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Abstract Background and objectives: Breast cancer is the most prevalent one in women. Some of the common causative factors are genetic background, nutritional and environmental factors. Viruses are believed as a risk factor in this cancer, too. Recent studies reported that Human Papillomaviruses can be one of the possible risk factors of breast cancer. This study focused on investigation of the papillomavirus genome in tissues of breast cancer in Golestan province, Iran. Material and Methods: This descriptive analytical study was done from 2005 until 2008. The Samples were obtained from women admitted to the hospitals in Gorgan and Gonbad cities. All breast biopsy or mastectomy tissues were confirmed by the pathologists for breast cancer. DNA was extracted and PCR done by using general primers (GP5 + / GP6 + and MY09/MY11) for detection of papillomavirus genomes. Results: The Subjects are 231 patients aged 47± 12/72, the youngest 20 and the oldest 84. They are from Gorgan (N=122)and Gonbad (N=109) The result Shows That The Subjects Suffer from infiltrating ductular Carcinoma(31.4%), infiltrating duct Carcinoma (60.1%)and intraductal Lobular Carcinoma (4.3%) and The rest from other kinds of Cancer. Papilloma Virus genome is not found in These Samples. Conclusion: Based on paradoxical results from different parts of the world, upon the presence or absence of papillomavirus genome in breast cancer samples, to show the role of this virus in the development of breast cancer more studies are needed. Key words: Breast Cancer, Papillomaviruses, Golestan Province

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Abstract Background and objectives: Beta-lactamase enzymes are the most causes of resistance to antibiotics among gram-negative bacteria. Nowadays, Infections due to ESBLs are being increased throughout the world and is considered as a new burden to the health systems. This study aimed at determining the sensitivity pattern of E.coli isolates to beta-lactam antibiotics, and investigating the presence of blaCTX-M, blaTEM, and blaSHV genes in the urine samples. . Material and Methods: In this study, 244 E.coli isolates were screened in 2009-2010. The antibiotic susceptibility of E. coli isolates were determined by disc-diffusion method. Antimicrobial agents tested were cefoxatime, ceftazidime, imipenem, nalidixic acid, and ciprofloxacin. The combined disc test was used to confirm the results. The results were compared to the Clinical and Laboratory Standards Institute (CLSI) and ESBL positive isolates were further investigated for the presence of blaCTX-M, blaTEM, and blaSHV genes by PCR. Results: Of 244 E. coli isolates, 116 (47.1%) are resistant to Ceftazidime, and 96 (39.2%) to cefoxatime. Also, 109 (44.3%) isolates are ESBL positive. blaCTX-M, blaTEM, and blaSHV genes are found among 95 (87.1%), 75 (68.8%), and 77 (70.6%) ESBL positive isolates, respectively. Forty (36.6%) isolates have all three genes, while 68 (62.3%) include blaTEM and blaSHV genes. Moreover, 61 (55.9%) isolates carry blaCTX-M and blaTEM genes, and 54(49.5%) have blactx-M and blashv. Conclusion: Regarding the high frequency of resistance to the third generation cephalosporin antibiotics, precise antibiogram testing is highly recommended before any antibiotic prescription in cases of infections with ESBL producing microorganisms. Key words: ESBL Escherichia coli blaCTX-M blaTEM blaSHV.

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Background and Objectives: rapid, accurate and cost effective diagnosis of infectious and non infectious diseases is an essential step for treatment process. Nowadays, in Line with scientific progression in molecular biology, genetics and biochemistry which are based on biotechnology and genetic engineering aspects, new branch of medicine entitled molecular medicine is being derived. It can be helpful in three areas of diagnosis, prophylaxis and treatment. This new branch is going to identify further complexity of diseases and to present efficient solutions for growing health criteria. Therefore, updating and being familiar with the new procedures related to diagnosis, prophylaxis and therapy are necessary for our society. In this paper, we are trying to introduce NASBA technology which has a high potential, at genome level, in recognizing specific characteristic and unique genetic markers of microorganisms. This technology has numerous benefits for easy detection of infectious diseases such as tuberculosis. Furthermore, we review the methods of tuberculosis detection.

