@article{ author = {Abolhassani, Mohse}, title = {Obituary: Professor Fereydoun Malekzadeh (1933-2012)}, abstract ={We were greatly saddened to hear the news of Professor Fereydoun Malekzadeh's death at age of 79 in September 9, 2012. Professor Malekzadeh was a distinguished and highly respected Iranian Microbiologist and a member of the editorial board of the Iranian Biomedical Journal since October 1997. He was born in 1933 in city of Tabriz in Province of Azarbaijan, Iran. After graduated from high school, he moved to Tehran and attended at Tehran University and obtained his BS in Biology in 1956 and subsequently his MSc degree in Mycology in 1959. Professor Malekzadeh received “Fulbright Scholarship” and obtained his Ph.D. degree in Bacteriology in 1961 from Louisiana State University. He returned home and since then taught microbiology in Biology Department of Faculty of Science, in Tehran University and later in Azad University. His contribution to microbiology was immense and he played a major role in establishing the Doctoral degree in the Microbiology Department of Faculty of Science in Tehran University and in Azad University. He was also a key funder of Iranian Society for Biology and a member of "American Society for Microbiology". Dr. Malekzadeh wrote several reference books, such as Introduction to Microbiology, Medical Microbiology, Infectious Diseases, etc., and also authored several papers in international peer-reviewed journals. His many academic awards included two best scientific books of the year, Botany and Introduction to Microbiology co-authored with Dr. Moghaddam and Dr. Shahamat, respectively. He also received "UNESCO Science Prize" in 1999, and "International Kharazmi Award" for discovering two new strains of bacteria, "Cellulomonas Persia" and "Cellulomonas iranesis" that have been established in Bergey's Manual of Systematic Bacteriology and deposited in ATCC. Dr. Malekzadeh was visiting professor at University of Illinois, State University of Arizona, University of Göttingen, University of Alberta and Institute of Marine Biotechnology in University of Maryland. Dr. Malekzadeh loved to work at the bench and continued to do so long after his official retirement. He was extraordinarily hard-working, and held very high standards not only in his work but also in his personal life. He loved interacting with young people. He also spotted talent, and went to extraordinary lengths to promote young, talented scientists. Professor Malekzadeh was diagnosed with a neurodegenerative disease in 2006, which caused deterioration in his health over a long period of time. He demonstrated great courage, determination and dignity during this very difficult time, seeming more concerned about others than about himself. He showed clearly strength of character during his illness. He is survived by his wife and two daughters, Dr. Shirin and Katayoun Malekzadeh.}, Keywords = {}, volume = {17}, Number = {1}, pages = {0-0}, publisher = {Pasteur Institute of Iran}, url = {http://ibj.pasteur.ac.ir/article-1-936-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-936-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Tanomand, Asghar and Farajnia, Safar and Najar, Shahin and Majidi, Jafar}, title = {Cloning, Expression and Characterization of Recombinant Exotoxin A-Flagellin Fusion Protein as a New Vaccine Candidate against Pseudomonas aeruginosa Infections}, abstract ={Background: Infections due to Pseudomonas aeruginosa are among the leading causes of morbidity and mortality in patients who suffer from impaired immune responses and chronic diseases such as cystic fibrosis. At present, aggressive antibiotic therapy is the only choice for management of P. aeruginosa infections, but emergence of highly resistant strains necessitated the development of novel alternative therapeutics including an effective vaccine. Several P. aeruginosa antigens have been tested for vaccine development, including lipopolysaccharide alone, polysaccharides alginate, extracellular proteins, exotoxin A (exo A) and killed whole cell. However, none of them are currently available clinically. Methods: In this research, recombinant exoA-flagellin (fliC) fusion protein as a cocktail antigen was expressed, purified and its antigenic characteristics were evaluated. Results: Expression of recombinant fusion protein by E. coli using pET22b vector resulted in production of exoA-fliC fusion protein in high concentration. Based on Western-blotting results, recombinant fusion protein showed a good antigenic interaction with sera from patients with various P. aeruginosa infections. Conclusion: These results suggested that recombinant exoA-fliC fusion protein can be produced in the laboratory, and tested as a candidate vaccine in P. aeruginosa infections.}, Keywords = {Pseudomonas aeruginosa, Exotoxin A (exoA), Flagellin (fliC), Vaccines}, volume = {17}, Number = {1}, pages = {1-7}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1122.2012}, url = {http://ibj.pasteur.ac.ir/article-1-898-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-898-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Darabi, Shahram and Tiraihi, Taki and Delshad, AliReza and Sadeghizadeh, Maji}, title = {A New Multistep Induction Protocol for the Transdifferentiation of Bone marrow Stromal Stem Cells into GABAergic Neuron-Like Cells}, abstract ={Background: Bone marrow stromal stem cells (BMSC) are appropriate source of multipotent stem cells that are ideally suited for use in various cell-based therapies. It can be differentiated into neuronal-like cells under appropriate conditions. This study examined the effectiveness of co-stimulation of creatine and retinoic acid in increasing the differentiation of BMSC into GABAergic neuron-like cells (GNLC). Methods: BMSC isolated from the femurs and tibias of adult rats were cultured in DMEM/F12 medium supplemented with 10% FBS, pre-induced using β-mercaptoethanol (βME) and induced using retinoic acid (RA) and creatine. Immunostaining of neurofilament 200 kDa, neurofilament 160 kDa, nestin, fibronectin, Gamma-amino butyric acid (GABA) and glutamic acid decarboxylase (GAD) 65/67 were used to evaluate the transdifferentiation of BMSC into GNLC and to evaluate the effectiveness of pre-induction and induction assays. The expression of genes that encode fibronectin, octamer-binding transcription factor 4 (Oct-4), GAD 65/67 and the vesicular GABA transporter was examined in BMSC and GNLC by using RT-PCR assays during transdifferentiation of BMSC into GLNC. Results: Co-stimulation with RA and creatine during the induction stage doubled the rates of GABAergic differentiation compared with induction using creatine alone, resulting in a 71.6% yield for GLNC. RT-PCR showed no expression of Oct-4 and fibronectin after the induction stage. Conclusion: The results of this study showed that the application of βME, RA, and creatine induced the transdifferentiation of BMSC into GLNC.}, Keywords = {Cell therapy, GABAergic neurons, Creatine, Bone marrow stem cell}, volume = {17}, Number = {1}, pages = {8-14}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/IBJ.1112.2012}, url = {http://ibj.pasteur.ac.ir/article-1-812-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-812-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Yadegarazari, Reza and Hassanzadeh, Taghi and Majlesi, Amir and Keshvari, Amir and MonsefEsfahani, Alireza and Tootoonchi, Amirsasan and Shabab, Nooshin and Saidijam, Massou}, title = {Improved Real-Time RT-PCR Assays of Two Colorectal Cancer Peripheral Blood mRNA Biomarkers: A Pilot Study}, abstract ={Background: Efficient screening for detection of colorectal cancer (CRC) at earlier stages reduces its mortality. The purpose of this study was to investigate expression of carcinoembryonic antigen (CEA) and human telomerase reverse transcriptase (hTERT) mRNA in peripheral blood of CRC patients and to present strategies for early detection screen test. Methods: Twenty seven patients in non-metastatic stage and 27 healthy individuals were studied. Expression of CEA, hTERT mRNA and 18srRNA (18s subunit of ribosomal RNA, as reference gene) were determined based on real-time RT-PCR on 3 µg of total RNA from blood in 3 separate vials (1 µg per vial). Results: Positive expression rate of CEA mRNA (78%) and hTERT mRNA (81%) were higher in patient group (P<0.001). These rates were meaningfully higher than the results of individual vials containing only 1 µg of total RNA. Difference between Ct values of markers with 18srRNA (ΔCt) was higher in healthy group than patient one. Therefore, a ΔCt cut-off value was determined for distinguishing between true- and false-positive results. Concurrent expression of both markers was found in 67% of the patients, which was higher than healthy cases (11%). Combination of concurrent marker expression with cut-off point strategy increased specificity to 100%. Conclusion: These results showed that concurrent evaluation of marker expression and performing the test on 3 µg of samples in 3 separate vials may increase specificity and sensitivity of real-time RT-PCR for early detection of non-metastatic CRC. However, more investigations with larger numbers of samples are needed to verify these results.