OTHERS_CITABLE DNA Barcoding: a new tool with wide array of applications DNA barcoding is a new term introduced in to scientific literatures by Hebert and coworkers almost a decade ago. The concept of barcoding alone is well-known to the public: a series of black bars printed on many commercial products (Universal Product Code), which are used to distinguish different products. Advances made in molecular biology and molecular techniques late 20th century e.g. sequencing technologies, has inspired scientists to apply barcoding concept to all domains of life by using the unique nature of DNA for each single species, in order to generate a comprehensive library of living organisms on the planet earth. Such an ambitious initiative would result in a global DNA barcode database which will be valuable for biological scientists, medical, governmental and legal agencies as a mean of identification. The first initiative for DNA barcoding was funded in Canada and later on several DNA barcoding campaigns came in to the scene. http://rmm.mazums.ac.ir/article-1-52-en.pdf 2013-10-22 1 2 10.18869/acadpub.rmm.1.2.1 DNA barcoding organisms molecular biology oxidase Mahdi Arzanlou arzanlou@hotmail.com 1 Mycology and Plant Pathology, Plant Protection Department, Faculty of Agriculture, University of Tabriz AUTHOR
OTHERS_CITABLE Molecular approaches to diagnosis of invasive aspergillosis what we know and what we do not know. Invasive aspergillosis (IA) are a major complication in immunocompromized patients where can be serious and rapidly fatal. Early diagnosis and early appropriate antifungal treatment is important in reducing mortality and morbidity. But despite many efforts to develop detection methods, the diagnosis of IA still remains challenging and current conventional methods are limited for adequate diagnosis. New rapid methods which can detect IA early in the course of disease with high sensitivity and specificity are needed since treating these infections at an early stage. Using molecular methods for Aspergillus species identification can be a cost-effective, rapid, discriminatory, and objective approach for delineating Aspergillus species in a clinical microbiology laboratory. PCR techniques for the diagnosis of IA have been studied for more than a decade and are still considered investigational, however until now PCR is not included in current EORTC/MSG diagnostic criteria. http://rmm.mazums.ac.ir/article-1-48-en.pdf 2013-10-22 3 9 10.18869/acadpub.rmm.1.2.3 Invasive aspergillosis PCR Molecular approaches real-time PCR Maryam Moazeni m.nabili@gmail.com 1 Department of Medical Mycology and Parasitology, School of Medicine/Molecular and Cell Biology Research Centre, Mazandaran University of Medical Sciences, Sari, Iran AUTHOR Mojtaba Nabili sadegh_7392008@yahoo.com 2 Department of Medical Mycology and Parasitology, School of Medicine/Molecular and Cell Biology Research Centre, Mazandaran University of Medical Sciences, Sari, Iran AUTHOR Sadegh Khodaveisy badalii@yahoo.com 3 Department of Medical Parasitology and Mycology, Kurdistan University of Medical Sciences, Sanandaj, Iran.Department of Medical Parasitology and Mycolog Tehran University of Medical Sciences, Tehran, Irany, AUTHOR Hamid Badali 4 Department of Medical Mycology and Parasitology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. Molecular and Cell Biology Research Centre (MCBRC), Mazandaran University of Medical Sciences, Sari, Iran. AUTHOR
OTHERS_CITABLE Urinary neutrophil-gelatinase associated lipocalin is a more prognostic biomarker to distinguish antenatal hydronephrosis in neonates Background: Routine diagnostic methods of Vesicoureteral reflux (VUR) are invasive and can cause exposure to radiation and may increase risk of urinary tract infections. Therefore, introducing reliable, non invasive methods might be more interested in pediatric nephrology. The objective of this perspective case control study was to evaluate the prognostic value of urinary neutrophil-gelatinase associated lipocalin (uNGAL) on antenatal hydronephrosis (AH) with and without VUR. Materials and Methods: A total of 50 patients