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Abstract Background and objectives: Mumps virus is one of the first known causative agents of meningitis in children. On-time diagnosis is the first step in treating meningitis. We aimed to evaluate Mumps virus meningitis in children in Gorgan, Iran Material and Methods: CSF and blood samples were taken from children with meningitis, Jun 2008 till Sep 2010. For 40 samples with negative bacterial culture, Extraction of viral RNA was carried out and Real-time PCR was performed for detection of Mumps virus. Demographic, clinical, biochemical and cytological data were collected. We run SPSS version 18 to analyze the data, using Chi Square (p<0.05). Results: three (7.5 %) samples have Mumps virus, two boys and one girl. All three positive cases have 0.5-1 degrees Celsius fever and vomiting but no bulging fontanel. They have not Kernig, Rodor, Brudzinski’s sign, hepatosplenomegaly, lymphadenopathy, pharyngitis and rash. ESR is higher than normal in all positive cases and CRP is positive in two cases. Protein of CSF in one case is higher than normal range. Conclusion: meningitis is an emergency condition therefore, molecular diagnostic techniques are recommended for early diagnosis and intervention. Key words: meningitis, mumps virus, cerebrospinal fluid, Real-Time PCR

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Abstract Background and objectives: Identification and monitoring of multidrug-resistant Mycobacterium tuberculosis strains (MDR) is highlighted by the high risk of their spreading in different areas. Prevalence of these strains was evaluated in Golestan province in northeast of Iran. Material and Methods: Drug susceptibility testing to Isoniazid and rifampin was carried out for 148 clinical samples that had grown in Mycobacteria growth indicator tube (MGIT) system, according to the manufacturer's instructions (Becton-Dickinson, USA). The association of drug resistance frequency with demographic characteristics and growth time were investigated. The appropriate statistical tests, X2 and student T- test were performed for comparison of these variants. A p value>0.05 was considered significant in all cases. Results: The turnaround time required for growth of Mycobacterium tuberculosis in MGIT system was between 2 to 55 days (mean 16.3±10.4 days). Of all samples studied, 17.6% and 3.4% were resistant to Isoniazid and rifampin, respectively, and 3.4% (5 samples) were MDR (CI 95% 1-6%). The turnaround time required for determining MDR cases was 9.6 days. No statistically significant association was found between the resistance to the drugs and none of the factors including sex, age, type of clinical sample, and positivity of the smear. Conclusion: The prevalence of MDR in the studied region was determined to be 3.4% which is similar to the country-wide evaluations. The turnaround time for Mycobacterium growth and anti drug susceptibility result can be shortened by MGIT method. Key words: Mycobacterium tuberculosis, Mycobacterium Growth Indicator Tube, Multidrug Resistant

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Abstract Background and objectives: Hepatitis B virus infection is a major health problem in worldwide. The prevalence of Occult and chronic HBV in hemodialysis patients is higher than standard in developing countries. People with occult HBV are negative for HBV surface antigen (HBsAg) but positive for HBV-DNA. We aimed to evaluate occult hepatitis B infection in patients under hemodialysis in Panje-Azar hospital in Gorgan. Material and Methods: In this study, taken place from 2009 to 2010, the participants were 100 hemodialysis patients with administration of complete HBV vaccination with negative test for HBsAg. After preparing 10 milliliter blood sample, HBV DNA testing was performed by polymerase chain reaction (PCR). Result: The mean age of the patients is 54.60 years. They are male (48%) and female (52%). They have been under hemodialysis for 48 months, averagely. There has not been any HBV-DNA in HBsAg negative patients under hemodialysis. The rate of occult hepatitis B infection in these end stage renal disease (ESRD) patients was zero. Conclusion: Results indicate that there is no any occult HBV infection in ESRD patients under hemodialysis in Gorgan, which is similar to some studies. The results could be justified by complete vaccination of the patients. Key words: Occult Hepatitis B, Hemodialysis, HBsAg, Gorgan