}, Keywords = {Carcinoembryonic antigen, Biomarker, Colorectal cancer}, volume = {17}, Number = {1}, pages = {15-21}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/IBJ.1104.2012}, url = {http://ibj.pasteur.ac.ir/article-1-809-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-809-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {SamsamShariat, Seyed Ziyae Aldin and Mostafavi, Sayed Abolfazl and Khakpour, Farz}, title = {Antioxidant Effects of Vitamins C and E on the Low-Density Lipoprotein Oxidation Mediated by Myeloperoxidase}, abstract ={Background: Oxidative modification of low-density lipoprotein (LDL) appears to be an early step in the pathogenesis of atherosclerosis. Meanwhile, myeloperoxidase (MPO)-catalyzed reaction is one of the potent pathway for LDL oxidation in vivo. The aim of this study was to evaluate in vitro antioxidant effects of vitamins C and E on LDL oxidation mediated by MPO. Methods: MPO was isolated from fresh plasma by sequential centrifugation using density ultracentrifugation. It was incubated with LDL and the LDL oxidation level was determined spectrophotometrically by measuring conjugated diene absorbance at 234 nm. Furthermore, vitamin C (50-200 mM) and vitamin E (10-40 mM) were added and the LDL oxidation level was determined. Results: The purity index of MPO and its enzymatic activity were 0.69 and 1127 U/mg protein, respectively. It was demonstrated that vitamin C in vitro inhibited LDL oxidation mediated by MPO however, vitamin E was unable to act in the same way. The protection by vitamin C was concentration dependent and maximum protective effect of vitamin C was observed at 150 mM, where about 64% of the LDL oxidation was inhibited. Vitamin C increased lag time of LDL oxidation mediated by MPO up to 2.4 times. Conclusion: It can be concluded from our results that vitamin C is able to improve LDL resistance to oxidative modification in vitro. In addition, vitamin C might be effective in LDL oxidation mediated by MPO in vivo, resulting in reduction of atherosclerosis process rate.}, Keywords = {Antioxidant, Myeloperoxidase (MPO), Low-density lipoprotein (LDL), Vitamin E}, volume = {17}, Number = {1}, pages = {22-28}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1092.2012}, url = {http://ibj.pasteur.ac.ir/article-1-798-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-798-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {BinAleem, Shoaib and Hussain, Muhammad Mazhar and Farooq, Yasir}, title = {Levo-Carnitine Reduces Oxidative Stress and Improves Contractile Functions of Fast Muscles in Type 2 Diabetic Rats}, abstract ={Background: Metabolic derangements in type 2 diabetes mellitus (T2DM) are likely to affect skeletal muscle contractile functions adversely. Levo-carnitine improves muscle contractile functions in healthy humans and rats and corrects metabolic derangements in T2DM. Therefore, it is likely to improve muscle contractile functions in T2DM as well. This study was designed to determine the effect of levo-carnitine on serum levo-carnitine levels, oxidative stress and contractile parameters of fast muscle in T2DM. Methods: Ninety Sprague-Dawley rats were randomly divided into three equal groups. Healthy rats served as the controls, while T2DM was induced in diabetic and carnitine groups. The carnitine group was administered levo-carnitine 200 mg/kg/day intraperitoneally for 6 days. At 28th day, extensor digitorum longus muscles were removed and their functions were assessed using iWorx data acquisition unit (AHK/214). Blood obtained by intra-cardiac sampling at 28th day was used for estimation of serum malondialdehyde (MDA) and levo-carnitine levels. Results: Maximum isometric twitch tension, time-to-peak twitch tension and time-to-relax to 50% of the peak twitch tension were not significantly different amongst the groups. Carnitine group showed significant improvement in maximum fused tetanic tension, maximum fused tetanic tension after fatigue protocol and recovery from fatigue after 5 minutes of rest period compared to the diabetic group. Serum MDA levels were reduced, while serum levo-carnitine levels were elevated significantly in carnitine group as compared to the diabetic group. Conclusion: Levo-carnitine supplementation increases serum levo-carnitine levels which decreases oxidative stress. This action improves contractile force but delays fatigue in fast muscles of diabetic rats.}, Keywords = {Type 2 diabetes mellitus (T2DM), Levo-carnitine, Fast muscles, Contractile functions, Oxidative stress}, volume = {17}, Number = {1}, pages = {29-35}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1086.2012}, url = {http://ibj.pasteur.ac.ir/article-1-796-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-796-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Tajalli, Raziye and Nobakht, Maliheh and Mohammadi-Barzelighi, Hajar and Agah, Shahram and Rastegar-Lari, Abdolaziz and Sadeghipour, Alirez}, title = {The Immunohistochemistry and Toluidine Blue Roles for Helicobacter pylori Detection in Patients with Gastritis}, abstract ={Background: Helicobacter pylori, which is associated with many upper gastrointestinal diseases, is found in half of the population of the world. Several special stains and immunohistochemistry stain for H. pylori are available. The need for and usefulness of immunohistochemical (IHC) technique has been debated for years. Toluidine blue is a simple stain for microbiological studies and is easily available in laboratories. Therefore, this study was conducted to compare hematoxylin and eosin (H&E), Giemsa and toluidine blue staining with immunehistochemistry for detection of H. pylori in patients with gastritis and also to correlate the results of these staining methods with pathological grading. Methods: We reviewed 54 consecutive gastric biopsy specimens stained by H&E and Giemsa as well as by toluidine blue and immunohistochemistry stains for H. pylori. Results: H. pylori was positively identified by IHC in 43 (79.63%) patients, while positive samples were found in 18 (33.33%), 24 (44.44%) and 33 (61.11%) patients using H&E, Giemsa and toluidine blue staining methods. Our results showed that classical histological staining methods are not sensitive enough to identify low numbers or coccoid forms of organism, while toluidine blue and immunohistochemistry play an important role in detection of H. pylori infection. Conclusion: Toluidine blue has been proved to be much more reliable than H&E and Giemsa in detection of H. pylori. In addition, in post treatment biopsies and in biopsies with unexplained chronic active gastritis without histological evidence of H. pylori should have immunohistochemistry done to detect possible low density or coccoid form of organisms.}, Keywords = {Helicobacter pylori, Immunohistochemistry, Gastritis}, volume = {17}, Number = {1}, pages = {36-41}, publisher = {Pasteur Institute of Iran}, doi = { 10.6091/IBJ.1094.2012}, url = {http://ibj.pasteur.ac.ir/article-1-810-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-810-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Taghizadeh, Mohsen and Talaei, Sayyed Alireza and Salami, Mahmou}, title = {Vitamin D Deficiency Impairs Spatial Learning in Adult Rats}, abstract ={Background: Through its membrane and intracellular receptors, vitamin D regulates many vital functions in the body including its well known actions on musculoskeletal system. Growing body of evidences demonstrate that vitamin D undergoes some of behavioral aspects of neurocognition. The present study was designed to evaluate the effect of food regimens without vitamin D or with a supplement of 1,25(OH)2D3 on spatial performance of adult rats. Methods: The animals were trained in the Morris water maze to find a hidden platform. The time spent and the distance travelled to find the platform, speed of navigation and the percentage of unsuccessful trials were considered for assessment of the task learning. Results: Our findings indicated that the vitamin D-deprived rats had a significant lower performance compared to both the controls and the animals receiving 1,25(OH)2D3 supplementation. Concerning the unsuccessful trials, lack of vitamin D resulted in the highest failures in the maze navigation. The regimen with additional 1,25(OH)2D3 did not considerably influence learning of the maze task. Conclusion: We concluded that while vitamin D deficiency deteriorates the spatial task learning, the 1,25(OH)2D3 supplementation did not effectively underlie the maze performance.