diagnosed with AH 78% males with mean age 5.71± 2.1 years, including 27 AH with VUR and 23 AH without VUR, and 19 normal healthy children 78.9% males with mean age 5.63 ± 1.89 years, were enrolled in this study. Urinary NGAL levels were measured by enzyme linked immunosorbent assay (ELISA). Results: There was a significant difference in uNGAL concentration between AH patients and controls (0.80 ± 0.26 and 0.29 ± 0.27 ng/ml, p<0.0001). However, the levels of uNGAL was not significantly deviated between AH patients with VUR compared to those without VUR (0.84 ± 0.34 vs. 0.75 ± 0.13, p=0.419). Standardization of NGAL based on urinary creatinine (uNGAL/uCr) showed a significantly difference between AH neonates with VUR compared to those without VUR (2.43±1.61 vs. 1.91±0.79, p<0.0001). Receiver operator characteristic (ROC) analysis revealed higher prognostic power of uNGAL for identifying AH with a sensitivity 95.7%, and specificity 84.2%. Meanwhile, the levels of uNGAL or NGAL/uCr ratio did not correlate with reflux grade or laterality. Conclusion: The urinary level of NGAL and NGAL/uCr ratio might be a surrogate non invasive, reliable tool to distinguish hydronephrosis. http://rmm.mazums.ac.ir/article-1-50-en.pdf 2013-10-26 10 16 10.18869/acadpub.rmm.1.2.10 NGAL Antenatal hyronephrosis Vesicoureteral reflux Neonate Creatinine Urine Hamid MohammadJafari hamidmjaafari@yahoo.com 1 Department of Pediatrics, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. AUTHOR Alireza Rafiei Rafiei1710@gmail.com 2 Molecular and Cell Biology Research Center, Department of Immunology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran AUTHOR Mohammad Abedi 3 Medical student, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran AUTHOR Abdolrasul Aalaee 4 Department of Radiology and Nuclear Medicine, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran AUTHOR Araz Mohammad Mirabi 5 Molecular and Cell Biology Research Center, Department of Immunology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran AUTHOR Ehsan Abedi 6 Department of Radiology and Nuclear Medicine, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran AUTHOR
OTHERS_CITABLE Over-expression of CXCR4, a stemness enhancer, in human blastocysts by low level laser irradiation The key role of chemokine receptor CXCR4 in the maintenance of stemness property of stem cells has been shown recently. The low level laser irradiation (LLLI) is being used currently in a wide variety of clinical cases as a therapeutic tool for wound healing, relieving pain and destroying tumor cells. The aim of this study was to evaluate the effect of LLLI mimicking low level laser therapy (LLLT) on the expression level of CXCR4 gene a few hours after irradiation on human blastocysts. After the development of human embryos to the first grade blastocyst stage, they were irradiated with a low power Ga-Al-As laser at a continuous wavelength of 650 nm and a power output of 30 mW. The total RNA of the irradiated blastocysts and control groups were isolated in groups of 1x2 J/cm2, 2x2 J/cm2, 1x4 J/cm2 and 2x4 J/cm2 LLLI. Specific Real-Time PCR primers were designed to amplify all the two CXCR4 isoforms yet identified. RNA amplifications were done for all the groups. We showed for the first time that LLLI makes the human blastocysts to increase the expression level of CXCR4 a few hours after irradiation. Moreover, it was shown that two irradiation doses with one day interval can cause a significant increase in CXCR4 expression level in human blastocysts. This study revealed that LLLI could be a proliferation motivator for embryonic cell divisions through enhanced over-expression of CXCR4 level. http://rmm.mazums.ac.ir/article-1-39-en.pdf 2013-10-22 17 22 10.18869/acadpub.rmm.1.2.17 CXCR4 Human blastocysts Low-level laser irradiation (LLLI) Proliferation Stemness Mohammad Hossein Tahmasbi Soleim_m@modares.ac.ir 1 Cellular and Molecular Research Centre, Tehran University of Medical sciences (TUMS), Tehran, Iran AUTHOR Mohammad Shafiee 2 Cellular and Molecular Research Centre, Tehran University of Medical sciences (TUMS), Tehran, Iran AUTHOR Mohammad Taghi Joghataei 3 Omics Research Center, Golestan University of Medical Sciences, Gorgan, Iran AUTHOR Seyed Amir Yazdanparast 4 Cellular and Molecular Research Centre, Tehran University of Medical sciences (TUMS), Tehran, Iran AUTHOR Seyed Akbar Moosavi 5 Department of Allied Health, Tehran University of Medical Sciences (TUMS), Tehran, Iran AUTHOR Mina Jafarabadi 6 Reproductive Health Research Center, Tehran University of Medical Sciences (TUMS), Tehran, Iran AUTHOR Masoud Soleimani 7 Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran AUTHOR
