ORIGINAL_ARTICLE Determination of Haptoglobin Genotype in an Iranian Population with Idiopathic Generalized Epilepsy Background: Haptoglobin (Hp) is a plasma α2-sialoglycoprotein that contains alpha and beta chains. It displays in three common phenotypes, Hp1-1, Hp2-1, and Hp2-2. Proteins expressed by polymorphic genes have grossly different molecular sizes resulting in different diffusion rates in the brain. Haptoglobin expressed by the Hp2-2 genotype has lower hemoglobin-binding capacity than Hp1-1 or Hp2-1 and is associated with idiopathic generalized epilepsy. Methods: To determine polymorphism in haptoglobin genes in patients with idiopathic generalized tonic-clonic seizures, 42 men, 42 women, and 50 controls were selected for this study. Genomic DNA was extracted from blood and studied by polymerase chain reactions (PCR). Results: The amplified fragments for the Hp1-1 and Hp2-2 genotypes were 1757 and 3481 base pairs (bp) respectively, and the Hp2-1 genotype had both fragments, in addition to a 349-bp fragment. The distribution of the three major Hp phenotypes in epilepsy patients was 28.6 (1-1), 38.1 (2-1), and 33.3% (2-2) in the men, and 31 (1-1), 40.5 (2-1), and 28.6% (2-2) in the women. The distribution of Hp genotypes in controls was 22 (1-1), 40 (2-1), and 38% (2-2). Conclusion: We show that all Hp genotypes participate in idiopathic generalized epilepsy. http://rbmb.net/article-1-58-en.pdf 2015-04-30 51 55 Epilepsy Haptoglobin Iran Sukaina Al-balaghee 1 Department of Biology, Al Zahra University, Tehran, Iran AUTHOR Zeinab Al-balaghee 2 Department of Biology, Al Zahra University, Tehran, Iran AUTHOR Ashraf Shabani 3 Department of Biology, Al Zahra University, Tehran, Iran AUTHOR Parinaz Ghadam 4 Department of Biology, Al Zahra University, Tehran, Iran AUTHOR Mojgan Bandehpour 5 Cellular and Molecular Biology Research Centre, Shahid Beheshti University of Medical Sciences, Tehran, Iran - Biotechnology Department, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran AUTHOR Ali Askari Mehr 6 Iranian Epilepsy Association, Tehran, Iran AUTHOR Bahram Kazemi kazemi@sbmu.ac.ir 7 Cellular and Molecular Biology Research Centre, Shahid Beheshti University of Medical Sciences, Tehran, Iran - Biotechnology Department, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran AUTHOR
ORIGINAL_ARTICLE Biochemical Detection of N-Acyl Homoserine Lactone from Biofilm-Forming Uropathogenic Escherichia coli Isolated from Urinary Tract Infection Samples Background: N-Acyl homoserine lactone (AHL) is found to be the main component of quorum sensing (QS) in Gram-negative bacteria and plays an important role in biofilm formation. Little information is available regarding the role of AHL in biofilm formation in Escherichia coli (E. coli). The purpose of this investigation was to biochemically detect and characterize AHL activity in biofilm-forming uropathogenic E. coli (UPEC) isolated from urine samples of the patients with urinary tract infections (UTIs) in Kerman, Iran. Methods: Thirty-five UPEC isolates were obtained from urine samples of the patients with UTIs referred to the Afzalipoor hospital. The isolates were identified by biochemical tests. Biofilm analyses of all the isolates were performed using the microtiter plate method at OD 490nm. N-Acyl homoserine lactone was separated from cell mass supernatants by liquid-liquid extraction (LLE) and analyzed by a colorimetric method. N-Acyl homoserine lactone functional groups were identified by Fourier Transform-Infrared Spectroscopy (FT-IR). Results: The biofilm formation assay identified 10 (28.57%) isolates with strong, 16 (45.71%) with moderate, and 9 (25.71%) with weak biofilm activities. The UPEC isolates with strong and weak biofilm activities were subjected to AHL analyses. It was found that isolates with the highest AHL activities also exhibited strong adherence to microplate wells (P≤0.05). Two E. coli isolates with the highest AHL activities were selected for FT-IR spectroscopy. Peaks at 1764.33, 1377.99, and 1242.90 cm-1 correspond to the C=O bond of the lactone ring, and the N=H and C-O bonds of the acyl chain, respectively. Conclusion: We found that many UPEC isolates exhibited strong biofilm formation. The control of this property by AHL may contribute to the pathogenesis of the organism in UTI’s. http://rbmb.net/article-1-59-en.pdf 2015-04-30 56 61 Biofilm FT-IR N-acylhomoserine lactone Uropathogenic Escherichia coli Rohollah Taghadosi 1 Department of Microbiology and Virology, Kerman University of Medical Sciences, Kerman, Iran. AUTHOR Mohammad Reza Shakibaie mr_shakibaei@kmu.ac.ir 2 Department of Microbiology and Virology, Kerman University of Medical Sciences, Kerman, Iran - Research Center for Infectious and Tropical Diseases, Kerman University of Medical Sciences, Kerman, Iran. AUTHOR Shalaleh Masoumi 3 Department of Microbiology and Virology, Kerman University of Medical Sciences, Kerman, Iran. AUTHOR