}, Keywords = {Vitamin D, Maze learning, Dietary supplements}, volume = {17}, Number = {1}, pages = {42-48}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1061.2012}, url = {http://ibj.pasteur.ac.ir/article-1-795-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-795-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Yousefvand, Namdar and Zarei, Fatemeh and Ghanbari, Ali}, title = {Exogenous Testosterone, Finasteride and Castration Effects on Testosterone, Insulin, Zinc and Chromium in Adult Male Rats}, abstract ={Background: Although effects of trace elements on secretion of sex steroids and insulin have been studied, the effects of these hormones on serum level of trace elements have been rarely investigated. The aim of the present study was to evaluate the effect of testosterone and finasteride administration and castration on serum levels of testosterone, insulin, zinc and chromium. Methods: Male adult rats (n = 32) were divided into 4 groups (n = 8). Group 1, control Group 2, castration, castration was done at the first day of the study Group 3, finasteride (20 mg/kg/day, dissolved in drinking water) and Group 4, testosterone (5 mg/kg/day, i.p.). At the end of the period of the study (35 days), serum testosterone, insulin, zinc and chromium levels were determined in the blood samples collected directly from the right atrium of the heart of the animals. Results: The data indicated that the serum levels of testosterone, insulin and zinc were significantly increased (P<0.01) in testosterone-administrated and finasteride groups, but the level of chromium was decreased in both groups (P<0.01). Castrated group had the lowest serum levels of testosterone, insulin and zinc (P<0.05). Also, the levels of serum chromium in this group were increased. Conclusion: The study demonstrates that testosterone and finasteride increases insulin and zinc levels and decreases chromium levels in the serum of male adult rats. According to these data, it seems that testosterone may affect glucose cycle through effect on serum insulin levels and trace elements such as zinc and chromium.}, Keywords = {Finasteride, Castration, Insulin, Zinc, Chromium}, volume = {17}, Number = {1}, pages = {49-53}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1110.2012}, url = {http://ibj.pasteur.ac.ir/article-1-901-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-901-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Alikhani, Mehdi and SharifiTabar, Mehdi and Mirshahvaladi, Shahab and Kheimeh, Abolfazl and SadighiGilani, Mohammad Ali and Sabbaghian, Marj}, title = {Expression Analysis of RNA-Binding Motif Gene on Y Chromosome (RBMY) Protein Isoforms in Testis Tissue and a Testicular Germ Cell Cancer-Derived Cell Line (NT2)}, abstract ={a key factor in spermatogenesis and disorders associated with this protein have been recognized to be related to male infertility. Although it was suggested that this protein could have different functions during germ cell development, no studies have been conducted to uncover the mechanism of this potential function yet. Here, we analyzed the expression pattern of RBMY protein isoforms in testis compared to NT2, a testicular germ cell cancer-derived cell line, to test probability of differential expression of RBMY protein isoforms at different spermatogenesis stages. Methods: Full length and a segment of RBMY gene were cloned and expressed in E. coli. Anti-human RBMY antibody was produced in rabbit using the recombinant proteins as antigen. Western-blot and immunofluorescence were conducted for detection and comparison of RBMY protein isoforms. Results: Selected segment of RBMY protein resulted in producing a mono-specific antibody. As results shows, only the longest isoform of RBMY was expressed at protein level in NT2 cell line, while three isoforms of this protein were detected in the whole testis lysate. Conclusion: The results imply that different alternative splicing may happen in testis cells and probably difference of RBMY function during spermatogenesis is due to the differential expression of RBMY protein isoforms. These results and further experiments on RBMY isoforms can help to obtain a better understanding of the function of this protein, which may increase our knowledge about spermatogenesis and causes of male infertility.}, Keywords = {Protein isoforms, Spermatogenesis, Male infertility}, volume = {17}, Number = {2}, pages = {54-61}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1148.2013}, url = {http://ibj.pasteur.ac.ir/article-1-944-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-944-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Abbaszadeh, Hojjat-Allah and Tiraihi, Taki and Delshad, Ali Reza and SaghediZadeh, Majid and Taheri, Taher}, title = {Bone Marrow Stromal Cell Transdifferentiation into Oligodendrocyte-Like Cells Using Triiodothyronine as a Inducer with Expression of Platelet-Derived Growth Factor α as a Maturity Marker}, abstract ={Background: The present study investigated the functional maturity of oligodendrocyte derived from rat bone marrow stromal cells (BMSC). Methods: The BMSC were isolated from female Sprague-Dawley rats and evaluated for different markers, such as fibronectin, CD106, CD90, Oct-4 and CD45. Transdifferentiation of OLC from BMSC was obtained by exposing the BMSC to DMSO and 1 µM all-trans-retinoic acid during the pre-induction stage and then induced by heregulin (HRG), platelet-derived growth factor AA (PDGFR-α), fibroblast growth factor and T3. The neuroprogenitor cells (NPC) were evaluated for nestin, neurofilament 68, neurofilament 160 and glial fibrillary acidic protein gene expression using immunocytochemistry. The OLC were assessed by immunocytochemistry for O4, oligo2, O1 and MBP marker and gene expression of PDGFR-α was examined by RT-PCR. Results: Our results showed that the fibronectin, CD106, CD90, CD45 and Oct-4 were expressed after the fourth passage. Also, the yield of OLC differentiation was about 71% when using the O1, O4 and oligo2 markers. Likewise, the expression of PDGFR-α in pre-oligodendrocytes was noticed, while MBP expression was detected in oligodendrocyte after 6 days of the induction. Conclusion: The conclusion of the study showed that BMSC can be induced to transdifferentiate into mature OLC.}, Keywords = {Bone marrow stromal cell, Triiodothyronine, Platelet-derived growth factor α}, volume = {17}, Number = {2}, pages = {62-70}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.11162.2013 }, url = {http://ibj.pasteur.ac.ir/article-1-910-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-910-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Albukhaty, Salim and Naderi-Manesh, Hossein and Tiraihi, Taki}, title = {In vitro Labeling of Neural Stem Cells with Poly-L-Lysine Coated Super Paramagnetic Nanoparticles for Green Fluorescent Protein Transfection}, abstract ={Background: The magnetic nanoparticle-based transfection method is a relatively new technique for delivery of functional genes to target tissues. We aimed to evaluate the transfection efficiency of rat neural stem cell (NSC) using poly-L-lysine hydrobromide (PLL)-coated super paramagnetic iron oxide nanoparticles (SPION). Methods: The SPION was prepared and coated with PLL as transfection agent and the transfection efficiency was evaluated in rat NSC using enhanced green fluorescent protein-N1 plasmid containing GFP as a reporter gene. NSC was incubated for 24 h in cell culture media containing 25 µg/ml SPION and in different concentrations of PLL (0.25, 0.50, 0.75, 1 and 2 µg/ml). Cell viability was determined before and after transfection for every concentration using Trypan blue assay. Characterization of prepared uncoated (SPION) and coated (SPION-PLL) complexes were evaluated by a transmission electron microscope and the zeta potential. Results: PLL at 0.75μg/ml showed optimal results with 25 μg/ml SPION concentration compared with other PLL concentrations (0.25, 0.50, 1 and 2 μg/ml). The 18% efficiency with the transfected cells showed green fluorescence. Conclusion: Transfection with SPION is an efficient, non-viral gene transfere method.}, Keywords = {Neural Stem cells (NSC), Nanoparticles, Transfection}, volume = {17}, Number = {2}, pages = {71-76}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1114.2013}, url = {http://ibj.pasteur.ac.ir/article-1-947-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-947-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Afzali, Masoumeh and Nakhaee, Alireza and Tabatabaei, Seyed Payman and Tirgar-Fakheri, Kourosh and Hashemi, Mohamm}, title = {Aberrant Promoter Methylation Profile of Niemann-Pick Type C1 Gene in Cardiovascular Disease}, abstract ={Background: The protein of Niemann-pick type C1 (NPC1) gene promotes the egress of cholesterol from late endosomes and lysosomes to other cellular compartments and contributes to a process known as reverse cholesterol transport. This study aimed to examine whether promoter methylation of NPC1 is associated with risk of cardiovascular disease (CVD). Methods: Fifty CVD patients and 50 healthy subjects as the control group were recruited in this study. Promoter methylation of NPC1 gene was defined using a nested-methylation specific polymerase chain reaction method. Statistical analyses were done using the chi-square, t-test or ANOVA tests. Results: Our study showed that the frequency of semi-methylated promoter (methylated/unmethylated status) was significantly higher in CVD patients than that in controls (OR = 6.521, 95% CI = 2.211-19.215, P = 0.008). However, a completely methylated promoter (methylated/methylated status) was not detected in any subjects in either of the two groups tested. Additionally, the analysis of clinical data according to the methylation status of NPC1 gene demonstrated that serum levels of total cholesterol, total triglycerides, high low-density lipoprotein cholesterol (LDL-C) and low high-density lipoprotein cholesterol (HDL-C) are influenced by NPC1 methylation, so that subjects with a completely unmethylated promoter (Unmethylated/unmethylated status) held lower levels of total triglycerides, total cholesterol, LDL-C and higher levels of HDL-C. Conclusion: Our findings propose that the NPC1 promoter methylation is a probable mechanism that can result in reduced/impaired NPC1 expression/activity and may thus contribute to progression of CVD.