. hydrophila was isolated from two of the 200 samples tested (1%). The presence of bacterial genetic markers further confirmed the diagnosis. Conclusions: Based on this study, A. hydrophilais not highly prevalent in children with diarrhea in Arak; however clinical diagnostic laboratory personnel should be aware of the possible presence of A.hydrophila in children with diarrhea as it can cause dangerous health problems in both them and young adolescents. http://rbmb.net/article-1-80-en.pdf 2016-10-30 15 19 Aeromonas hydrophila Diarrhea Frequency Iran Pediatrics Elnaz Abbasi 1 Department of Microbiology & Immunology Faculty of Medicine Arak University of Medical Sciences, Arak, Iran AUTHOR Behzad Khansari-nejad 2 Department of Microbiology & Immunology Faculty of Medicine Arak University of Medical Sciences, Arak, Iran AUTHOR Hamid Abtahi 3 Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran AUTHOR Majid Akbari 4 Department of Microbiology & Immunology Faculty of Medicine Arak University of Medical Sciences, Arak, Iran AUTHOR Ehsanollah Ghaznavi-rad e.ghaznavirad@arakmu.ac.ir 5 Department of Microbiology & Immunology Faculty of Medicine Arak University of Medical Sciences, Arak, Iran - Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran AUTHOR
ORIGINAL_ARTICLE Trace Elements Status in Sera of Patients with Allergic Asthma Background: Asthma is a multifactorial disease and its severity varies with the inflammatory grade. There are conflicting reports about the roles of trace elements in asthma. This study examined the effects of zinc (Zn), copper (Cu), and selenium (Se) concentrations in sera of patients with allergic asthma attending Ghaem Hospital, Mashhad, Iran. Methods: Forty-nine patients, aged 10 to 50 years, with asthma in moderate or severe stages, and 24 healthy controls, were enrolled in this study. After demographic data collection and clinical evaluations, the subjects’ serum concentrations of Zn, Cu, and Se were measured via atomic absorbency. Results: Mean serum levels of Zn and Se in patients with allergic asthma were lower than in the healthy control group, but the Cu concentration in sera of patients with allergic asthma was slightly higher than healthy controls. Conclusions: Low levels of trace elements, specifically Zn, may have a role in the pathogenesis of allergic asthma; replacement of these elements may be an effective treatment. http://rbmb.net/article-1-81-en.pdf 2016-10-30 20 25 Asthma Hyper sensitivity Trace elements Nazila Ariaee 1 Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Reza Farid 2 Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Fahimeh Shabestari 3 Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Mohamad Shabestari 4 Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Farahzad Jabbari Azad Jabbarif@mums.ac.ir 5 Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR
ORIGINAL_ARTICLE Are Serum Levels of F2-Isoprostane and Oxidized-LDL Related to Vitamin D Status in Type 2 Diabetic Patients? A Case-Control Study Background: Considerable evidence suggests that oxidative stress affects diabetes mellitus (DM) and contributes to its complications. Vitamin D has been shown to possess antioxidant properties. The aim of this study was to determine the association between serum levels of calcifediol (25-OH-D), an indicator of vitamin D status, and lipid profiles with oxidative stress in patients with type 2 diabetes mellitus (T2DM). Methods: In this case-control study, 57 T2DM patients with low vitamin D status (< 30 ng/mL) and 48 T2DM patients with normal vitamin D status (> 30 ng/mL) were enrolled. Fasting concentrations of 25-OH-D, calcium, phosphorus, parathyroid hormone (PTH), lipid profiles, fasting blood sugar (FBS), glycosylated hemoglobin (HbA1c), F2-isoprostane, and oxidized-low-density lipoprotein (ox-LDL) were measured. Results: The mean fasting serum concentrations of 25-OH-D, calcium, and phosphorus in patients with low vitamin D status were significantly lower than in controls (p < 0.001). The mean concentrations of ox-LDL, F2-isoprostane, total cholesterol, and LDL were significantly higher in patients with low vitamin D status than in controls. There was a negative correlation between vitamin D levels and F2-isoprostane (r = 0.647and P = 0.0001), LDL (r = -0.218 and P = 0.030), and ox-LDL (r = -0.637 and P = 0.0001). Conclusions: The results of present study indicated that serum concentrations of 25-OH-D were inversely correlated with F2-isoprostane, LDL, and ox-LDL. Therefore, vitamin D may have a beneficial effect on the control of lipid profiles and oxidative stress in T2DM patients. http://rbmb.net/article-1-82-en.pdf 2016-10-30 26 32 Diabetes mellitus type 2 F2-isoprostane Oxidative stress Ox-LDL Vitamin D Mohammad Hassan Javanbakht 1 Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran. AUTHOR Hamed Mohammady 2 Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran. AUTHOR Koorosh Fooladsaz 3 Department of Clinical Biochemistry, Zanjan University of Medical Sciences, Zanjan, Iran AUTHOR Maryam Razzaghi 4 Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran. AUTHOR Mahnaz Zarei 5 Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran. AUTHOR Mahmoud Djalali mjalali87@yahoo.com 6 Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran. AUTHOR