OTHERS_CITABLE A simplified protocol for producing Taq DNA polymerase in biology laboratory Background: Taq DNA polymerase is a very important enzyme for molecular biological studies such as DNA amplification and DNA sequencing by the PCR. It is a standard enzyme that is used in 90% of molecular biology labs today. The aim of this study was to produce Taq DNA polymerase enzyme in E. coli by a reliable, practical, simple and low cost method. Materials and Methods: In this study, the Taq gene was amplified from the genomic DNA of Thermus aquaticus and cloned into pTrc99A vector. Recombinant plasmid is expressed in E. coli strain TOP10. Product protein is extracted and purified. Expression of gene was analyzed by SDS-PAGE and gene amplification. Results: SDS-PAGE showed an approximately 94 KDa protein. The density of protein bands in agarose gel electrophoresis indicated that the purified enzyme is more active than the non purified one. Conclusion: The protocols described in this paper lead to the production of pure and active enzyme that can be applied in both teaching and research laboratories. http://rmm.mazums.ac.ir/article-1-40-en.pdf 2013-10-23 23 26 10.18869/acadpub.rmm.1.2.23 Taq polymerase expression Purification Touraj Farazmandfar 1 Faculty of Advanced Medical Science Technologies, Golestan University of Medical Sciences, Gorgan, Iran AUTHOR Alireza Rafiei Rafiei1710@gmail.com 2 Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran AUTHOR Mohammad Bagher Hashemi-Sotehoh 3 Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran AUTHOR Reza Valadan 4 Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran AUTHOR Mohammad Alavi 5 Sari Agricultural sciences & Natural Resources University, Sari, Iran AUTHOR Fatemeh Moradian 6 Sari Agricultural sciences & Natural Resources University, Sari, Iran AUTHOR
OTHERS_CITABLE Effect of smoking on IFN-γ and IL-10 cytokines level in gingival crevicular fluid and clinical parameters of periodontium in Babol city Background: Periodontitis is an inflammatory disease of tooth-supporting tissues. The present study was conducted to evaluate Interleukin-10 (IL-10) and Interferon gamma (IFN-γ) levels in gingival crevicular fluid (GCF) of patients with chronic periodontitis. Materials and Methods: This case-control study was carried out on 60 men including 30 smokers and 30 age-matched non-smokers. The assessment of periodontal health was performed by using dental plaque, Barnett gingival bleeding and probing pocket depth (PPD) index. Cytokines level in the GCF evaluated by Enzyme linked Immunosorbent Assay (ELISA). Results: The mean value of dental plaque index showed no significant difference between the two groups (P=0.1). Although gingival bleeding index was higher in control compared to the case group, the difference was not significant (P=0.08). The mean probing pocket depth (PPD) was lower in the case that the control group (P=0.02). The mean of IL-10 and IFN-γ levels were 1.25 (pg/ml) and 0.82 (pg/ml) in smokers, and 1.22 (pg/ml) and 0.75 (pg/ml) in non-smokers (P>0.05), respectively. Conclusion: The findings showed that IL-10 and IFN-γ levels in the GCF of smokers were higher compared with non-smokers with chronic periodontitis, however, the difference was not significant. Further investigations including the evaluation of the other inflammatory mediators are required. http://rmm.mazums.ac.ir/article-1-41-en.pdf 2013-10-22 27 32 10.18869/acadpub.rmm.1.2.27 Chronic Periodontitis Smoking Interleukin-10 IFN-γ GCF. Ghorban Maliji azadmehr2010@gmail.com 1 Department of immunology