}, Keywords = {Niemann-pick type C1 (NPC1), Promoter methylation, cardiovascular disease}, volume = {17}, Number = {2}, pages = {77-83}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.11432.2013}, url = {http://ibj.pasteur.ac.ir/article-1-919-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-919-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {BabaahmadiRezaei, Hossein and Doosti, Mahmood and Aminian, Mahdi and Shabani, Paris}, title = {Compare the Effect of Eicosapentaenoic Acid and Oxidized Low-Density Lipoprotein on the Expression of CD36 and Peroxisome Proliferator-Activated Receptor Gamma}, abstract ={Background: There is evidence that CD36 promotes foam cell formation through internalizing oxidized LDL (ox-LDL) into macrophages therefore, it plays a key role in pathogenesis of atherosclerosis. In addition, CD36 expression seems to be mediated by nuclear receptor peroxisome proliferator-activated receptor gamma (PPAR-γ). The aim of the present study was to evaluate and compare the effect of PPAR-γ ligands, eicosapentaenoic acid (EPA) as an anti-atherogenic factor and ox-LDL as an atherogenic factor on CD36 expression. Mechanism of PPAR-γ action and its ligands in CD36 expression were also investigated. Methods: Raw 264.7 macrophage cell line was treated with ox-LDL (100 and 150 μg protein/LDL) and EPA (100 and 200 μM) for 24 and 48 hours in absence or presence of PPAR-γ inhibitor, T0070907. Quantitative real-time PCR and Western-blotting were used for analysis of gene and protein expression, respectively. Results: Raw 264.7 exposures to ox-LDL and EPA resulted in increased expression of CD36 mRNA and protein however, mRNA and PPAR-γ protein were not up-regulated significantly. Pre-incubation of cells with T0070907 led to decreased expression of CD36 when treated with ox-LDL and EPA. Conclusion: It was confirmed that both EPA and ox-LDL increased CD36 expression but not PPAR-γ, and also co-treatment with PPAR-γ inhibitor decreased CD36 expression. We concluded that up-regulation of CD36 depends on PPAR-γ activation and is not related to increased expression of PPAR-γ. Induction of CD36 by EPA showed that CD36 suppression is not the means by which ω-3 fatty acids (EPA) provide protection against formation of atherosclerotic plaque.}, Keywords = {Atherosclerosis, proliferator-activated receptor gamma (PPAR-γ), Oxidized low density lipoprotein (ox-LDL), Eicosapentaenoic acid (EPA)}, volume = {17}, Number = {2}, pages = {84-92}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.11322.2013}, url = {http://ibj.pasteur.ac.ir/article-1-933-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-933-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Mani, Monireh and Golmohammadi, Taghi and Khaghani, Shahnaz and Zamani, Zahra and Azadmanesh, Kayhan and Meshkani, Reza and Pasalar, Parvi}, title = {Homocysteine Induces Heme Oxygenase-1 Expression via Transcription Factor Nrf2 Activation in HepG2 Cells}, abstract ={Background: Elevated level of plasma homocysteine has been related to various diseases. Patients with hyperhomocysteinemia can develop hepatic steatosis and fibrosis. We hypothesized that oxidative stress induced by homocysteine might play an important role in pathogenesis of liver injury. Also, the cellular response designed to combat oxidative stress is primarily controlled by the transcription factor Nrf2, a principal inducer of anti-oxidant and phase II-related genes. Methods: HepG2 cells were treated with homocysteine in different time periods. Glutathione content was measured by flowcytometry. Using electrophoretic mobility shift assay (EMSA) and Western-blotting, anti-oxidant response element (ARE)-binding activity of Nrf2 for heme ocygenase-1 (HO-1) was demonstrated. Real time RT-PCR and Western-blotting were performed to evaluate whether homocysteine was able to induce mRNA and protein expression of HO-1. Results: The role of Nrf2 in cellular response to homocysteine is substantiated by the following observations in HepG2 cells exposed to homocysteine (i) Western-blotting revealed that Nrf2 is strongly stabilized and became detectable in nuclear extracts. (ii) EMSA demonstrated increased binding of Nrf2 to oligomers containing HO-1 promoter-specific ARE-binding site. (iii) Real time RT-PCR and Western-blotting revealed increased mRNA and protein expression of inducible gene HO-1 after treatment with homocysteine. Conclusion: Data presented in the current study provide direct evidence that the immediate cellular response to oxidative stress provoked by homocysteine is orchestrated mainly by the Nrf2-ARE pathway. Therefore, induction of Nrf2-ARE-dependent expression of HO-1 could be a therapeutic option for hepatic cells damage induced by homocysteine.}, Keywords = {Heme oxygenase-1, HepG2 cells, Oxidative stress}, volume = {17}, Number = {2}, pages = {93-100}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1158.2013}, url = {http://ibj.pasteur.ac.ir/article-1-951-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-951-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Bahrami, Farideh and Janahmadi, Mahyar}, title = {Antibiotic Supplements Affect Electrophysiological Properties and Excitability of Rat Hippocampal Pyramidal Neurons in Primary Culture}, abstract ={Introduction: Antibiotic supplements are regularly used in neuronal culture media to control contamination however, they can interfere with the neuronal excitability and affect electrophysiological properties. Therefore, in this study, the effect of penicillin/streptomycin supplements on the spontaneous electrophysiological activity of hippocampal pyramidal neurons was examined. Methods: Electrophysiological whole-cell patch-clamp recordings from rat hippocampal pyramidal cells in primary culture were performed to investigate the effects of antibiotic supplements on the intrinsic excitability of cultured cells. Results: The present findings indicated that presence of antibiotic supplements (penicillin/streptomycin) in the culture medium altered the intrinsic electrical activity of hippocampal pyramidal neurons in primary culture. These alterations included: 1) depolarized resting membrane potential 2) a significant enhancement in the after-hyperpolarization amplitude 3) a significant increase in the area under the action potential and in the decay and rise time of the action potential 4) a significant broadening of action potential and 5) a significant reduction in the firing frequency. Conclusion: These findings suggest that addition of antibiotic supplements to culture media influences the neuronal excitability and alters the electrophysiological properties of cultured neurons, possibly through changing the ionic conductance underlying neuronal excitability.}, Keywords = {Primary cell culture, Patch-clamp techniques, Hippocampus}, volume = {17}, Number = {2}, pages = {101-106}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.11242.2013}, url = {http://ibj.pasteur.ac.ir/article-1-903-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-903-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {CarvalhodeM.Thiele, Magna and CarlosBohn, Carlos Bohn and LimaChaiben, Cassiano and TrindadeGrégio, Ana Maria and ÂngelaNavalMachado, Maria and AdilsonSoaresdeLima, Antonio}, title = {Nucleolar Organizer Regions of Oral Epithelial Cells in Crack Cocaine Users}, abstract ={Background: The health risks of crack cocaine smoking on the oral mucosa has not been widely researched and documented. Objective: The purpose of this study was to analyze the proliferative activity of oral epithelial cells exposed to crack cocaine smoke using silver nucleolar organizer region (AgNOR) staining. Methods: Oral smears were collected from clinically normal-appearing buccal mucosa by liquid-based exfoliative cytology of 60 individuals (30 crack cocaine users and 30 healthy controls matched for age and gender) and analyzed for cytomorphologic and cytomorphometric techniques. Results: Crack cocaine users consumed about 13.3 heat-stable rocks per day and the time consumption of the drug was of 5.2 (± 3.3) years. Mean values of AgNOR counting for case and control groups were 5.18 ± 1.83 and 3.38 ± 1.02 (P<0.05), respectively. AgNOR area and percentage of AgNOR-occupied nuclear area were increased in comparison with the control (P<0.05). There was no statistically significant difference in the mean values of the nuclear area between the groups (P>0.05). Conclusion: This study revealed that crack cocaine smoke increases the rate of cellular proliferation in cells of normal buccal mucosa.