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