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Abstract Background and objectives: The presence of pathogenic bacteria and the factors causing food spoilage are the great challenge for public health. Attention to natural additives instead of chemical preservatives resulted in conducting several studies on plant essential oil and extracts. We aimed at evaluating the antibacterial effect of carboxymethyl cellulose coating enriched by Zataria multiflora essential oil and grape seed extract on rainbow trout meat. Material and methods: In this study, two concentrations of Zataria multiflora essential oil (1% and 2%) and two concentrations of grape seed extract (0.5% and 1%) were used both alone and in combination with Carboxymethyl cellulose coating. Antibacterial effect of these treatments was evaluated by enumeration of bacteria in special culture media. Results: The results obtained in this study demonstrate that Zataria multiflora essential oil in combination with grape seed extract significantly can decrease the number of bacteria and delay the spoilage of the samples (p<0.05). Conclusion: Coating enriched by Zataria multiflora and grape seed extract can properly delay the growth of spoilage microorganisms and prolong the shelf life of meat products. Key words: Carboxymethyl cellulose coating, Zataria multiflora essential oil, Grape seed extract, Microbial flora

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Abstract Background and objective: Hepcidin is a cystein-rich antimicrobial peptide, which is secreted by the liver. It fights against wide spectrum of bacteria, viruses and fungi and it is a major regulator of iron homeostasis. Today, scientists have made many efforts on the production of hepcidin. Baculovirus expression system is one of the best eukaryotic expression systems for production of recombinant hepcidin and production of the recombinant vector is one of the most important steps in this expression system. Material & Methods: First, the total RNA was separated from HepG2 cell line as a source of hepcidin expression. Then, after synthesis of total cDNA, human hepcidin sequence was amplified, using specific primers by PCR method. Next, hepcidin sequence was cloned into pTZ57R/T vector. After digestion of recombinant vector using ECoRI and BamHI restriction enzymes, recombinant pFastBac HT B vector containing human hepcidin cDNA was produced. Results: Coding sequence of human hepcidin is correctly cloned into pTZ57R/T vector and sub cloning into pFastBac HT B vector is performed successfully. The presence of a clear band near 274 bp resulted from PCR amplification and restriction enzyme are the confirmation of the cloning of human hepcidin. Conclusion: According to our knowledge, the present study is the first work that focuses on recombinant vector containing coding sequence of human prohepcidin. This recombinant vector can be used for human hepcidin production. Key words: Vector, Hepcidin, Iron

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Abstract Background & Objective: Hepatitis E virus is one of the most common causes of acute infection in adults. Pregnant and transplant patients are more in risk of HEV infection. Fecal-oral is the main route of HEV transmission but recently transmission by blood transfusion has been observed. This study was aimed to determine the prevalence of HEV-Ab in hemodialysis patients in Gorgan, Iran. Material and Methods: In this cross-sectional descriptive study, we investigated 150 hemodialysis patients of Panje Azar hospital in Gorgan. These patients were evaluated for the presence of HEV total Ab by ELISA method. Results: of 150, 6 patients (4%) are positive for HEV-Ab. There has been no significant relation between anti HEV Ab and variables such as age, gender, ethnicity, duration and number of hemodialysis in a week and (P>0.05). Conclusion: This study, which is the first report from this area, show that the lower prevalence of anti HEV Ab in hemodialysis patients in comparison with pregnant and childbearing age women. Keywords: Hepatitis E Hemodialysis Elisa Gorgan

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Abstract Background and Objective: Resistance to antiretroviral agents is a significant concern in clinical management of HIV-infected individuals. Resistance is the result of mutations that develops in the viral protein targeted by antiretroviral agents. Material and Methods: In this cross-sectional study, the blood samples of 40 HIV-positive patients were collected. Twenty of them were drug-naïve and the rest were under treatment for at least one year by antiretroviral agents. Virus genome was extracted from patient's plasma with high-pure-viral-nucleic-acid kit. Then, by means of reverse-transcriptase and specific primers of protease genes were amplified and sequenced. Sequences of genes, drug- antiretroviral- resistant mutations and subtypes were determined using Stanford University’s HIV-drug-resistance databases. Results: Drug-naive patients show 15% resistance to nucleoside-reverse-transcriptase inhibitor (NRTI) and 20% resistance to non-nucleoside-reverse-transcriptase inhibitor (NNRTI). Anti-protease resistance is not observed in any patients. In under treatment patients, drug resistance to NNRTI (25%) is more than drug resistance to NRTI (20%) and the rate of drug resistance to protease inhibitor is 5%. Conclusion: Our findings show a high prevalence of drug-resistant mutations in Iranian-drug-naïve-HIV-infected patients. But in under treatment individuals, the rate of drug resistance is less than previous studies. Keywords: HIV Nucleoside Inhibitor Non-Nucleoside Inhibitor Protease Inhibitor