}, Keywords = {Crack-Cocaine, Mouth mucosa, Cell proliferation}, volume = {17}, Number = {2}, pages = {107-111}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.11152.2013}, url = {http://ibj.pasteur.ac.ir/article-1-934-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-934-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Zaminy, Arash and Shokrgozar, Mohammad Ali and Sadeghi, Yousef and Noroozian, Mohsen and Heidari, Mohammad Hassan and Piryaei, Abbas}, title = {Mesenchymal Stem Cells as an Alternative for Schwann Cells in Rat Spinal Cord Injury}, abstract ={Background: Spinal cord has a limited capacity to repair therefore, medical interventions are necessary for treatment of injuries. Transplantation of Schwann cells has shown a great promising result for spinal cord injury (SCI). However, harvesting Schwann cell has been limited due to donor morbidity and limited expansion capacity. Furthermore, accessible sources such as bone marrow stem cells have drawn attentions to themselves. Therefore, this study was designed to evaluate the effect of bone marrow-derived Schwann cell on functional recovery in adult rats after injury. Methods: Mesenchymal stem cells were cultured from adult rats’ bone marrow and induced into Schwann cells in vitro. Differentiation was confirmed by immunocytochemistry and RT-PCR. Next, Schwann cells were seeded into collagen scaffolds and engrafted in 3 mm lateral hemisection defects. For 8 weeks, motor and sensory improvements were assessed by open field locomotor scale, narrow beam, and tail flick tests. Afterwards, lesioned spinal cord was evaluated by conventional histology and immunohistochemistry. Results: In vitro observations showed that differentiated cells had Schwann cell morphology and markers. In this study, we had four groups (n = 10 each): laminectomy, control, scaffold and scaffold + Schwann cells. Locomotor and sensory scores of cell grafted group were significantly better than control and scaffold groups. In histology, axonal regeneration and remyelination were better than control and scaffold groups. Conclusion: This study demonstrates that bone marrow-derived Schwann cells can be considered as a cell source for Schwann cells in SCI treatment.}, Keywords = {Rats, Spinal cord injuries (SCI), Bone marrow, Schwann cells, Cell transdifferentiation}, volume = {17}, Number = {3}, pages = {113-122}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1121.2013}, url = {http://ibj.pasteur.ac.ir/article-1-953-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-953-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Amoushahi, Mahboobeh and Salehnia, Mojdeh and HosseinKhani, Sam}, title = {The Effect of Vitrification and in vitro Culture on the Adenosine Triphosphate Content and Mitochondrial Distribution of Mouse Pre-Implantation Embryos}, abstract ={Background: The mitochondria are an important source of adenosine triphosphate (ATP) production in pre-implantation embryo. Therefore, the objective of this study was to investigate the effect of vitrification and in vitro culture of mouse embryos on their mitochondrial distribution and ATP content. Methods: The embryos at 2-PN, 4-cell and blastocyst stages were collected from the oviduct of stimulated pregnant mice and uterine horns. Then, the embryos were vitrified with the cryotop method using ethylene glycol and dimethylsulphoxide. After evaluating the survival rates of vitrified embryos, their development to hatching stages were assessed. The ATP content of collected in vivo and in vitro embryos at different stages was measured by luciferin-luciferase bioluminescence assay. The distribution of mitochondria was studied using Mito-tracker green staining under a fluorescent microscope. Results: The survival rates of vitrified embryos at 2-PN, 4-cell and early blastocyst stages were 84.3, 87.87 and 89.89%, respectively. The hatching rates in previous developmental stages in vitrified group were 57.44, 66.73 and 70.89% and in non-vitrified group were 66.32, 73.25 and 75.89%, respectively (P>0.05) The ATP content of in vivo or in vitro collected embryos was not significantly different in both vitrified and non-vitrified groups (P>0.05). Mitochondrial distribution of vitrified and non-vitrified 2-PN embryos was similar, but some clampings or large aggregation of mitochondria within the vitrified 4-cell embryos was prominent. Conclusions: Vitrification method did not affect the mouse embryo ATP content. Also, the cellular stress was not induced by this procedure and the safety of vitrification was shown.}, Keywords = {Mitochondria, Vitrification, Adenosine triphosphate (ATP)}, volume = {17}, Number = {3}, pages = {123-128}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1199.2013 }, url = {http://ibj.pasteur.ac.ir/article-1-967-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-967-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Doosti, Masoumeh and Ramazani, Ali and Garshasbi, Maryam}, title = {Identification and Characterization of Metallo-β-Lactamases Producing Pseudomonas aeruginosa Clinical Isolates in University Hospital from Zanjan Province, Iran}, abstract ={Background: Infectious by Pseudomonas aeruginosa has spread worldwide and metallo-beta-lactamases (MBL) are being reported with increasing frequency. The aim of this study was to investigate the antibiotic susceptibility and distribution of blaVIM and blaIMP genes in P. aeruginosa isolates from Zanjan Province of Iran. Methods: A total of 70 P. aeruginosa isolates were identified from patients admitted at intensive care units. The antimicrobial susceptibility was tested by disk diffusion (Kirby-Bauer) method and for production of MBL using double-disk synergy test (DDST). After DNA extraction, the presence of blaVIM and blaIMP genes and class 1 integron were detected by PCR. RESULTS: Most of the isolates were resistant to meropenem, cefotaxime and imipenem (IPM). Also, 44/70 (62.85%) IPM resistant isolates were confirmed by DDST. Of the 44 clinical isolates, 41 (93%) isolates showed MIC≥4 µg/ml for IPM. Based on the DDST results, 36 (87.8%) were confirmed to be MBL producers. PCR amplification showed that 23/41 (56%) carried blaVIM and 10/41 (24.3%) possessed blaIMP gene. Also, 31/44 (70.5%) isolates contained class 1 integron gene. Conclusion: Our results highlight that the genes for Verona integron-encoded metallo-β-lactamase, IPM β-lactamases and class 1 integrons were predominantly present among the IPM-resistant P. aeruginosa tested in our province and also the frequency of blaVIM type is higher than blaIMP. This is the first report of P. aeruginosa strains producing blaIMP with high frequency from Zanjan province of Iran.}, Keywords = {Pseudomonas aeruginosa, Beta-lactamases, PCR}, volume = {17}, Number = {3}, pages = {129-133}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1107.2013}, url = {http://ibj.pasteur.ac.ir/article-1-960-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-960-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Daniali, Samira and Nahavandi, Arezo and Madjd, Zahra and Shahbazi, Ali and Niknazar, Somayeh and Shahbazzadeh, Delavar}, title = {Chronic Ritalin Administration during Adulthood Increases Serotonin Pool in Rat Medial Frontal Cortex}, abstract ={Background: Ritalin has high tendency to be abused. It has been the main indication to control attention deficit hyperactivity disorder. The college students may seek for it to improve their memory, decrease the need for sleep (especially during exams), which at least partially, can be related to serotonergic system. Therefore, it seems worthy to evaluate the effect of Ritalin intake on mature brain. There are many studies on Ritalin effect on developing brain, but only few studies on adults are available. This study was undertaken to find Ritalin effect on serotonin transporter (SERT) density in medial frontal cortex (MFC) of mature rat. Methods: Thirty male Wistar rats were used in the study. Rats were assigned into five groups (n = 6 per group): one control, two Ritalin and two vehicle groups. Twelve rats received Ritalin (20 mg/kg/twice a day) orally for eleven continuous days. After one week of withdrawal and another two weeks of rest, in order to evaluate short-term effects of Ritalin, six rats were sacrificed. Another six rats were studied to detect the long-term effects of Ritalin therefore, they were sacrificed 12 weeks after the previous group. The immunohistochemistry was performed to evaluate the results. Results: Immunohistochemistry studies showed a higher density of SERT in both 2 and 12 weeks after withdrawal from Ritalin intake in MFC of rat and there was no significant difference between these two groups. Conclusions: Our findings demonstrated both short- and long-term effects of Ritalin on frontal serotonergic system after withdrawal period.}, Keywords = {Ritalin, Serotonin, Rats}, volume = {17}, Number = {3}, pages = {134-139}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1173.2013 }, url = {http://ibj.pasteur.ac.ir/article-1-980-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-980-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Edalat, Houri and Hajebrahimi, Zahra and Pirhajati, Vahid and Movahedin, Mansoureh and Tavallaei, Mahmoud and Soroush, Mohammad-Reza and Mowla, Seyed Jav}, title = {Transplanting P75-Suppressed Bone Marrow Stromal Cells Promotes Functional Behavior in a Rat Model of Spinal Cord Injury}, abstract ={Background: Bone marrow stromal cells (BMSC) have been successfully employed for movement deficit recovery in spinal cord injury (SCI) rat models. One of the unsettled problems in cell transplantation is the relative high proportion of cell death, specifically after neural differentiation. According to our previous studies, p75 receptor, known as the death receptor, is only expressed in BMSC in a time window of 6-12 hours following neural induction. Moreover, we have recently reported a decreased level of apoptosis in p75-suppressed BMSC in vitro. Therefore, our objective in this research was to explore the functional effects of transplanting p75:siRNA expressing BMSC in SCI rats. Methods: Laminectomy was performed at L1 vertebra level to expose spinal cord for contusion using weight-drop method. PBS-treated SCI rats (group one) were used as negative controls, in which cavitations were observed 10 weeks after SCI. pRNA-U6.1/Hygro- (group two, as a mock) and pRNA-U6.1/Hygro-p75 shRNA- (group three) transfected BMSC were labeled with a fluorescent dye, CM-DiI, and grafted into the lesion site 7 days after surgery. The Basso-Beattie-Bresnehan locomotor rating scale was performed weekly for 10 weeks. Results: There was a significant difference (P≤0.05) between all groups of treated rats regarding functional recovery. Specifically, the discrepancy among p75 siRNA and mock-transfected BMSC was statistically significant. P75 siRNA BMSC also revealed a higher level of in vivo survival compared to the mock BMSC. Conclusion: Our data suggest that genetically modified BMSC that express p75:siRNA could be a more suitable source of cells for treatment of SCI.}, Keywords = {Spinal cord injury (SCI), Apoptosis, Bone marrow stromal cells (BMSC)}, volume = {17}, Number = {3}, pages = {140-145}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1193.2013}, url = {http://ibj.pasteur.ac.ir/article-1-974-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-974-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Attar, Armin and KhosraviMaharlooi, Mohsen and Khoshkhou, Sara and Hosseini, Ahmad and Jaberipour, Mansoureh and Dehghan, Arman and Monabati, Ahm}, title = {Colony Forming Unit Endothelial Cells Do not Exhibit Telomerase Alternative Splicing Variants and Activity}, abstract ={Introduction: Endothelial progenitor colony forming unit-endothelial cells (CFU-EC) were first believed to be the progenitors of endothelial cells, named endothelial progenitor cells. Further studies revealed that they are monocytes regulating vasculogenesis. The main hindrance of these cells for therapeutic purposes is their low frequency and limited replicative potentials. This study was undertaken to determine telomerase activity and alternative splicing variants in CFU-EC as a potential cause of limited replicative capacity in these cells. Methods: CFU-EC were isolated from peripheral blood using a standard cell culture assay. Colonies were detached mechanically and alternative splicing variant mRNA were evaluated using real-time PCR. Telomerase enzyme activity was assessed using telomerase repeat amplification protocol. The same procedures were done on the cancer cell line Calu6 as the positive control. Results: The cultured peripheral blood mononuclear cells formed colonies with spindle-shaped monocytic cells sprouted from the clusters. These morphological characteristics fulfill the definition of CFU-EC. Telomere length amplification protocol assay revealed no telomerase activity and real-time PCR showed no expression of telomerase enzyme mRNA in CFU-EC. Both parameters were significantly higher in the cancer cell line Calu6 taken as the positive control. Conclusion: The absence of telomerase activity in the CFU-EC is a result of pre-transcriptional regulation of gene expression rather than other mechanisms for controlling telomerase activity such as post-transcriptional modifications. This finding can explain the limited proliferative activity of CFU-EC cells. We propose that absence of telomerase activity in CFU-EC can be attributable to their more mature monocytic nature that needs further investigations.}, Keywords = {Telomerase, Endothelial cells, Hemangioblast, Alternative splicing}, volume = {17}, Number = {3}, pages = {146-151}, publisher = {Pasteur Institute of Iran}, doi = { 10.6091/ibj.1100.2013}, url = {http://ibj.pasteur.ac.ir/article-1-957-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-957-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Mohammadi, Mohammad Taghi and Amini, Reza and Jahanbakhsh, Zahra and Shekarforoush, Shahnaz}, title = {Effects of Atorvastatin on the Hypertension-Induced Oxidative Stress in the Rat Brain}, abstract ={Background: It is well known that the development of brain oxidative stress is one of the most serious complications of arterial hypertension that evokes brain tissue damage. The aim of this study was to examine the effects of atorvastatin treatment (20 mg/kg/day), as an antioxidant, to prevent the brain tissue oxidative stress in the hypertensive (HTN) rats. Methods: Experiments were performed in four groups of rats (n = 5 each group): sham, sham-treated, HTN and HTN treated. Rats were made HTN by aortic constriction above the renal arteries. After 30 days, rats were slaughtered under deep anesthesia to remove brain hemispheres. After tissue homogenization, enzyme activities of superoxide dismutase (SOD) and catalase (CAT), as well as glutathione (GSH) content and malondialdehyde (MDA) level were determined by biochemical methods. Results: In HTN rats, arterial blood pressure was increased about 40% and brain enzyme activities of SOD and CAT were significantly decreased compared with sham group. Induction of hypertension significantly decreased GSH content and increased MDA level of brain tissue. Treatment with atorvastatin enhanced the activity of SOD and prevented from GSH decrement during hypertension. Conclusion: Based on the findings of this study, treatment with atorvastatin might have saved the brain tissue of HTN rats from hypertension-induced oxidative stress.}, Keywords = {Atorvastatin, Aortic coarctation, Oxidative stress, Hypertension}, volume = {17}, Number = {3}, pages = {152-157}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1189.2013 }, url = {http://ibj.pasteur.ac.ir/article-1-964-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-964-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {SharifiTabar, Mehdi and Habashi, Ali Akbar and RajabiMemari, Hami}, title = {Human Granulocyte Colony-Stimulating Factor (hG-CSF) Expression in Plastids of Lactuca sativa}, abstract ={Background: Human granulocyte colony-stimulating factor (hG-CSF) can serve as valuable biopharmaceutical for research and treatment of the human blood cancer. Transplastomic plants have been emerged as a new and high potential candidate for production of recombinant biopharmaceutical proteins in comparison with transgenic plants due to extremely high level expression, biosafety and many other advantages. Methods: hG-CSF gene was cloned into pCL vector between prrn16S promoter and TpsbA terminator. The recombinant vector was coated on nanogold particles and transformed to lettuce chloroplasts through biolistic method. Callogenesis and regeneration of cotyledonary explants were obtained by Murashige and Skoog media containing 6-benzylaminopurine and 1-naphthaleneacetic acid hormones. The presence of hG-CSF gene in plastome was studied with four specific PCR primers and expression by Western immunoblotting. Results: hG-CSF gene cloning was confirmed by digestion and sequencing. Transplastomic lettuce lines were regenerated and subjected to molecular analysis. The presence of hG-CSF in plastome was confirmed by PCR using specific primers designed from the plastid genome. Western immunoblotting of extracted protein from transplastomic plants showed a 20-kDa band, which verified the expression of recombinant protein in lettuce chloroplasts. Conclusions: This study is the first report that successfully express hG-CSF gene in lettuce chloroplast. The lettuce plastome can provide a cheap and safe expression platform for producing valuable biopharmaceuticals for research and treatment.