ORIGINAL_ARTICLE Evaluation of SCD and FASN Gene Expression in Baluchi, Iran-Black, and Arman Sheep Background: With the increasing concern for health and nutrition, dietary fat has attracted considerable attention. The composition of fatty acids in the diet is important because they are associated with major diseases including cancers, diabetes, and cardiovascular disease. The fatty acid synthase (FASN) and stearoyl-CoA desaturase (delta-9-desaturase) (SCD) genes affect fatty acid composition (1). The expression of SCD and FASN genes is related to an increase in conjugated linoleic acid (CLA) in dairy products, which benefits human health.The aim of current study was to investigate expression changes of SCD and FASN genes that resulted from crossbreeding the local Baluchi sheep with alien breeds. Methods: We collected tissue samples from the mammary glands of 24 single-born ewes from local Baluchi and synthetic Iran-Black and Arman sheep breeds in the Abbas Abad breeding center. After RNA extraction and cDNA synthesis, real-time PCR was performed with all samples in triplicate. Results:The maximum and minimum expression of SCD and FASN genes was in the local Baluchi sheep and the crossbred Arman sheep, respectively. Conclusions: With the highest SCD and FASN gene expression in local Baluchi sheep and relatively less expression of these genes in synthetic Iran-Black and Arman Sheep breeds, it may be necessary to consider the consequences of crossbreeding local sheep and the fatty acid composition of their dairy products. http://rbmb.net/article-1-83-en.pdf 2016-10-30 33 39 SCD FASN Gene expression Real-time PCR Mohammad Salmani Izadi 1 Department of Animal Sciences, Ferdowsi University of Mashhad, International Campus, Mashhad, Iran AUTHOR Abbas Ali Naserian abasalin@yahoo.com 2 Department of Animal Sciences, Ferdowsi University of Mashhad, Mashhad, Iran. AUTHOR Mohammad Reza Nasiri 3 Department of Animal Sciences, Ferdowsi University of Mashhad, Mashhad, Iran. AUTHOR Reza Majidzadeh Heravi 4 Department of Animal Sciences, Ferdowsi University of Mashhad, Mashhad, Iran. AUTHOR Reza Valizadeh 5 Department of Animal Sciences, Ferdowsi University of Mashhad, Mashhad, Iran. AUTHOR
ORIGINAL_ARTICLE Quantification of Pla or 3, a Platanus orientalis Allergen, Grown under Different Environmental Conditions, by Sandwich ELISA Background: Platanus species are widely cultured around the world and considered an important cause of allergic reactions. In the present study, we developed a sandwich ELISA to quantify Pla or 3 allergen in P. orientalis pollen extracts grown near high-traffic roads and compared it to pollen extracts collected from rural areas as control. Methods: Pollen samples were collected from three polluted and two unpolluted sites in Mashhad, northeast Iran. Recombinant Pla or 3 was expressed and used for polyclonal antibody production in rabbit. A sandwich ELISA was developed and validated to quantify Pla or 3 levels in pollen extracts from the different sites. Results: The coefficients of variation (CVs) for the intra- and inter- day assays were less than 5 and 18%, respectively. The working range of the standard curve was between 0.1 and 25 ng/ml, with the detection limit being 0.037 ng/ml. The recovery percentage was 88-106.4% at working concentrations from 0.31 to 26.5 ng/ml. Pla or 3 levels were significantly greater in pollens grown near high-traffic roads than in those grown in rural regions (p < 0.0001). Conclusions: A sandwich ELISA was developed and validated to quantify Pla or 3 in pollen extracts. Using this validated ELISA, we showed a substantial difference between the amounts of Pla or 3 in pollens grown in different environments. This finding should be considered in developing public policies to reduce traffic pollution, which leads to reduced allergic reactions in atopic subjects. http://rbmb.net/article-1-84-en.pdf 2016-10-30 40 45 Pathogenesis-related proteins Pla or 3 Platanus orientalis Sandwich ELISA Farnaz Sedghy 1 Immuno-Biochemistry lab, Immunology Research Center, Mashhad University of medical Sciences, Mashhad, Iran AUTHOR Mojtaba Sankian 2 Immuno-Biochemistry lab, Immunology Research Center, Mashhad University of medical Sciences, Mashhad, Iran AUTHOR Maliheh Moghadam 3 Immuno-Biochemistry lab, Immunology Research Center, Mashhad University of medical Sciences, Mashhad, Iran AUTHOR Abdol-Reza Varasteh varasteha@mums.ac.ir 4 Allergy Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR
ORIGINAL_ARTICLE Design and Construction of a Cloning Vector Containing the hspX Gene of Mycobacterium tuberculosis Background: Tuberculosis (TB) is a major cause of death worldwide. Finding an effective vaccine against TB is the best way to control it. Several vaccines against this disease have been developed but none are completely protective. The aim of this study was to design and construct a cloning vector containing the Mycobacterium tuberculosis (M. tuberculosis) heat shock protein X (hspX). Methods: First, an hspX fragment was amplified by PCR and cloned into plasmid pcDNA3.1(+) and recombinant vector was confirmed. Results: A 435 bp hspX fragment was isolated. The fragment was 100% homologous with hspX of M. tuberculosis strain H37Rv in GenBank. Conclusions: In this study, the cloning vector pcDNA3.1(+), containing a 435-bp hspX fragment of M. tuberculosis, was constructed. This could be used as a DNA vaccine to induce immune responses in animal models in future studies. http://rbmb.net/article-1-85-en.pdf 2016-10-30 46 50 Cloning DNA vaccine hspX Mycobacterium tuberculosis Atieh Yaghoubi 1 Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Ehsan Aryan 2 Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Hosna Zare 3 Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Shadi Alami 4 Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Roghayeh Teimourpour 5 Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran. AUTHOR Zahra Meshkat meshkatz@mums.ac.ir 6 Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR
ORIGINAL_ARTICLE Association of FAS A-670G Polymorphism and Risk of Uterine Leiomyoma in a Southeast Iranian Population Background: Uterine leiomyoma (UL) is a benign tumor of uterine smooth muscle that affects women in reproductive ages. FAS has an important role in initial stages of apoptosis. Previous studies have shown an association between the FAS gene and tumorigenesis. In the present study, we evaluated the relationship between FAS A-670G (rs 1800682) and UL risk. Methods: The FAS gene polymorphism of 155 women with UL and 157 healthy controls was analyzed by the polymerase chain reaction restriction fragment length polymorphism method. Results: The AA, AG, and GG genotype frequencies of the FAS A-670G polymorphism were respectively 37.4, 42.6, and 20% in women with UL, and 46, 42.6, and 11.5% in healthy controls. The risk of UL in women was 1.5-fold greater in GG-genotype women than in AA-genotype women. The G allele frequencies were 41% in women with UL and 33% in healthy controls and statistically different (P = 0.03). Conclusions: The FAS polymorphism was associated with the risk of UL in a sample of Iranian women. http://rbmb.net/article-1-86-en.pdf 2016-10-30 51 55 FAS PCR-RFLP Polymorphism Uterine leiomyoma Abbas Mohammadpour-Gharehbagh 1 Student Scientific Research Center, Zahedan University of Medical Sciences, Zahedan, Iran - Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran. AUTHOR Saeedeh Salimi sasalimi@yahoo.com 2 Cellular and Molecular Research Center, Zahedan University of Medical Sciences, Zahedan, Iran - Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran. AUTHOR Farshid Keshavarzi 3 Cellular and Molecular Research Center, Zahedan University of Medical Sciences, Zahedan, Iran - Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran. AUTHOR Sepideh Zakerian 4 Department of Obstetrics and Gynecology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran. AUTHOR Mojtaba Sajadian 5 Cellular and Molecular Research Center, Zahedan University of Medical Sciences, Zahedan, Iran - Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran. AUTHOR Mojgan Mokhtari 6 Department of Obstetrics and Gynecology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran. AUTHOR
ORIGINAL_ARTICLE Establishment a CHO Cell Line Expressing Human CD52 Molecule Background: CD52 is a small glycoprotein with a GPI anchor at its C-terminus. CD52 is expressed by Normal and malignant T and B lymphocytes and monocytes. There are detectable amounts of soluble CD52 in plasma of patients with CLL and could be used as a tumor marker. Although the biological function of CD52 is unknown but it seems that CD52 may be involved in migration and activation of T-cells .The aim of this study was to clone and express human CD52 gene in CHO cell line and studying its function in more details. Methods: Based on GenBank databases two specific primers were designed for amplification of cd52 gene. Total RNA was extracted from Raji cell line and cDNA synthesized. Amplified