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Abstract Background and Objective: Parasitic infection is one of the major health problems in the world. This study aimed at comparing the accuracy of two methods of direct examination and Formalin-Ether to detect the presence of parasitic infection among health-card applicants in Shahroud city, 2011. Material and Methods: This cross-sectional study was conducted on 801 patients seeking health-card. From each patient, three consecutive stool samples were taken and investigated, using direct examination and formalin-ether method. Results: The use of formalin-ether method in recognizing the parasitic infection specially giardia lamblia and entamobea coli is more than the direct method. Conclusion: The formalin-ether method is a more sensitive method than the direct method. But in circumstances that is urgency to respond or aims to see the shape of trophozoite, the use of direct method is recommended. Keywords: Parasitic Infections Health Card Direct Method Formalin-Ether

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Abstract Background and Objective: Respiratory tract infections (RTI) are the most common infectious disorders, worldwide. About 80%-90% of RTI are caused by four viruses such as Adenoviruses, 51 serotypes have been introduced so far. The aim of this survey was to evaluate the frequency of Adenovirus in respiratory infected patients by PCR method in Golestan province, Iran. Material and Methods: This descriptive cross-sectional study was conducted on 400 patients with clinical diagnosis of flu-like respiratory infection, 2010-2012. In addition to collecting demographic and clinical data, nasopharyngeal swabs were taken and transferred to the virology laboratory in viral transport medium (VTM), and evaluated by PCR method for Adenovirus after genomic extraction. Using SPSS v.11 software, we analyzed the data. Results: Thirty-seven (9.2 %) were positive for Adenovirus. No significant correlation was found between being positive for Adenovirus and the variables such as age, gender and season. Clinical signs were coughing (27 73%), body pain (25 67.6%), and fever (24 64.9%). Thirty-five of the patients (94.5%) had at least one symptom. Conclusion: Our findings are consistent with other research conducted in Iran and other countries. There is a significant correlation between Adenovirus infection and clinical symptoms. Keywords: Respiratory Infection, Adenovirus, PCR, Golestan, Iran

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Abstract Background and Objective: The Plasmodium falciparum EBA-175, via Sialic acid dependent glycophorin A, binds to red blood cells and thus plays a critical role in cell invasion. Some part of second allele in its gene encoding in FCR-3 (Section F) and CAMP (Section C) can be found. This study aimed to determine the prevalence of Plasmodium falciparum EBA-175KD alleles in southeastern Iran. Material and Methods: In this cross-sectional study, using polymerase chain reaction Nest (Nested-PCR) with specific primers was used for the two parts of the EBA-175 gene to be proliferated. Ninety–four microscopic positive blood samples from individuals infected by Plasmodium falciparum were obtained from four different locations in southeastern Iran. Results: Of 94 positive samples, 88 were antigen EBA-175KD. Genotype CAMP (714 bp) and FCR-3 (to 795 bp), respectively, in 31 (32.97 %) and 49 (52.12 %) were found. Eight samples have both FCR-3 and CAMP. Conclusion: Both of EBA-175KD dimorphic genes were found. The frequency of FCR-3 allele was higher in the South East of Iran. Thus, this pattern can be considered in making Plasmodium falciparum vaccines for this area. Key words: Plasmodium Falciparum Erythrocyte Binding Antigen-175 South-East of Iran

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Abstract Background and Objective: The emergence of a novel H1N1influenza A virus of animal origin with transmissibility from human to human poses pandemic concern. Current subtypes of Seasonal influenza A viruses spread in human are influenza A H1N1 influenza A H3N2 and influenza type B viruses. The aim of this study was to determine current strains of the H3N2 and new H1N1 subtypes of influenza A virus from patients suspected influenza infection in 2009 flu pandemic in Golestan province, Iran. Material and Methods: In this descriptive study, respiratory samples (n = 153) from patients with acute respiratory symptoms were collected in 2009 flu pandemic applied during 2009 pandemic influenza in Golestan province. After reverse transcription of extracted viral RNA, PCR was developed for both H1N1and H3N2subtypes using CDC specific primers. Results: The mean age of patients was 16.59. Of them 45.1% were male. Thirteen (8.49%) were infected with seasonal influenza H1N1 and 25(16.33%) with seasonal H3N2influenza. Conclusion: The rate of infection with seasonal H1N1and H3N2is similar to other studies reported from Iran, but lower than the rate reported from other parts of the world. Key Words: Influenza A Virus, H1N1, H3N2, RT-PCR, Iran

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Background and Objective: Cytomegalovirus (CMV), one of the most common opportunistic pathogens in patients infected with human immunodeficiency virus (HIV), can cause the diseases such as encephalitis, pneumonia, and chorioretinitis. This study aimed at molecular studying of CMV infection in individuals infected with the human immunodeficiency virus. Material and Methods: In this study, 50 venous blood samples from HIV-infected individuals were taken. Patients were divided into two categories: patients under treatment with and without antiretroviral drugs. Plasma were separated from blood samples and examined for the presence of cytomegalovirus genome by PCR. Material and Methods: this study was conducted on 50 blood samples from HIV-infected individuals, and plasma was separated and examined for the presence of cytomegalovirus genome by PCR. Patients were divided into two group of under treatment with and without antiretroviral drugs. Results: Of 50, 28 (% 56) were men and 22 (% 44) were women. CMV genome was identified in 8 samples (16%), and the molecular prevalence of CMV infection was 21.4% (n= 6) in males and 9.1% (n = 2) in females. Conclusion: Given the frequency of Cytomegalovirus Active Infection in HIV-infected individuals under antiretroviral therapy, we should be careful about the treatment of Cytomegalovirus Active Infection. Keywords: Active Infection, Cytomegalovirus, Human Immunodeficiency Virus, Shiraz, PCR

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Abstract Background and Objective: Lamivudine is the first orally available drug approved for treatment of chronic hepatitis B. Mutations at the YMDD and FLLAQ motifs in the domains of HBV polymerase gene contribute resistance to lamivudine. This study was aimed to determine the rate of YMDD and FLLAQ mutants in hepatitis B patients in Golestan Province, Iran. Material and methods: In this cross sectional study, 120 patients with chronic HBV infection were recruited. Of them, 55 were treated and 65 untreated with Lamivudine. HBV DNA extractions from plasma and polymerase chain reaction (PCR) were performed. For detection of Lamivudine mutants direct sequencing and alignment of products were applied using reference sequence from Gene Bank database. Results: the average age of patients was 36.31±10.07, which 35% of them were female and 65% were male. Mutations at the YMDD and FLLAQ motifs in the domains of HBV polymerase gene were detected in 12 of 55 patients (21.81%) treated with Lamivudine while no mutation was observed in in untreated patients. The YMDD and FLLAQ mutants were detected in 9.16% (11/120) and 0.83% (1/120) of chronic HBV patients, respectively. Conclusion: Usual HBV mutations, which play an important role in lamivudine resistance, detected in this study are similar to other studies. Key words: Hepatitis B Viruse, YMDD Mutation, Lamivudine, Iran.

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Abstract Background and Objective: Human Cytomegalovirus (CMV) is an important cause of congenital viral infection that can lead to serious diseases and complications in infants. Application of rapid, sensitive, and specific HCMV detection methods is necessary for congenital infection detection. We aimed to optimize the use of PCR and ELISA for detection of HCMV in infants. Material and Methods: PCR–ELISA was performed by using specific primers and probe for detection of the HCMV glycoprotein B gene. First, the extracted DNA from urine samples and controls were labeled by digoxigenin during DIG-labeling PCR. After that, Biotin-labeled probe captured the DIG-labeled PCR products. The probe-PCR product hybrid is immobilized on a streptavidin-coated Microtiter plate, and detection was confirmed by proxidase-conjugated anti-digoxigenin antibody, and calorimetric substrate. Results: The clinical Human CMV strains isolated from16 patients were detected by this method. The optimized PCR-ELISA method was able to detect less than100 copies of HCMV genome. There was no non-specific reaction. Conclusion: PCR-ELISA can be applied as a sensitive, specific and reliable method for Semi-quantitative CMV detection in clinical samples. Keywords: Cytomegalovirus, Glycoprotein B, PCR-ELISA, Semi-Quantitative

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Abstract Background and Objective: The bacteria living in the specific ecological conditions are among the most promising antimicrobial producers. This study aimed at isolating antimicrobial producing bacteria from soils contaminated with crude oil. Material and Methods: the samples were obtained from crude oil contaminated soils around Dezful located in Khuzestan province, Iran, and antimicrobial producing bacteria were isolated using disc diffusion and cross streak culture. Then, the best bacterium was selected and its antimicrobial potency was studied against indicator microorganisms. The isolate was also characterized based on biochemical properties and phylogenetic analysis. Results: based on the results, the highest antimicrobial activity of isolated bacterium was related to Candida albicans, Aspergillus niger, Bacillus subtilis, E. coli and Klebsiella pneumonia. An intermediate effect was determined against Serratia marcesens and Staphylococcus aureus, whereas no effect was observed against three strains of Enterococcus. Using biochemical characteristics and phenotypic traits, the isolate was identified as Alcaligenes faecalis. Conclusion: given that the isolate has broad spectrum activity against a various range of microorganisms and in comparison with some antimicrobial compounds produced by other Alcaligenes species, it seems the novelty of this antimicrobial compound. Keywords: Antimicrobial Compound, Oil Contaminated Soil, Alcaligenes faecalis

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Abstract Background and Objective: Highly Active Antiretroviral Therapy (HAART) can effectively prevent the progression of HIV-1 replication and increase life expectancy. There are numerous causes of treatment failure and the leading one is drug resistance. Thus, we aimed to determine the HIV RT gene drug resistance mutations in patients treated with antiretroviral medications. Material and Methods: In this cross - sectional study, venous blood was taken from 130 HIV-positive patients treated with antiretroviral medications. In order to determine drug resistance mutations, RT-PCR and PCR steps were performed using RT gene specific primers. Subtypes and mutations in the virus genome were determined using the Stanford HIV drug resistance sequence database. Results: In 122 treating patients, most of the major mutations were associated with nucleoside and non-nucleoside drugs. subtype A in 66.4%, subtype D in 26.2% and subtype B in 7.4% of the participants were reported. They were resistant to Nucleoside RT Inhibitor drugs (23.7%) and Non-Nucleoside RT Inhibitor drugs(30.3%). The highest were related to Nevirapine (21.3%) and Efavirenz (19.7%) and the lowest to both Tenofovir and Zidovudine (91.5%). Conclusion: The use of two nucleoside RT inhibitor drugs combined with one protease inhibitor drug could be effective in the treatment of HAART. Key words: HIV, Nucleoside RT Inhibitor, Non- Nucleoside RT Inhibitor

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Abstract Background and Objective: The M2 gene expressing the conserved protein in influenza virus can be used to make a single-dose vaccine with permanent immunity. Material and Methods: The mice were allocated to one case group immunized with pcDNA3-M2 and two control groups with pcDNA and PBS, in three dozes with interval of two weeks. Two weeks after the last injection, Cellular immunity was analyzed by MTT lymphocyte proliferation, interferon gamma (IFN-gamma) and interleukin 4 (IL-4) ratio assays. The remaining animals were challenged with PR8 virus. Results: The production rate of IFN8 and IL4 in pcDNA - M2 group was higher than that of control groups (P >0.0001). Given the results of lymphocyte proliferation, Stimulation index (SI) in vaccinated mice was significantly higher than that of control groups (P<0.05). In comparison with mortality rate of 100% in control groups , the animals Challenged with PR8 vaccine had a 50% fatal rate implying a high protection level for this vaccine. Conclusion: The pcDNA3-M2 Vaccine can be considered as a promising vaccine against influenza infections. Keywords: Influenza Virus, Gene Vaccine, M2 Protein