and microbiology, Babol University of Medical Sciences, Babol, Iran AUTHOR Sina Jafari aazadmehr@qums.ac.ir 2 Department of immunology, Qazvin University of Medical Sciences, Qazvin, Iran AUTHOR Abbas Azadmehr azadmehr2012@gmail.com 3 Department of immunology and microbiology, Babol University of Medical Sciences, Babol, Iran AUTHOR Seyed Ehsan Moosavi ghmaliji@yahoo.com 4 Student research committee, dental school, University of Medical Sciences, Babol, Iran AUTHOR Ebad Taheri azadmehr2012@gmail.com 5 Dental School, University of Medical Sciences, Babol, Iran AUTHOR Ehsan Maliji ghmaliji@yahoo.com 6 Dental School, University of Medical Sciences, Babol, Iran AUTHOR
OTHERS_CITABLE 3D study of capillary network derived from human cord blood mesenchymal stem cells and differentiated into endothelial cell with VEGFR2 protein expression New blood forming vessels are produced by differentiation of mesodermal precursor cells to angioblasts that become endothelial cells (ECs) which in turn give rise to primitive capillary network. Human cord blood (HCB) contains large subsets of mononuclear cells (MNCs) that can be differentiated into endothelial-like cells in vitro. Human mononuclear progenitor cells were purified from fresh umbilical cord blood by the expression of CD34 and FLK-1 antigens expressed in both angioblasts and hematopoetic stem cells. The HCB derived mesenchymal stem cells (MSCs) can be differentiated into adipocyte, osteocyte, chondrocyte and ECs. In this study, the differentiation of human cord blood mesenchymal stem cells (hCBMSCs) into endothelial-like cells was induced in the presence of vascular endothelial growth factor (VEGF) and insulin-like growth factor (IGF-1). The differentiated ECs were then examined for their ability to express VEGF receptor-2 (VEGFR2) and von Willebrand factor (vWF). These cells were adopted to grow, proliferate and develop into a capillary network in a semisolid gel matrix in vitro. The capillary network formation in each well of 24-well plate was found to be 80% in presence of VEGF (40 ng/ml) and IGF-1 (20 ng/ml) of culture media, suggesting that the capillary network formation is associated with endothelial-like cells derived from hCBMSCs by expression of their markers. http://rmm.mazums.ac.ir/article-1-38-en.pdf 2013-10-22 33 38 10.18869/acadpub.rmm.1.2.33 Human umbilical cord blood (HUCB) Mesenchaymal stem cell (MSC) Angiogenesis Differentiation IGF-1 Mohammad Hossein Tahmasbi 1 1Cellular and Molecular Research Center, Tehran University of Medical Sciences (TUMS), Tehran, Iran AUTHOR Mohammad Taghi Joghataei Joghataei@Iums.ac.ir 2 1Cellular and Molecular Research Center, Tehran University of Medical Sciences (TUMS), Tehran, Iran AUTHOR Masoud Soleimani 3 Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran AUTHOR Seyed Akbar Moosavi 4 Department of Allied Health, Tehran University of Medical Sciences (TUMS), Tehran, Iran AUTHOR Seyed Amir Yazdanparast 5 1Cellular and Molecular Research Center, Tehran University of Medical Sciences (TUMS), Tehran, Iran AUTHOR Farah Zaker 6 1Cellular and Molecular Research Center, Tehran University of Medical Sciences (TUMS), Tehran, Iran AUTHOR
OTHERS_CITABLE Comparison of TNF-α and TGF-β1 level in radicular cyst and odontogenic keratocyst fluid and its association with histopathological findings Background: TNF-α is a multifunctional proinflammatory cytokine and TGF-β1 is a secretory protein controlling epithelial proliferation and differentiation. Keratocyst presents an aggressive behavior and a growth mechanism different from that of radicular cyst. Aim: In this line, the present study aimed at evaluating TNF-α and TGF-β1 level and its association with histopathological findings in the two odontogenic lesions of different origins. Materials and Methods: In this case-control study, aspirated fluid of 15 cases of radicular cyst and 15 cases of keratocyst were investigated using ELISA method. The grade of inflammation and the mean number of blood vessels in three microscopic fields were provided with a magnification of 40 times on microscope slides. T-test, x2, Mann Whitney, and Pearson correlation tests were used for the comparison of TNF-α and TGF-β1 levels in the mentioned lesions and the association between cytokine levels and grade of inflammation and angiogenesis. Results: TNF-α and TGF-β1 were observed in aspirated fluid of all radicular cysts and keratocysts. Levels of TNF-α and TGF-β1 were found to be 6.72 ± 2.985 and 5.882 ± 2.985 respectively in radicular cyst fluid and 24.759 ± 94.849 and 63.38 ± 30.069 in keratocyst fluid however, no statistically significant difference was observed in terms of TNF-α (P=0.450) increasing trend in TNF-α level in radicular cyst and keratocyst was accompanied by increased inflammation and angiogenesis (P<0.001 and P=0.001). Conclusion: TNF-α and TGF-β1 are involved in the pathogenesis of radicular cyst and keratocyst. TGF-β1 level was higher in radicular cyst when compared with keratocyst however, TNF-α level was similar in the two lesions. A positive correlation was found between TNF-α level and grade of inflammation and angiogenesis. http://rmm.mazums.ac.ir/article-1-46-en.pdf 2013-10-22 39 43 10.18869/acadpub.rmm.1.2.39 Radicular cysts Odontogenic keratocyst TNF-α TGF-β1 ELISA Safoura Seifi sf_seify@yahoo.com 1 Department of Oral and Maxillofacial Pathology, School of Dentistry, Babol University of Medical Sciences. Babol, Iran AUTHOR Mohammad Mehdizadeh DR.rmohammad@yahoo.com 2 Department of Oral and Maxillofacial Surgery, School of Dentistry, Babol University of Medical Sciences. Babol, Iran AUTHOR Ghorban Maliji ghmaliji@yahoo.com 3 Department of Immunology, Faculty of Medicine, Babol University of Medical Sciences. Babol, Iran AUTHOR Zahra Sadat Korsavi zahra.korsavi@gmail.com 4 Dentist, School of Dentistry, Babol University of Medical Sciences Iran. Babol, Iran AUTHOR Kamran Nosrati nnosrati@yahoo.com 5 Department of Oral and Maxillofacial Surgery, School of Dentistry, Babol University of Medical Sciences.Babol, Iran AUTHOR
OTHERS_CITABLE The hormonal milieu in primary breast cancer: A correlation between steroid receptors and serum estradiol, progesterone and prolactin Background: The female breast is subjected to a lifetime of hormonal controls, whose effect is evident at the time of menarche and during the menstrual cycle, pregnancy and lactation. Studies have reported multiple risk factors for breast cancer, some of which are a reflection of hormonally mediated events. The steroid receptors are also served as prognostic factors for evaluating status of malignant tumor of the breast. Estrogen receptor (ER) and progesterone receptor (PgR) formation are influenced by Estrogen and progesterone concentration. The aim of this study was to clarify the hormonal milieu of the breast cancer such as Estradiol (E2), Progesterone (Pg), Prolactin (PRL) and their correlation with prognostic factors like ER, PgR. Materials and Methods: In this study we examined fourthy four samples removed from patients with primary breast cancer by radical mastectomy. The specimens include thirty six malignant and eight benign breast tissues. Blood samples were also obtained before surgery. steroid receptors was assayed by the method of single-point dextrane-coated charcoal (DCC), hormones by radioimmunoassay. Results: The results demonstrated that serum prolactin was higher in patients with benign and malignant tumors than that of normal (progesterone<0.05). It has been observed that the frequency of PgR+ tumors was higher in patients with serum E2 more than 10 pg/ml than those of less than 10 pg/ml where as, when comparison was made between patients with serum Pg>1.5 ng/ml and those of <1.5 ng/ml, it was found that frequency of PgR+ tumors was lower in the formers than the latters. Conclusion: It can be concluded that estrogen, up-regulated and progesterone down-requlate the induction of PgR and the breast tumors probably produce high levels of prolactin. http://rmm.mazums.ac.ir/article-1-42-en.pdf 2013-10-22 44 47 10.18869/acadpub.rmm.1.2.44 Breast cancer ER Estrogen PgR Progesterone Prolactin Mehryar Zargari zargari.mehryar@gmail.com 1 Department of Biochemistry, Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. AUTHOR