fragment was cloned in pBudCE4.1 vector. The new construct was transfected to CHO-K1 cell line using electroporation method. Expression of recombinant CD52 protein was evaluated by Real time PCR and flow cytometry methods. Results: Amplification of CD52 gene using specific primers on Raji cDNA showed a 209 bp band. New construct was confirmed by PCR and restriction pattern and sequence analysis. The new construct was designated as pBudKT1. RT-PCR analysis detected cd52 mRNAs in transfected cells and Flow cytometry Results showed that 78.4 % of cells represented CD52 in their surfaces. Conclusions: In conclusion, we established a human CD52 positive cell line, CHO-CD52, and the protein was expressed on the membrane. Cloning of the CD52 gene could be the first step for the production of therapeutic monoclonal antibodies and detection systems for soluble CD52 in biological fluids. http://rbmb.net/article-1-87-en.pdf 2016-10-30 56 61 CD52 Recombinant DNA Therapeutic and diagnostic proteins Khadijeh Tati 1 Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran - Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran AUTHOR Mahsa Yazdanpanah-Samani 2 Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran AUTHOR Amin Ramezani 3 Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran - Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran AUTHOR Elham Mahmoudi Maymand 4 Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran AUTHOR Abbas Ghaderi ghaderia@sums.ac.ir 5 Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran AUTHOR
ORIGINAL_ARTICLE Molecular Analysis and Expression of bap Gene in Biofilm-Forming Multi-Drug-Resistant Acinetobacter baumannii Background: Acinetobacter baumannii is commonly resistant to nearly all antibiotics due to presence of antibiotic resistance genes and biofilm formation. In this study we determined the presence of certain antibiotic-resistance genes associated with biofilm production and the influence of low iron concentration on expression of the biofilm-associated protein gene (bap) in development of biofilm among multi-drug-resistant A. baumannii (MDRAB). Methods: Sixty-five MDRAB isolates from clinical samples were collected. Molecular typing was carried out by random amplified polymorphism DNA polymerase chain reaction (RAPD-PCR). Biofilm formation was assayed by the microtiter method. Results: The sequence of bap was determined and deposited in the GenBank database (accession no. KR080550.1). Expression of bap in the presence of low iron was analyzed by relative quantitative real time PCR (rqRT-PCR). Nearly half of the isolates belonged to RAPD-types A and B remaining were either small clusters or singleton. The results of biofilm formation revealed that 23 (35.4%), 18 (27.7%), 13 (20%), and 11 (16.9%) of the isolates had strong, moderate, weak, and no biofilm activities, respectively. ompA and csuE genes were detected in all, while bap and blaPER-1 were detected in 43 (66%) and 42 (64%) of the isolates that showed strong and moderate biofilm activities (p ≤ 0.05), respectively. Analysis of bap expression by rqRT-PCR revealed five isolates with four-fold bap overexpression in the presence of low iron concentration (20 μM). Conclusion: The results suggest that bap overexpression may influence biofilm formation in presence of low iron concentration. http://rbmb.net/article-1-88-en.pdf 2016-10-30 62 72 Acinetobacter baumannii Biofilm Biofilm-associated Protein (bap) Iron rqRT-PCR Omid Azizi 1 Department of Microbiology and Virology, Kerman University of Medical Sciences, Kerman, Iran AUTHOR Fereshteh Shahcheraghi 2 Department of Bacteriology, Pasteur institute of Iran, Tehran, Iran AUTHOR Himen Salimizand 3 Department of Microbiology and Virology, Kurdistan University of Medical Sciences, Kurdistan, Iran AUTHOR Farzan Modarresi 4 Department of Microbiology, School of Medicine, Jahrom University of Medical Sciences, Jahrom, Iran AUTHOR Mohammad Reza Shakibaie mr_shakibaei@kmu.ac.ir 5 Department of Microbiology and Virology, Kerman University of Medical Sciences, Kerman, Iran - Environmental Health Sciences and Engineering Research Center; Kerman University of Medical Sciences, Kerman, Iran - Research Center for Infectious Diseases and Tropical Medicine, Kerman University of Medical Sciences AUTHOR Shahla Mansouri 6 Department of Microbiology and Virology, Kerman University of Medical Sciences, Kerman, Iran AUTHOR Rashid Ramazanzadeh 7 Department of Bacteriology, Pasteur institute of Iran, Tehran, Iran - Cellular & Molecular Research Center and Microbiology Department, Faculty of Medicine, Kurdistan University of Medical Sciences, Iran AUTHOR Farzad Badmasti 8 Department of Bacteriology, Pasteur institute of Iran, Tehran, Iran AUTHOR Vajihe Nikbin 9 AUTHOR