}, Keywords = {Plastid, Human granulocyte colony-stimulating factor (hG-CSF), Biolistics, Gene targeting}, volume = {17}, Number = {3}, pages = {158-164}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1180.2013}, url = {http://ibj.pasteur.ac.ir/article-1-968-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-968-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Sayadmanesh, Ali and Ebrahimi, Firouz and Hajizade, Abbas and Rostamian, Mosayeb and Keshavarz, Hani}, title = {Expression and Purification of Neurotoxin-Associated Protein HA-33/A from Clostridium botulinum and Evaluation of Its Antigenicity}, abstract ={Background: Botulinum neurotoxin (BoNT) complexes consist of neurotoxin and neurotoxin-associated proteins. Hemagglutinin-33 (HA-33) is a member of BoNT type A (BoNT/A) complex. Considering the protective role of HA-33 in preservation of BoNT/A in gastrointestinal harsh conditions and also its adjuvant role, recombinant production of this protein is favorable. Thus in this study, HA-33 was expressed and purified, and subsequently its antigenicity in mice was studied. Methods: Initially, ha-33 gene sequence of Clostridium botulinum serotype A was adopted from GenBank. The gene sequence was optimized and synthesized in pET28a (+) vector. E. coli BL21 (DE3) strain was transformed by the recombinant vector and the expression of HA-33 was optimized at 37°C and 5 h induction time. Results: The recombinant protein was purified by nickel nitrilotriacetic acid agarose affinity chromatography and confirmed by immunoblotting. Enzyme Linked Immunoassay showed a high titer antibody production in mice. Conclusion: The results indicated a highly expressed and purified recombinant protein, which is able to evoke high antibody titers in mice.}, Keywords = {Botulinum neurotoxin, Expression, Purification}, volume = {17}, Number = {4}, pages = {165-170}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1216.2013}, url = {http://ibj.pasteur.ac.ir/article-1-1023-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-1023-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Habibi, Laleh and Shokrgozar, Mohammad Ali and Motamedi, Mahdieh and Akrami, Seyed Mohamm}, title = {Effect of Heavy Metals on Silencing of Engineered Long Interspersed Element-1 Retrotransposon in Nondividing Neuroblastoma Cell Line}, abstract ={Background: L1 retrotransposons are the most active mobile DNA elements in human genome. Unregulated L1 retrotransposition may have deleterious effect by disrupting vital genes and inducing genomic instabilities. Therefore, human cells control L1 elements by silencing their activities through epigenetic mechanisms. It has been shown that cell division and heavy metals stimulate the frequency of L1 activities. Removal of silencing by L1 motivators may restart L1 element functions. Here, we have proposed that weather neurotoxic environmental heavy metals (as L1 stimulating factors) have a role in removing L1 silencing and restating its activities in nondividing neuronal cells. Methods: L1-RP green fluorescent protein (GFP)-tagged knock-in human neuroblastoma clones were prepared. Single-cell clone was treated with mitomycin-c combined with nontoxic and toxic concentrations of iron (Fe), copper (Cu), and mercury (Hg). Silencing status of engineered L1 elements in dividing and nondividing cells was determined through measuring the amount of GFP expressing cells with flow cytometry. The cytotoxic effect of mitomycin-c combined with metals was measured by MTT assay. Results: Hg in nondividing cells and Fe, Cu, and Hg in dividing neuroblastoma cells could significantly remove L1 silencing. Also, mitomycin-c treatment did not have any effect on metal toxicity status in neuroblastoma cells. Conclusion: Totally, our findings have shown that cell division has a role in removing L1 silencing as well as L1 retrotransposition induced by environmental heavy metals. It has been also indicated that Hg at all concentrations could remove silencing of engineered L1 element regardless of cell cycle state.}, Keywords = {Cell division, Heavy metals, L1 retrotransposon}, volume = {17}, Number = {4}, pages = {171-178}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1230.2013}, url = {http://ibj.pasteur.ac.ir/article-1-1022-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-1022-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Abdollahi, Maasoume and Salehnia, Mojdeh and Salehpour, Saghar and Ghorbanmehr, Nassim}, title = {Human Ovarian Tissue Vitrification/Warming Has Minor Effect on the Expression of Apoptosis-Related Genes}, abstract ={Background: In this study, we evaluated the incidence of apoptosis at the ultrastructural levels and expression of some apoptosis-related genes in vitrified human ovarian tissue just after warming. Methods: Human ovarian tissue biopsies from 23 women after caesarean section were transported to the laboratory within 2 hours, and then they were cut into small pieces. Some pieces were vitrified and warmed and the other samples were considered as control. Apoptosis was assessed by a transmission electron microscope and also by molecular analysis of pro-apoptotic (Fas, FasL, Bax, p53, caspase8, and caspase3) and antiapoptotic (Bcl-2 and BIRC5) genem RNA levels using real-time RT-PCR before and after vitrification. Results: No sign of apoptosis was shown ultrastructurally in vitrified samples. The level of FasL, Bcl-2, Bax, p53, and caspase3 mRNA and Bax:Bcl-2 ratio were similar in non-vitrified and vitrified groups however, the expression of Fas and caspase8 genes was higher and BIRC5 was lower in vitrified samples compared to non-vitrified group (P<0.05). Conclusion: The fine structure of human vitrified ovarian tissue was well preserved moreover, vitrification was shown to affect the expression of some apoptosis-related genes. However, additional study is needed to confirm this observation.}, Keywords = {Vitrification, Apoptosis, Gene expression, Ovary, Humans}, volume = {17}, Number = {4}, pages = {179-186}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1243.2013}, url = {http://ibj.pasteur.ac.ir/article-1-1011-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-1011-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Yadegari, Maryam and Orazizadeh, Mahmoud and Hashemitabar, Mahmoud and Khodadadi, Ali}, title = {Protective Effects of Interleukin-4 on Tissue Destruction and Morphological Changes of Bovine Nasal Chondrocytes in vitro}, abstract ={Background: Previous studies have shown that some cytokines have protective effects on cartilage in joint diseases. In the current study, effects of IL-4 against morphological changes and tissue degradation induced by IL-1α on bovine nasal cartilage (BNC) explants were investigated. Methods: Fresh BNC samples were prepared from a slaughterhouse under sterile conditions. BNC explants culture was treated with both IL-lα (10 ng/ml) and IL-4 (50 ng/ml) at the same time for 28 days. The morphological characteristics of explants were assessed by using histology techniques and invert microscopy. Matrix metalloproteinase-1 (MMP-1) production was assessed within different days by using Western blotting. Results: IL-lα induced prominent cartilage morphology degradation. The pro and active form of MMP-1 band substantially increased at day 21 of culture. In the presence of both IL-lα and IL-4, chondrocytes preserved their ordinary normal phenotype with intact extracellular matrix. In addition, a significant reduction in pro-MMP-1and inhibition of active MMP-1 was seen. Conclusion: In conclusion, IL-4 could be regarded as a potential candidate in cartilage protecting against the degradation changes of IL-lα. It seems that the preservation effect of IL-4 is associated with significant reduction of MMP-1.}, Keywords = {Chondrocyte, Interleukin-1α, Interleukin-4, Matrix metalloproteinase-1, Bovine nasal cartilage}, volume = {17}, Number = {4}, pages = {187-193}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1219.2013}, url = {http://ibj.pasteur.ac.ir/article-1-1032-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-1032-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Hashemi, Mohammad and Zakeri, Zahra and Eskandari-Nasab, Ebrahim and Atabaki, Mahdi and Pourhosseini, Seyed Mohammad Ebrahim and Jahantigh, Mehdi and Bahari, Gholamreza and Taheri, Mohse}, title = {CD226 rs763361 (Gly307Ser) Polymorphism Is Associated with Susceptibility to Rheumatoid Arthritis in Zahedan, Southeast Iran}, abstract ={Background: Rheumatoid arthritis (RA) is a chronic inflammatory disease with many genetic factors predisposing to disease susceptibility. The aim of the present study was to investigate the impact of CD226 rs727088 and rs763361 polymorphisms and susceptibility to RA in a sample of the Iranian population. Methods: This case-control study was carried out on 100 patients with RA and 104 healthy subjects. The polymorphisms were determined using tetra amplification refractory mutation system-polymerase chain reaction assay. Results: The rs763361 (Gly307Ser) polymorphism increased the risk of RA in codominant, dominant and recessive-tested inheritance models (odds ratio [OR] = 3.18, 95% confidence intervals [95% CI] = 1.44-7.02, P = 0.004, CC vs. TT, and OR = 1.98, 95% CI = 1.10-3.57, P = 0.023, CC vs. CT-TT, and OR = 2.61, 95% CI = 1.26-5.37, P = 0.010, CC + CT vs. TT, respectively). In addition, the rs763361 T allele increased the risk of RA (OR = 2.06, 95% CI = 1.38-3.08, P<0.001). However, no significant difference was observed among the groups regarding CD226 rs727088 polymorphism (χ2 = 3.20, P = 0.202). Conclusions: Our finding showed that CD226 rs763361, but not rs727088, gene polymorphism increased the risk of RA in a sample of the Iranian population.