ORIGINAL_ARTICLE Protective Effects of Withania somnifera Root on Inflammatory Markers and Insulin Resistance in Fructose-Fed Rats Background: We investigated the effects of Withania somnifera root (WS) on insulin resistance, tumor necrosis factor α (TNF-α), and interleukin-6 (IL-6) in fructose-fed rats. Methods: Forty-eight Wistar-Albino male rats were randomly divided into four groups (n=12); Group I as control, Group II as sham-treated with WS by 62.5mg/g per diet, Group III fructose-fed rats received 10%W/V fructose, and Group IV fructose- and WS-fed rats. After eight weeks blood samples were collected to measure glucose, insulin, IL-6, and TNF-α levels in sera. Results: Blood glucose, insulin, homeostasis model assessment for insulin resistance (HOMA-R), IL-6, and TNF-α levels were all significantly greater in the fructose-fed rats than in the controls. Treatment with WS significantly (P < 0.05) inhibited the fructose-induced increases in glucose, insulin, HOMA-R, IL-6, and TNF-α. Conclusion: Our data suggest that WS normalizes hyperglycemia in fructose-fed rats by reducing inflammatory markers and improving insulin sensitivity. http://rbmb.net/article-1-60-en.pdf 2015-04-30 62 67 IL-6 Insulin resistance TNF- α Withania somnifera Zahra Samadi Noshahr 1 Department of Pharmacology, Zahedan University of Medical Sciences, Zahedan, Iran AUTHOR Mohammad Reza Shahraki 2 Department of Physiology, Zahedan University of Medical Sciences, Zahedan, Iran AUTHOR Hassan Ahmadvand hassan_a46@yahoo.com 3 Department of Biochemistry, Faculty of Medicine, Lorestan University of Medical Sciences, Khorram Abad, Iran AUTHOR Davood Nourabadi 4 Department of Physiology, Iran University of Medical Sciences, Tehran, Iran AUTHOR Alireza Nakhaei 5 Department of Pharmacology, Zahedan University of Medical Sciences, Zahedan, Iran AUTHOR
ORIGINAL_ARTICLE The Anti-Proliferative and Anti-Angiogenic Effect of the Methanol Extract from Brittle Star Background: Anti-angiogenic therapy is a crucial step in cancer treatment. The discovery of new anti-angiogenic compounds from marine organisms has become an attractive concept in anti-cancer therapy. Because little data correlated to the pro- and anti-angiogenic efficacies of Ophiuroidea, which include brittle star, the current study was designed to explore the anti-angiogenic potential of brittle star methanol extract in vitro and in vivo. Methods: The anti-proliferative effect of brittle star extract on A2780cp cells was examined by MTT assays, and transcriptional expression of VEGF and b-FGF was evaluated by RT-PCR. In an in vivo model, 40 fertilized Ross eggs were divided into control and three experimental groups. The experimental groups were incubated with brittle star extract at concentrations of 25, 50 and 100 µg/ml, and photographed by photo-stereomicroscopy. Ultimately, numbers and lengths of vessels were measured by Image J software. Data were analyzed with SPSS software (p<0.05). Results: Results illustrated that the brittle star extract exerted a dose- and time-dependent anti-proliferative effect on A2780cp cancer cells. In addition, VEGF and b-FGF expression decreased with brittle star methanol extract treatment. Macroscopic evaluations revealed significant changes in the second and third experimental group compared to controls (p<0.05). Conclusion: These finding revealed the anti-angiogenic effects of brittle star methanol extract in vitro and in vivo confer novel insight into the application of natural marine products in angiogenesis-related pathologies. http://rbmb.net/article-1-61-en.pdf 2015-04-30 68 75 Angiogenesis Anti-proliferative Brittle star Marine organisms Ovarian cancer Javad Baharara baharara@yahoo.com 1 Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran AUTHOR Elaheh Amini 2 Faculty of Biological sciences, Developmental Biology, Kharazmi University, Tehran, Iran AUTHOR Marzieh Mousavi 3 Cell & Developmental Biology Research center for Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran AUTHOR
ORIGINAL_ARTICLE Expression of TGF-β3 in Isolated Fibroblasts from Foreskin Background: The multifunctional transforming growth factor beta (TGF-β) is a glycoprotein that exists in three isoforms. TGF-β3 expression increases in fetal wound healing and reduces fibronectin and collagen I and III deposition, and also improves the architecture of the neodermis which is a combination of blood vessels and connective tissue during wound healing. Fibroblasts are key cells in the wound healing process. TGF-β3 plays a critical role in scar-free wound healing and fibroblast actions in the wound healing process. The aim of this study was to express the TGF-β3 gene (tgf-b3) in human foreskin fibroblasts (HFF’s). Methods: We obtained HFF’s from a newborn and a primary fibroblast culture was prepared. The cells were transfected with TGF-β3-pCMV6-XL5 plasmid DNA by both lipofection and electroporation. Expression of TGF-β3 was measured by enzyme-linked immunosorbent assay (ELISA). Results: The highest TGF-β3 expression (8.3-fold greater than control) was obtained by lipofection after 72 hours using 3 µl of transfection reagent. Expression was 1.4-fold greater than control by electroporation. Conclusions: In this study, we successfully increased TGF-β3 expression in primary fibroblast cells. In the future, grafting these transfected fibroblasts onto wounds can help the healing process without scarring. http://rbmb.net/article-1-62-en.pdf 2015-04-30 76 81 Fibroblasts Gene expression TGF-β3 Mahnaz Mahmoudi Rad 1 Skin Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran AUTHOR Niki Mahmoudi Rad 2 Skin Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran AUTHOR Yasaman Mirdamadi yasaman.mirdamadi@med.ovgu.de 3 Skin Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran - Department of Dermatology and Venereology, Otto-von Guericke University Magdeburg, Magdeburg, Germany AUTHOR
