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Background and Objective: Hemophilia A is the most common type of this disease. For the treatment of hemophiliacs, the injectable concentrate of human factor VIII, derived from plasma and recombinant, is available in the market. Due to the risk of contracting infectious diseases due to blood transfusions or donated blood products, including plasma-driven factor VIII, recombinant factor VIII can be used, which is safe and has a higher purity than the blood-derived sample, while showing a longer half-life in the blood of consuming patients. The aim of this study is to evaluate the expression and synthesis of recombinant human factor VIII using a cell-free eukaryotic expression system as a new strategy for the synthesis of recombinant drugs. Methods: In this experimental study, pcDNA3 vector containing the gene fragment without B-domain of blood coagulation factor VIII was transformed into susceptible DH5a bacteria. After the multiplication of the bacteria and the increase of the target vector, the amplified vector was extracted at this stage and its quality and concentration were determined. Then CHO-K1 eukaryotic cells were cultured and cell extract was prepared from the density of more than ten million of their cells. The extracted vector was combined with cell extract and expression solution, and the expression process was carried out at 30°C and 37°C. The level of expression was evaluated by ELISA and dot blot techniques. Findings: Based on the ELISA technique adopted in this study, the expression level of recombinant human blood coagulation factor VIII in the base sample at a temperature of 30°C, sample 2 at a temperature of 30°C, base sample at a temperature of 37°C, and sample 4 of negative control at a temperature of 30°C were respectively reported as 275, 276, 273 and 0 ng/ml in a period of 20 hours. In the dot blot technique, the mentioned quantitative results were qualitatively confirmed by the color intensity of the spots. Conclusion: The results showed that the cell-free expression method can be used for the synthesis of recombinant factor VIII. Under the conditions used in this research for cell-free expression of recombinant factor VIII, volume ratio of 50/50 of CHO-K1 cell extract was suitable, and the basic expression solution at 30°C showed the highest concentration of the product. Due to the use of CHOK1 eukaryotic cell extract, the synthesized recombinant human factor VIII has post-translational modifications, which are very important for the functionality of this protein in the human body. [ABSTRACT FROM AUTHOR]

Background: Klebsiella pneumoniae is a clinically relevant pathogen that has raised considerable public health concerns. This study aims to determine the presence of beta-lactamase genes and perform molecular genotyping of multidrug-resistant (MDR) K. pneumoniae clinical isolates., Methods: Clinical isolates of MDR K. pneumoniae were collected from educational hospitals affiliated with Babol University of Medical Sciences. The isolates of K. pneumoniae were identified through standard microbial and biochemical tests. Antibiotic resistance was assessed using disk diffusion, modified Hodge test (MHT), combined disk, and polymerase chain reaction (PCR) methods. Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR was performed for molecular typing., Results: A total of 42 MDR K. pneumoniae isolates were obtained from various clinical specimens. The highest antibiotic resistance was observed for ampicillin (100%), while the lowest resistance was noted for amikacin (19.04%). The MHT indicated that 38.09% of K. pneumoniae isolates produced carbapenemase enzymes. Metallo-beta-lactamase (MBL) production was found in 54.76% of isolates. Molecular detection of beta-lactamase genes revealed the presence of bla NDM (21.42%), bla KPC (42.85%), bla TEM (76.19%), bla SHV (47.16%), and bla CTX-M (80.95%) genes. ERIC-PCR molecular typing identified seven distinct genetic patterns among the isolates., Conclusions: This investigation demonstrates the high resistance levels of K. pneumoniae strains. The beta-lactamase genes with the highest and lowest frequencies correspond to bla CTX-M and bla NDM genes, respectively. ERIC-PCR dendrograms suggest a common origin for K. pneumoniae clinical isolates and the propagation of similar clones within hospital wards. These findings indicate that K. pneumoniae isolates are highly virulent, necessitating the development of more effective resistance-fighting techniques and gene transfer research., Clinical Trial Number: Not applicable., Competing Interests: Declarations. Ethics approval and consent to participate: This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of Islamic Azad University - Ayatollah Amoli Branch; Amol, Iran (2022-09-11/ IR.IAU.AMOL.REC.1401.087). Informed consent was obtained from all individual participants included in the study. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Antibiotic resistance is a significant global health concern, leading to increased morbidity, mortality, and health care costs. Integrons are genetic elements that could acquire and express gene cassettes, including those that confer antibiotic resistance. This comprehensive study focused on the distribution of integrons and their gene cassettes in clinical isolates. This study explored the structure and classification of integrons with particular emphasis on Class I, II, III, and IV integrons. It also discussed the role of integrons in antibiotic resistance. The findings of this study contribute to a better understanding of the mechanisms underlying antibiotic resistance and provide valuable insights for developing strategies to combat this public health crisis.

Gingival-derived mesenchymal stem cells (GMSCs) stand for a unique source of mesenchymal stem cells (MSCs) isolated from a neural crest origin with potential application in regenerative medicine. However, there are some limitations to the usage of these cells in clinical cell therapy such as reduced cell number and undesirable differentiation of the cell throughout frequent passages. Nowadays, studies have applied manipulation strategies to improve MSCs' effectiveness in clinical therapy. Among all of the materials used for this purpose, there is a growing trend for the use of biomaterials such as probiotic extracts or their conditioned media due to their lower toxicity. In the present study, we utilized extracts from Lactobacillus reuteri and Lactobacillus rhamnosus to assess their potential to enhance the function of GMSCs. We compared the effectiveness of these bacterial extracts to determine their relative efficacy. Bacterial extracts of two lactic acid bacteria were prepared using an ultrasonic homogenizing device. The impact of these bacterial extracts on GMSCs was evaluated through Alizarin Red and Oil Red O staining, cell counting by Trypan Blue staining, and real-time polymerase chain reaction. The findings of our study indicate that the administration of 50 μg/mL L. rhamnosus extract resulted in a greater enhancement of stemness marker expression, osteogenic differentiation, and proliferation of GMSCs compared with an equivalent concentration of L. reuteri extract. Neither of these bacterial extracts revealed any effect on the differentiation of the GMSCs into the adipogenic lineage. These findings suggest that L. rhamnosus extract could be more effective at promoting GMSCs' efficacy in tissue engineering and regenerative medicine.

The vagina hosts a community of microorganisms known as the vaginal microbiota. This community is relatively stable and straightforward, with Lactobacillus species being the most dominant members. The vaginal microbiota has various functions that are essential for maintaining human health and balance. For example, it can metabolise dietary nutrients, produce growth factors, communicate with other bacteria, modulate the immune system, and prevent the invasion of harmful pathogens. When the vaginal microbiota is disrupted, it can lead to diseases and infections. The observed disturbance is distinguished by a reduction in the prevalence of Lactobacillus and a concurrent rise in the number of other bacterial species that exhibit a higher tolerance to low oxygen levels. Gynecologic cancers are a group of cancers that affect the female reproductive organs and tissues, such as the ovaries, uterus, cervix, vagina, vulva, and endometrium. These cancers are a major global health problem for women. Understanding the complex interactions between the host and the vaginal microorganisms may provide new insights into the prevention and treatment of gynecologic cancers. This could improve the quality of life and health outcomes for women., (© 2024. The Author(s).)

Background: Coagulation factor VIII (FVIII) is applied for spontaneous hemorrhaging inhibition and excessive bleeding after trauma in patients with hemophilia A. High-quality human recombinant factor VIII (rFVIII) has been produced relatively in large quantities in cultured mammalian cells. NS0 is one of the most common mammalian cell lines for therapeutic protein production. Production of rFVIII has increased due to low FVIII expression levels and rising demand for hemophilia A prophylactic treatment. Several methods have been developed to prevent cell cycle progression in mammalian cells for increased recombinant protein yields., Objective: The aim of the study was to investigate the level of recombinant BDD-FVIII expression in NS0 mouse myeloma cells. Additionally, the study aimed to determine the effects of chemical drugs, Mitomycin C, Lovastatin, and Metformin on the secretion of FVIII through cell cycle arrest., Materials and Methods: We cultured NS0 cells and transfected them with the 2 μg pcDNA3-hBDDFVIII plasmid by Lipofectamine 3000. The cells were treated with 10 μg.mL -1 Mitomycin C, 20 μM Lovastatin, and 20 mM Metformin separately. After 24 and 48 hours, the samples were collected and, protein expression was analyzed using RT-PCR, Dot blot, and ELISA., Results: A higher protein expression level was observed in treated cells 24h and 48h after treatment with all three drugs. According to real-time PCR, Metformin treatment resulted in the highest expression level within 24 h (P=0.0026), followed by Mitomycin C treatment within 48 h (P=0.0030)., Conclusion: The NS0 cell line can be regarded as a suitable host for FVIII production. FVIII protein expression level was increased by using Lovastatin, Metformin, and Mitomycin C drugs. Further investigations are suggested, and the potential application of these drugs to increase recombinant protein yield can be used to produce therapeutic proteins in the industry., Competing Interests: The authors declare no conflict of interest, (Copyright: © 2021 The Author(s); Published by Iranian Journal of Biotechnology.)

Interleukin-2 (IL-2) is a cytokine that acts in dual and paradoxical ways in the immunotherapy of cancers and autoimmune diseases. Numerous clinical trial studies have shown that the use of different doses of this cytokine in various autoimmune diseases, transplantations, and cancers has resulted in therapeutic success. However, side effects of varying severity have been observed in patients. In recent years, to prevent these side effects, IL-2 has been engineered to bind more specifically to its receptors on the cell surface, decreasing IL-2 toxicities in patients. In this review article, we focus on some recent clinical trial studies and analyze them to determine the appropriate dose of IL-2 drug with the least toxicities. In addition, we discuss the engineering performed on IL-2, which shows that engineered IL-2 increases the specificity function of IL-2 and decreases its adverse effects.

Toxoplasma gondii is an intracellular protozoan parasite that causes neuroinflammation in the brain and a constant need for peripheral leukocyte migration. Matrix metalloproteinase 9 (MMP-9) can play a major role in this neuroinflammation and be implicated in some neurological disorders, such as migraines. Therefore, the genetic polymorphism evaluation of MMP-9 in migraine patients with T. gondii infection was performed. One hundred fourteen migraine patients and 114 healthy controls were evaluated for the presence of anti-Toxoplasma IgG antibodies. Seventy-two migraine patients and 40 healthy controls were randomly selected for assessment of the MMP 9-1562C/T genetic polymorphism. In the preliminary examination, 61 (53.5%) migraine patients and 43 (37.3%) healthy controls were positive for IgG antibodies, with a significant association between T. gondii seropositivity and migraine (OR = 1.90; 95% CI = 1.21-3.223; P = 0.012). Genetic distribution for the polymorphism was not in Hardy-Weinberg equilibrium in cases but showed no significant variation in control groups (P = 0.03 and P = 0.180, respectively). A significant preponderance of the CT + TT genotype was found in migraine subjects compared to controls (P = 0.042) (OR, 1.77, CI, 1.013-2.229). The homozygote muted allele TT had a higher rate in migraine patients (6.9%). There were significant differences in CT + TT genotype between T. gondii positive and negative migraine patients (P = 0.024), but T allele frequencies had no significant variation (OR 1.7 CI, 1.084-2.44 and 0.42 CI, 0.044-3.97, respectively). In conclusion, the results may provide the first evidence for the involvement of the MMP-9 gene polymorphism in the mechanism of migraine pathology following Toxoplasma infection., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Helicobacter pylori as a common gastrointestinal (GI) pathogen must possess certain virulence characteristics to colonize the stomach, evade host immune responses, and subsequently induce GI diseases. This research aimed to investigate the expression level of two important genes, the sialic acid-binding adherence ( SabA ) and the blood group antigen-binding adhesion ( BabA ) in H. pylori strains isolated from adult patients living in the northern part of Iran, and their association with peptic ulcer disease (PUD) and gastric cancer (GC). This cross-sectional study was carried out on adult patients referring to the GI clinic of the hospitals affiliated to Babol University of Medical Sciences, Iran. New cases diagnosed with gastritis, peptic ulcer or gastric cancer were included. Endoscopic-guided gastric biopsies were examined and H. pylori positive colonies were analyzed to determine the expression of babA and sabA genes, utilizing specific primers and the SYBR Green dye. Among 175 patients with mean age of 51.6±15.6 years, 101 (57.7%) of the individuals tested positive for H. pylori infection. Statistical analysis revealed a significant correlation between sabA (P=0.003) and babA (P=0.002) gene expression and development of PUD and GC. Smoking (P=0.052), gender (P=0.004) and positive babA gene expression (P=0.009) had the greatest association with occurrence of PUD or GC in H. pylori positive patients.  In summary, the presence of the sabA gene in people infected with H. pylori increased the risk of GC compared to gastritis, while, the presence of the babA gene was significantly increased in gastric ulcer patients. Considering the diversity of H. pylori isolates and the varying results observed in different geographical regions, further comprehensive studies are required to evaluate the function of these genes in H. pylori pathogenesis and their relationship with clinical outcomes., (© The Author(s).)

Background: A combination of bioceramics and polymeric materials has attracted the research community's interest in bone tissue engineering. These composites are essential to support cell attachment, proliferation, and osteogenesis differentiation, which are vital as a classic strategy in bone tissue engineering. In this study, NiFe2O4/ZnO-coated poly L-Lactide (PLLA) was employed as a scaffold to evaluate the osteogenic differentiation capability of human adipose tissue derived mesenchymal stem cells (hAMSCs). Material and methods: The electrospun PLLA nanofibers were fabricated, coated with nanocomposite (NiFe2O4/ZnO), and evaluated by the water contact angle (WCA), tensile test, attenuated total reflectance fourier-transform infrared (ATR-FTIR) and scanning electron microscopy (SEM). Then, the osteogenic differentiation potential of hAMSCs was assessed using NiFe2O4/ZnO-coated PLLA compared to tissue culture plastic (TCP) and a simple scaffold (PLLA) in vitro conditions. Results: The adhesion, proliferation, and differentiation of hAMSCs were supported by the mechanical and biological properties of the NiFe2O4/ZnO-coated PLLA scaffold, according to SEM and 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) staining patterns. During bone differentiation, Alkaline phosphatase (ALP) enzyme activity, biomineralization, calcium content, and osteogenic gene expression (ALP, Osteonectin, Osteocalcin, Collagen type I, and Runx2) were higher on NiFe2O4/ZnO-coated PLLA scaffold than on PLLA scaffold and TCP. Conclusion: Based on our results, the osteogenic differentiation of hAMSCs on the improved biological scaffold (PLLA coated with NiFe2O4/ZnO) could accelerate due to the stimulating effect of this nanocomposite., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Shariati, Seyedjafari, Mahdavi, Maali and Ferdosi-Shahandashti.)

Background: Multidrug resistance (MDR) in Enterobacter spp. has created therapeutic challenges all over the world. The present study was conducted for evaluating the prevalence of class I integron, determining the gene cassettes and antimicrobial resistance profile of Enterobacter spp. isolates from clinical samples in Babol, North of Iran., Methods: During a 13-month period, 30 Enterobacter spp. isolates were collected from Ayatollah Rouhani Hospital, Babol, Iran. Various types of antimicrobial agents were used to determine the resistance pattern. Class I integron and associated gene cassettes were detected by PCR assay., Results: The resistance rates to AP, CPM, CTX, TM, NI, IMI, AK, CIP and GM antimicrobials were 100%, 93.3%, 33.3%, 33.3%, 30%, 20%, 20%, 20% and 13.3%, respectively. The distribution results of int genes showed that 63.3% of isolates carried the intI genes. Also, the prevalence of aadB , dfrA1 , bla OXA30 and bla PSE1 genes were estimated at 36.6%%, 33.3%, 6.6% and 0%, respectively., Conclusion: Our results showed that class I integrons have a widespread distribution among the Enterobacter spp. isolates and have clinical relevance to MDR isolates. The results confirmed the necessity for uninterrupted monitoring to prevent distribution of multidrug resistance among Enterobacter spp . strains in Iran., (Copyright © 2020, Babol University of Medical Sciences.)

In recent years, different acquired resistance mechanisms, including transposons, bacteriophages, plasmids, and integrons have been identified as involved in the spread of resistance genes in bacteria. The role of integrons as mobile genetic elements playing a central role in antibiotic resistance has been well studied and documented. Integrons are the ancient structures that mediate the evolution of bacteria by acquiring, storing, disposing, and resorting to the reading frameworks in gene cassettes. The term integron describes a large family of genetic elements, all of which are able to capture gene cassettes. Integrons were classified into three important classes based on integrase intI gene sequence. Integrons can carry and spread the antibiotic resistance genes among bacteria and are among the most significant routes of distribution of resistance genes via horizontal transfer. All integrons have three essential core features. The first feature is intI , the second one is an integron-associated recombination site, attI , and an integron-associated promoter, Pc, is the last feature. Among them, the class 1 integron is a major player in the dissemination of antibiotic resistance genes across pathogens and commensals. Various classes of integrons possessing a wide variety of gene cassettes are distributed in bacteria throughout the world. This review thus focuses on the distribution of integrons among important bacteria.

Objective: Rapid, reliable, and affordable detection of Brucella species via the molecular methods remains a challenge. In recent years, loop-mediated isothermal amplification (LAMP) is a functional nucleic acid amplification technique offering a substitute to polymerase chain reaction (PCR). So, we compared the LAMP assay with the conventional PCR for the identification of common Brucella species in Iran. In this study, LAMP assay was comprehensively evaluated against the common PCR method. A group of specific LAMP primers were used to amplify a highly specific fragment from the sequence of the Brucella abortus, bcsp31 gene. Sensitivity and specificity values of tests were done with a set of 78 (50 Brucella and 28 non-Brucella) strains., Results: A dilution series of B. abortus DNA indicated that the LAMP reaction could reliably detect 10 (fg/µl) DNA target copies per reaction within 36 min, which is 10 times greater than the PCR assay. In summary, we conclude that LAMP assay provide accurate and fast test results to identify of common Brucella species in low-complexity labs, mainly in low and lower middle income countries.

Background: Adult T-cell leukemia/lymphoma (ATLL) is a poor prognostic Hematopoietic malignancy with various therapeutic challenges, which had been classified as non-Hodgkin lymphoma. The Drug switching, as a novel, innovative and promising approach, is an opportunity to overcoming on therapeutic challenges of hard-treating disease, e.g . ATLL. Our aim is evaluating the antiproliferative and apoptotic effect of Mebendazole (MBZ) on ATLL cancer cells in in-vitro conditions. Materials and Methods: We used Jurkat cell-line as ATLL cancer cells. After treatment of MBZ in different concentrations on jurkat cells, the cell viabilities were determined by MTT assay. After IC 50 value determination, the 24-, 48- and 72-h treatments had been performed in IC 50 concentration and control to evaluating the quantitative apoptosis rate by Annexin/PI Flowcytometry and qualitative apoptosis by DAPI Nuclear staining. Also, Glucose spectrophotometry were performed to evaluate the reduced amount of glucose uptake through MBZ treatment. Results: MBZ inhibits proliferation of jurkat cells and IC 50 value had been estimated 10 μM ( P< 0.01 ). According to the flowcytometric results, increasing in drug concentration is associated with decrease cell viability and the percentage of full-apoptosis. However, it inversely correlates with percentage of early-apoptosis rate. Also, the microscopic captures of DAPI Nuclear staining confirms the flowcytometry results in qualitative manner. In addition, it was found that inhibition of glucose uptake was inversely correlated with increased MBZ concentration ( P< 0.05 ). Conclusion: MBZ potentially inhibits the proliferation of ATLL cancer cells in in-vitro condition. MBZ inhibits the growth of Jurkat cells by inducing apoptosis. Also, we suggest that indirectly inhibition of Glucose transporting occurs by MBZ, which could induce apoptosis in cancer cells., (Copyright © 2020 Tehran University of Medical Sciences.)

Background and Objectives: Clostridium difficile is responsible for 15-25% of nosocomial antibiotic associated diarrhea (AAD) cases and all cases of pseudomembranous colitis. C. difficile has two major virulence factors, toxin A (enterotoxin) and toxin B (cytotoxin). The aim of this study was to determine the frequency of C. difficile strains in patients with diarrhea in Babol' hospitals with toxigenic culture and PCR assay., Materials and Methods: One hundred stool specimens were taken from diarrheal patients in hospitals of the city of Babol. All patients had a history of antibiotic use. The samples were cultured on CCFA medium. In the next stage, toxigenic culture was performed for isolated C. difficile strains. Then, PCR assay was used to identify gdh, tcdA and tcdB genes among isolated C. difficile strains., Results: From the 100 stool samples, eight (8%) samples were positive in C. difficile culture. In toxigenic culture, two (2%) of these strains had cytopathic effects on Vero cells. All eight strains had the gdh gene. This gene is specific for C. difficile . Two strains that had cytopathic effects on toxigenic culture were positive for toxin genes., Conclusion: The frequency of toxigenic strains in different parts of the world is variable, and needs to be continually investigated. In the present study, the PCR method had a good correlation with toxigenic culture. Thus, it can replace the laborious and costly cell culture method.

Background: Helicobacter pylori (H. pylori or Hp) has been strongly associated with the peptic ulcer diseases, chronic gastritis, ulcers, and gastric cancer. Genes associated with pathogenicity have been designated for H. pylori , and some of them appear to be related to more severe clinical consequences of the infection. The present study was conducted to determine cagA, vacA, cagE, iceA1, oipA, and iceA2 genes in H. pylori strains isolated from gastroduodenal patients, who referred to Shariati hospital in Tehran, Iran. Methods: Gastric biopsy specimens were collected during endoscopy from patients, who referred to the Shariati hospital in Tehran, Iran during January and November 2015. After isolation of H. pylori from the biopsy culture, genomic DNA was extracted and subsequently used to identify H. pylori and virulence genes using specific primers. Results: The isolation rate of H. pylori strains was 65.7% (169/257). The frequency of cagA, vacA, cagE, iceA1, oipA, and iceA2 was 143 (% 84.6), 169 (100%), 131 (77.5%), 97 (57.3%), 89 (52.6%), and 72 (42.6%), respectively. Conclusion: In this study, a significant difference was observed between investigated genes and strains isolated from PUD and GC patients (p<0.05).

Background: Staphylococcus aureus (s. aureus) nasal carriers, particularly the healthcare staff can be considered as a potential source for the spread of resistant strains. The aim of this study was to determine the molecular characterization of S. aureus strains isolated among the staff nasal carriers in one of the teaching hospitals in Babol., Methods: A total of 120 nasal swabs were taken from the staff of Ayatollah Rouhani Hospital Babol during 2016. The antibiotic resistance pattern was performed by disc diffusion method for 13 antibiotics, including cefoxitin, cephalothin, teicoplanin, vancomycin, daptomycin, oxacillin, amoxicillin, amikacin, linezolid, ciprofloxacin, levofloxacin, erythromycin, rifampin, according to the CLSI 2015. Polymerase chain reaction (PCR) was used to detect mecA and pvl genes. Finally, the different SCCmec types were determined by multiplex- PCR method., Results: Among the 120 collected specimens, 40(33.3%) S. aureus isolates were approved. 28(70%) of strains were identified as methicillin-resistant Staphylococcus aureus (MRSA) and the frequency of pvl gene was confirmed 2(5%). Based on the multiplex PCR assay, four different SCCmec types were detected as 35.7% type I, 14.2% type III, 7.1% type II and 3.5% type IV. By a disc diffusion method, no resistance pattern was observed to vancomycin, while 100% of strains were resistant to amoxicillin., Conclusion: Consequently our results illustrated that isolated S. aureus strains among the staff nasal carriers via mentioned molecular characterization may lead to increase the nosocomial persistent infections in hospitalized patients and also health care workers., Competing Interests: There was no conflict of interest in this study.

Background: Enterovirus (EV) infections are one of the most common causes of aseptic meningitis in pediatrics. To diagnose EV meningitis, virus isolation in cell cultures is often time consuming and lacks sensitivity to be of clinical relevance. This makes the virus culture results difficult to interpret. The rapid detection of EVs in cerebrospinal fluid (CSF) by molecular diagnostic techniques may improve the management of patients with aseptic meningitis. The purpose of the present study was to develop a more convenient and sensitive alternative technique to viral culture. The current investigation aimed to explore the prevalence of EVs in CSF of children with suspected aseptic meningitis in northern Iran, between June 2014 and March 2015 via the one-step real-time RT-PCR technique., Methods: A single center cross-sectional study was carried out on 50 children suspected with aseptic meningitis, aged 6 months to 13 years. The presence of EV RNA in CSF samples was screened by the use of qualitative one-step real-time RT-PCR., Results: Enteroviral RNA was detected in 9 (18%) subjects using the one-step real-time RT-PCR assay. There was significant difference between EV positive and negative subjects regarding mean age (P=0.023), mean lymphocyte percentage (P=0.001) and mean glucose levels in CSF (P=0.037). The disease onset data indicate that the majority of EV meningitis occurred in the summer., Conclusion: This study provides the first data on the prevalence and epidemiology of EV infections in children with suspected aseptic meningitis in northern Iran., Competing Interests: The authors declare that they have no conflict of interest.

Background: We investigated the integron types and their relation with antibiotic resistance among A. baumannii isolates collected from intensive care unit patients, Babol, north of Iran., Methods: In this cross-sectional study, a total of 73 bronchoalveolar lavage samples were obtained from patients in ICU. Susceptibility testing was performed by disk diffusion method. Types of integrons were identified by an integrase gene PCR., Results: In total, 47.9% A. baumannii isolates were recovered from the BAL samples. All isolates were resistant to ceftazidime. 91.4% and 58.3% of isolates were MDR and XDR, respectively. The rate of colistin resistance with the E -test was 5.7%. Molecular analysis of class I, II, and III integrons showed that 25.7%, 88.6%, and 28.6% of the isolates carried the intI , intII , and intIII genes, respectively., Discussion: Our results show that different classes of integrons are commonly spread among A. baumannii strains and these genomic segments can play an important role in the acquisition of MDR and XDR phenotypes. So monitoring drug resistance in A. baumannii isolates with the use of int gene PCR is very important to plan specific infection control measures to prevent the spread of MDR-AB and XDR-AB in Iran's hospitals.

Background: Wolbachia are common intracellular bacteria that infect different groups of arthropods including mosquitoes. These bacteria modify host biology and may induce feminization, parthenogenesis, male killing and cytoplasmic incompatibility (CI). Recently Wolbachia is being nominated as a bio-agent and paratransgenic candidate to control mosquito borne diseases., Methods: Here we report the results of a survey for presence, frequency, and phylogenetic congruence of these endosymbiont bacteria in Culex pipiens populations in Northern, Central, and Southern parts of Iran using nested-PCR amplification of wsp gene., Results: Wolbachia DNA were found in 227 (87.3%) out of 260 wild-caught mosquitoes. The rate of infection in adult females ranged from 61.5% to 100%, while in males were from 80% to 100%. The Blast search and phylogenetic analysis of the wsp gene sequence revealed that the Wolbachia strain from Iranian Cx. pipiens was identical to the Wolbachia strains of supergroup B previously reported in members of the Cx. pipiens complex. They had also identical sequence homology with the Wolbachia strains from a group of distinct arthropods including lepidopteran, wasps, flies, damselfly, thrips, and mites from remote geographical areas of the world., Conclusion: It is suggested that Wolbachia strains horizontally transfer between unrelated host organisms over evolutionary time. Also results of this study indicates that Wolbachia infections were highly prevalent infecting all Cx. pipiens populations throughout the country, however further study needs to define Wolbachia inter-population reproductive incompatibility pattern and its usefulness as a bio-agent control measure.

Background: Urinary tract infection (UTI) is one of the most prevalent infectious diseases and Escherichia coli is its common cause. The aim of this study was to assess the resistance patterns of E.coli in urinary tract infections and to determine the susceptibility of E.coli to commonly used antimicrobials and also to evaluate the options for empirical treatment of UTI., Methods: This study was conducted in the Ayatollah Rouhani Teaching Hospital of Babol Medical Sciences University in North of Iran. Between January of 2013 to December 2013, antimicrobial susceptibility tests were done by disk diffusion and microdilution method. Growth of >=10(5) cfu/ml was considered as positive urine test. Ten commonly used antibiotics were examined for susceptibility test. Data and the results were collected and analyzed., Results: E.coli grew in 57 urine samples. Imipenem, ofloxacin, ciprofloxacin were the most sensitive antibiotics at 87.7%, 87.7% and 78.9% respectively. Whereas, cotrimoxazole, cefexime, cefotaxcime and ceftriaxone were the most resistant antibiotics. Antibiotic sensitivity of disk diffusion compared minimum inhibitory concentration (MIC) detected by microdilution had the sensitivity, specificity, positive predictive value and negative predictive value of 82%, 98%, 99% and 74%, respectively., Conclusion: Imipenem, ofloxacin and ciprofloxacin should be used in empirical therapy of UTI.

Background and Objectives: The rapid emergence and dissemination of carbapenemase-producing Klebsiella pneumoniae strains and other members of the Enterobacteriaceae poses a considerable threat to the care of hospitalized patients and to public health. The aim of this study was to determine the frequency of metallo-β-lactamases (MBL) and VIM-1 gene in multidrug-resistant strains of K. pneumoniae., Methods: 50 isolates of non - duplicated K. pneumoniae cultured from patients at intensive care units were tested for their susceptibilities to 13 different antibiotics using microbroth dilution assay. Isolates showing resistance to at least one of the carbapenems were checked for production of metallo-β-lactamase (MBLs) using imipenem-EDTA synergy tests. PCR was used to detect the gene encoding VIM-1 metallo-β-lactamase (MBL)., Results: Of 50 clinical isolates, 26 (52%) were resistant to imipenem in disk diffusion method. Using imipenem-EDTA synergy tests, production of MBL was detected in 15 (30%) isolates. PCR assay showed that 15 isolates were positive for VIM and these included 10 and 5 isolates showing positive and negative results in phenotypic method of MBL detection test respectively. Amikacin was found as the most effective antibiotic against the MBL producers in this study., Conclusion: The emergence of bla(VIM-1) producing K. pneumoniae in North of Iran is concerning. Microorganisms producing bla(VIM-1) constitute the prevalent multidrug-resistant population of K. pneumoniae in that region.

Background: Infection with non-typhoid Salmonella (NTS) is one of the most important health problems all over the world. Antimicrobial drug resistance is increasing among Salmonella infantis species., Objectives: The aim of this study was to investigate the frequency of presence of class 1 integrons in S. infantis species as well as its association with drug resistance., Materials and Methods: This cross-sectional study was performed on 50 S. infantis isolated strains, collected from chicken samples between 2009-2011. These strains were identified by standard biochemical tests and serology. Antibiotic susceptibility profiles and minimum inhibitory concentration determination for 14 antibacterial agents were performed using micro dilution and disk diffusion methods. The detection of class 1 integron was performed by the PCR method. The demographic and microbiological data for the integron positive and negative isolates were compared by SPSS software., Results: Eighteen out of 50 (36%) of isolated S. infantis species had intl gene. The isolated bacteria were sensitive to cefotaxime and ciprofloxacin (100%). Also isolates were resistant to nalidixic acid, tetracycline and streptomycin. All isolate with class 1 integron were multidrug resistant., Conclusions: The result of this study showed that due to increased level of drug resistance in S. infantis and the presence of class 1 integron in these strains, resistance can be transferred to other food borne pathogens.

Pseudomonas aeruginosa is one of the most important pathogens that causes nosocomial infections and shows high level of antibiotic resistance. Integrons are one of the transposable elements in bacteria and their role in antibiotic resistance has been well demonstrated. The aim of this study was a molecular characterization of the integron genes and the determination of the resistance or sensitivity pattern to ceftizoxime, cephizoxim. cephotaxim, amikacin, ofloxacin, imipenem, cefepime, ticarcillin, gentamicin, ciprofloxacin, cefazolin and ceftriaxone antibiotics in P. aeruginosa strains isolated from Intensive Care Units (ICU), Shahid Beheshti Hospital, North of Iran. This cross-sectional study was performed from 2011 to 2012. Totally, fifty four P. aeruginosa strains were isolated from ICU at Shahid-Beheshti hospital, Babol, North of Iran. The bacteria were diagnosed based on mobility, pigment production, growth in 42(0) C, oxidase and catalase tests. PCR analysis was carried out to detect integron genes using hep 35 and hep 36 primers. Also, disk diffusion method was performed to evaluate antibiotic susceptibility of the bacteria using ceftizoxime, ceftazidime, cephotaxime, amikacin, ofloxacin, imipenem, cefepime, ticarcillin, gentamicin, ciprofloxacin, cefazolin and ceftriaxone antibacterial reagents. This study revealed that 20 (37%) P. aeruginosa isolates had integron genes. The antibiotic susceptibility test showed that 53 (98.1%) of the isolates were multidrug-resistant. 12 out of 54 isolated bacteria were resistant to all antibiotics tested. All bacteria were resistant to cefepime and the highest resistance rate was seen to ceftizoxime 92.6% followed by cefazolin 92.3%. The lowest resistance rate was observed to ciprofloxacin 38.9%, ofloxacin 44.4%, amikacin 46.3% and ticarcillin 48.1%. According to this study, P. aeruginosa isolates showed high level of antibiotic resistance and the presence of integrons in these strains can explain the influence of these genes in resistance creation. There was a significant association between resistance to cefotaxime, amikacin, ofloxacin, imipenem, ticarcillin, gentamicin and the presence of integrons.