}, Keywords = {Rheumatoid arthritis (RA), CD226, Polymorphism}, volume = {17}, Number = {4}, pages = {194-199}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1205.2013}, url = {http://ibj.pasteur.ac.ir/article-1-1009-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-1009-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Rajaei, Farzad and Abedpour, Neda and Salehnia, Mojdeh and Jahanihashemi, Hass}, title = {The Effect of Vitrification on Mouse Oocytes Apoptosis by Cryotop Method}, abstract ={Background: Oocyte cryopreservation is one of the most important topics in the field of assisted reproductive technology to preserve women fertility, but relationship between cryopreservation and apoptosis is still a matter of debate. The present study was aimed to investigate the effects of vitrification on apoptosis in mouse oocytes by Cryotop method. Method: A total of 200 germinal vesicle (GV) and 200 metaphase II (MII) oocytes were obtained from ovaries and fallopian tubes of NMRI mice, respectively and divided into control and experimental groups. Oocytes in experimental group were vitrified by Cryotop using vitrification medium and were kept in liquid nitrogen for one month. The survival rate of oocytes was evaluated after 2 hour incubation time. Then, the oocyte apoptosis was evaluated by TUNEL technique and compared with those in control group. The data was compared statistically using SPSS software and chi-square test. Results: The survival rates of vitrified GV (93%) and MII oocytes (88%) showed a significant decrease compared with the control group (P<0.05), but there was no significant difference in survival rate of both vitrified oocyte groups. The incidence of apoptosis in vitrified and control GV oocytes showed no significant difference (13% vs. 7%), but the rate of apoptosis in vitrified MII oocytes increased significantly not only in comparison with MII control group (25% vs. 5%) but also with vitrified GV oocytes (P<0.05). Conclusion: The results indicate that vitrification increases apoptosis in mouse MII oocytes and apoptosis may play a role in MII oocyte injury after vitrification.}, Keywords = {Vitrification, Apoptosis, Oocytes}, volume = {17}, Number = {4}, pages = {200-205}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1184.2013}, url = {http://ibj.pasteur.ac.ir/article-1-1010-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-1010-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Khordad, Elnaz and Fazel, Alireza and EbrahimzadehBideskan, Alirez}, title = {The Effect of Ascorbic Acid and Garlic Administration on Lead-Induced Apoptosis in Rat Offspring\'s Eye Retina}, abstract ={Introduction: Lead toxicity induces retinal cell apoptosis. Vitamin C and garlic may decrease lead-induced apoptosis. This study was undertaken to investigate vitamin C and garlic protective effects on lead-induced apoptosis in eye retina. Methods: Pregnant Wistar rats (n = 72) were divided randomly into 9 groups: (L) treated rats with lead acetate in drinking water and (L+AA) with leaded water and vitamin C intraperitoneally(L+G), the rats received leaded-water and garlic juice via gavage (L+AA+G) treated rats with leaded water, ascorbic acid, and garlic juice, (AA) with ascorbic acid, and (G) with garlic juice (AA+G) treated rats with vitamin C and garlic juice and (Sh) with tap water plus normal hydrogen chloride (HCl) and glucose normal (N). After 21-day lactation, blood lead level (BLL) in rats was measured, and then their offspring and the rat offspring's eyes were removed and processed for using TUNEL method. TUNEL positive cells in the eye retina were counted and all groups were compared. Results: BLL increased in L group compared to the control groups and decreased significantly in L + G, L + AA, and L+ AA + G groups compared to L group (P<0.05). TUNELL positive cell number in eye retina significantly increased in L group compared to control groups (P<0.05) and decreased in L+ G, L+ AA, and L+AA + G groups compared to L group (P<0.05). Conclusion: Garlic juice and ascorbic acid administration during pregnancy and lactation may protect lead-induced apoptosis in rat offspring's eye retina.}, Keywords = {Lead, Garlic, Ascorbic acid, Apoptosis, Retina}, volume = {17}, Number = {4}, pages = {206-213}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1229.2013}, url = {http://ibj.pasteur.ac.ir/article-1-1021-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-1021-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Abdanipour, Alireza and Tiraihi, Taki and Taheri, Taher and Kazemi, Hadi}, title = {Microglial Activation in Rat Experimental Spinal Cord Injury Model}, abstract ={Background: The present study was designed to evaluate the secondary microglial activation processes after spinal cord injury (SCI). Methods: A quantitative histological study was performed to determine ED-1 positive cells, glial cell density, and cavitation size in untreated SCI rats at days 1, 2, and 4, and weeks 1, 2, 3, and 4. Results: The results of glial cell quantification along the 4900-µm long injured spinal cord showed a significant increase in glial cell density percentage at day 2 as compared to other days. Whereas the highest increase in ED-1 immunoreactive cells (monocyte/phagocyte marker in rats) was observed at day 2 (23.15%) post-injury. Evaluation of cavity percentage showed a significant difference between weeks 3 and 4 post-injury groups. Conclusions: This study provides a new insight into the multiphase immune response to SCI, including cellular inflammation, macrophages/microglia activation, glial cell density, and cavitation. Better understanding of the inflammatory processes associated with acute SCI would permit the development of better therapeutic strategies.}, Keywords = {Spinal cord injuries, Inflammation, Microglia, Macrophages}, volume = {17}, Number = {4}, pages = {214-220}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1213.2013}, url = {http://ibj.pasteur.ac.ir/article-1-1025-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-1025-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} } @article{ author = {Zarei, Fatemeh and Yousofvand, Namdar and Khazaei, Mozafar and Ghanbari, Ali}, title = {Effect of Exogenous Testosterone, Finasteride, and Castration on serum level of Thyroxin}, abstract ={Background: The secretion of thyroxin (T4) as the main hormone of thyroid gland is regulated by androgens. The present study aimed to evaluate the effect of testosterone and finasteride administration and castration on serum levels of T4 and to show the effect of this regulation on total body weight, weight of testis, and the weight of prostate. Methods: Male adult rats (n = 32) were divided into 4 groups (n = 8): Group 1 (control), Group 2 (castration), Group 3 (finasteride: 20 mg/kg/day) and Group 4 (testosterone: 5 mg/kg/day). At the end of the study (35 days), serum level of thyroxin, body weight, weight of testis, and prostate were determined. Results: The data showed that the body weight increased in castrated (P = 0.04) and decreased in testosterone (P = 0.00) groups but did not differ in finasteride (P>0.05) group. There were not any differences in the weight of testis among control, finasteride, and testosterone groups but the weight of prostate increased in testosterone group (P = 0.00) and decreased in castrated (P = 0.03) and finasteride groups (P = 0.04). In addition, the serum level of T4 (nmo/ml) decreased in the three groups: finasteride (P = 0.03), testosterone (P = 0.04), and castrated (P = 0.00). Conclusion: Testosterone in both high and low levels decreased the amount of T4 with a time-dependent manner.}, Keywords = {Finasteride, Rats, Testosterone, Thyroxin}, volume = {17}, Number = {4}, pages = {221-224}, publisher = {Pasteur Institute of Iran}, doi = {10.6091/ibj.1234.2013}, url = {http://ibj.pasteur.ac.ir/article-1-1018-en.html}, eprint = {http://ibj.pasteur.ac.ir/article-1-1018-en.pdf}, journal = {Iranian Biomedical Journal}, issn = {1028-852X}, eissn = {2008-823X}, year = {2013} }