ORIGINAL_ARTICLE Transforming Growth Factor Beta 1 869T/C and 915G/C Polymorphisms and Risk of Autism Spectrum Disorders Background: Transforming growth factor-β1 (TGF-β1) has been found to play a crucial role in early central nervous system development. Several studies have illustrated decreased TGF-β1 levels in sera and brains of autistic children. Two point mutations in the TGF-β1 signal peptide at 869T/C and 915G/C have been reported to influence TGF-β1 expression. The aim of the present study was to investigate the correlation of TGF-β1 polymorphisms and their haplotypes with autism. Methods: This study was performed on 39 autistic patients and 35 age- and sex-matched normal controls in an Iranian population, using the sequence specific primed-polymerase chain reaction (PCR-SSP) technique. Patients were divided into mild-to-moderate and severe groups according to the childhood autism rating scale. Results: No significant differences were observed for allele, genotype, or haplotype frequencies between the autistics and controls. Only a slight difference was observed in GC25 between the controls and all children with autism. Conclusion: Thus, these results indicate that the polymorphisms in TGF-β1 gene may not play an important role in the development of autism. http://rbmb.net/article-1-63-en.pdf 2015-04-30 82 88 Autism spectrum disorders Development Polymorphism Transforming Growth Factor beta 1 Mohammad Reza Khakzad mr.khakzad@gmail.com 1 Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran - Department of Immunology, Mashhad Branch, Islamic Azad University, Mashhad, Iran AUTHOR Farhad Salari 2 Immunology Research Center, Medical School, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Maryam Javanbakht 3 Department of Psychiatrics, Islamic Azad University, Mashhad Branch, Mashhad, Iran AUTHOR Maryam Hojati 4 Noor Hedayat Center of Autism Spectrum Disorders, Mashhad, Iran AUTHOR Abdolreza Varasteh 5 Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Mojtaba Sankian 6 Immunology Research Center, Medical School, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Mojtaba Meshkat 7 Department of Biostatistc, Shahid Beheshti University of medical sciences, Tehran, Iran AUTHOR
ORIGINAL_ARTICLE Pattern of Infection and Antibiotic Activity among Streptococcus agalactiae Isolates from Adults in Mashhad, Iran Background: One of the main causes of sexually transmitted diseases is group B β- hemolytic streptococci (GBS) multiplying in the genital tracts. Penicillin is the most common drug for the treatment of infections caused by these bacteria, but in patients suffering from Penicillin allergy, Erythromycin and Clindamycin are used as alternative therapeutic drugs against GBS. Recently, resistance to these drugs has been reported more often. In this study, efforts have been made to determine the prevalence and antibiotic resistance of GBS. Methods: Modified Christie Atkins Munch-Petersen (CAMP) test was conducted on over 2400 samples of urine and discharge taken from vagina, urethra and prostate. The drug sensitivity was performed by double disk sensitivity tests to Bacitracin, Trimethoprim, and Sulfamethoxazole and then the resistant samples were investigated by E-test to determine the minimal inhibitory concentrations (MICs) value. Results: Twenty-three vaginal and 10 urethral discharge, 27urine and 6 prostatic secretion samples were GBS positive. The most symbiotic microorganisms with GBS were strains of Enterococci (90%), Staphylococcus saprophyticus (25%) and Candida albicans (6%). The disk diffusion method showed 18 cases with Penicillin resistance (MIC: 1.5 mg/ml). Conclusion: Taken together, GBS carriers’ rate in this study was found 20.65% (8.24% men and 12.4% women). Furthermore, findings showed high-level resistance to Erythromycin and Clindamycin. http://rbmb.net/article-1-64-en.pdf 2015-04-30 89 93 Antibiotic resistance Genitourinary system Minimal inhibitory concentration (MIC) Streptococcus agalactiae Masoumeh Malek-Jafarian 1 Department of Laboratory Medicine, School of Paramedicine, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Fatemeh-Sadat Hosseini Hosseinif68@yahoo.com 2 Department of Laboratory Medicine, School of Paramedicine, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Abodol-Reza Ahmadi 3 Department of Laboratory Medicine, School of Paramedicine, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR