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Leptospirosis is a zoonotic disease in nearly all parts of the world, especially in tropical and subtropical areas and is prevalent in hot and humid areas. Particularly in the northern provinces of Caspian Sea in Iran by having mild and wet weather conditions, this disease is more spread to other parts of the country. Today, with evolution of the Leptospira, disease from traditional village that enhance its prevalence among farmers and fishing, modified to epidemics in urban communities that do not have good health. Thus the purpose of this review is epidemiology, prevention, diagnosis and treatment of leptospirosis. Prevalence of the disease in each zone depends on climate, geography, population, domestic and wild animals, and the abundance of surface water. Laboratory diagnosis of leptospirosis important and Micro Agglutination Test (MAT) in the diagnosis of disease is Valuable. Molecular tests such as PCR can be used in early detection of disease particularly when there are no specific antibodies. ELISA tests are also important in early screening. Disease prevention will be Possible by increasing awareness of the people at risk, regional health systems employees, and prevention from human contact with contaminated materials and vaccination of livestock and animals.

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Background and purpose: Antibody phage display library is a powerful in vitro technology for production of recombinant antibody fragments against a wide variety of antigens. However, the presence of insert-free clones in the phage libraries limited the specific enrichment of antibody fragments in many studies. The aim of this study was to protein A-aided recovery of insert-containing phages in antibody phage display library. Materials and Methods: Tomlinson antibody library was prepared according to the standard protocol. Three rounds of panning were performed in two independent groups on VERO/HER2 cells, considering a step of protein A-aided phage precipitation at the end of each round only in one group. The efficiency of this method was evaluated on the selected clones using PCR and phage-Elisa. Results: The results of PCR showed that 87% of clones contained insert after three rounds of panning using protein A-recovered phages while in the control group only 41% of clones contained insert. Also, the results of phage-ELISA showed specific enrichment of anti-HER2 antibodies in protein A-treated group. Conclusion: This method provided a simple and effective way for selective enrichment of antibody fragments in the phage display antibody libraries.

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Background and purpose: Multiple sclerosis (MS) is one the most prevalent diseases associated with neurological and non-traumatic disability in adults. This autoimmune disease affects the central nervous system (CNS) and manifests different signs and symptoms. The prevalence of disease and age of onset varies considerably across the world. This study aimed at investigating demographic, clinical characteristics, and different risk factors in patients with MS.

Materials and methods: The study was performed on 152 MS patients from May 2013-July 2014. McDonald diagnostic criteria were used for disease diagnosis. Data was recorded and analyzed in SPSS V.17.

Results: The mean age of patients was 31.6 ± 7.3 and females were three times more likely to be afflicted with MS. Most of the female patients were married. The majority of patients had benign disease (86.4%), and positive family history of MS was found in 21.2%. Plaque lesions in MRI were seen in 94% of the patients. Sensory loss, visual impairment and depression were observed in 45.7%, 43.8%, and 9.2%, respectively. Visual impairment was the first sign of the disease that was observed in most of the patients (41.3%). Fatigue and sexual dysfunction were significantly different between relapse-remitting, primary progressive, and secondary progressive form of MS (P= 0.05 and P= 0.02, respectively). Fatigue was associated with disease progression but other symptoms were not significantly different between three forms of the disease.

Conclusion: Demographic and clinical characteristics of MS in Mazandaran province were similar to those reported from other countries. MS registry is recommended to record all MS cases in order to increase knowledge on demographic and clinical characteristics, treatment options, and patient’s response to medications. Furthermore, high familial prevalence of multiple sclerosis in Mazandaran province compared with other regions in Iran calls for further genetic analyses.

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Background and purpose: Allergic asthma is a chronic inflammatory disorder with increased inflammation and bronchial over react to stimulants. Th1 and Th2 cells are the main cells involved in the pathophysiology of asthma. The function of these cells is under the influence of T-bet and GATA3 transcription factors. This study aimed to investigate the impairment of immune responses in patients with allergic asthma compared with controls.

Materials and methods: In a case-control study, 24 patients with allergic asthma and 10 healthy individuals were recruited. The Mononuclear cells (PBMCs) were isolated and cDNA was synthesized after RNA extraction. Gene’s expressions of T-bet and GATA3 were evaluated by Real-time Polymerase chain reaction and their relationships with risk factors for asthma were analyzed using statistical tests.

Results: In our study, the GATA3 gene expression in patients increased 29 times more than that in controls (P= 0.002) but the expression of T-bet declined (0.43, P =0.32). Evaluation of T-bet / GATA3 showed that this ratio was significantly lower in patients compared with that in the controls (P<0.0001).

Conclusion: Increased expression of GATA3 and significant reduction of T-bet / GATA3 ratio in patients showed disturbed immune responses in asthma. So, any remedial or control actions should focus on improving the unbalanced situation.

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Background and purpose: Increasing resistance to Quinolones in Escherichia coli and Klebsiella pneumonia in Sari, has caused many problems in treatment. Mutation in gyrA gene lead to changes in amino acids and resistance against Fluoroquinolones in E. coli and K. pneumonia. This study aimed at identifying remarkable mutations in E. coli and K. pneumonia isolates using PCR-SSCP analysis.
Materials and methods: Antibiotic sensitivity test (ciprofloxacin, nalidixic acid) was performed using Agar Disk Diffusion method. Resistance to fluoroquinolones was confirmed by E-test. (MIC experiment). We used PCR-SSCP method to detect mutation in gyrA (ser83 – asp 87) genes. Then, the PCR products were randomly sequenced.
Results: From 103 isolates, 65 (63.2 %) were E. coli and 38 (36.8%) were K. pneumoniae. In all E.coli isolates resistant to Ciprofloxacin, at least one mutation was observed. Also, in all K. pneumoniae samples resistant to Ciprofloxacin, at least one mutation was seen and in 14 samples two mutation points were observed, but in 5 samples that were sensitive to Ciprofloxacin no mutation was observed.
Conclusion: This study showed that the mutation in gyrA genes is closely related to quinolones resistance. High prevalence of quinolones resistance in E. coli and K. pneumoniae isolates requires more appropriate infection control programs.
 

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Background and purpose: Activation-induced cytidine deaminase (AID) is a B-cell specific enzyme responsible for somatic hypermutation (SHM) and class switch recombination (CSR) of antibody genes within the B-cell follicle of peripheral lymphoid organs. Ectopic overexpression of the enzyme leads to mutations in non-B cells and Escherichia coli (E.coli) genes. However, induction of mutations in E.coli by expression of AID requires highly regulated conditions. Therefore, the aim of this study was to construct a plasmid expressing AID under the control of tightly regulated temperature-sensitive promoter of bacteriophage lambda..
Materials and methods: The AID gene was isolated from PGEMT recombinant plasmid, containing AID and cloned into PTG19-T vector. Then, AID was ligated to an expression plasmid under the control of left promoter of bacteriophage lambda and mutant thermolabile cI857 repressor. Finally, the resistance marker of the engineered plasmid was replaced by tetracycline resistance gene.
Results: The whole open reading frame of AID was ligated into a plasmid containing a cI857-sensitive receptor lambda bacteriophage. The sequence and reading frame accuracy of the cloning was confirmed by electrophoresis of PCR products on agarose gel, restriction enzyme digestion, and nucleotide sequencing methods.
Conclusion: In current study, the AID was successfully cloned into an expression plasmid containing thermo-sensitive promoter of bacteriophage lambda. The recombinant plasmid could be used in different strains of E.coli to produce mutator strains.
 

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Background and purpose: The aminoglycoside modifying enzymes (AMEs) and class 1 integrons in clinical isolates of pseudomonas aeruginosa are the major factors leading to rapid spread of antibiotic resistance. The aim of the present study was to investigate the frequency of aac(6)-IIa, ant(2'')-Ia, and class 1 integrons in clinical isolates of pseudomonas aeruginosa.
Materials and methods: For this descriptive-analytic study (2017-2018), 100 clinical isolates of pseudomonas aeruginosa were collected from Sari teaching hospitals and identified by differential diagnostic tests. Antibiotic susceptibility pattern of all isolates against selected antibiotics was evaluated using standard disk agar diffusion method. Subsequently, all isolates were evaluated for the presence of aac (6) -IIa, ant (2'') -Ia genes, and class I integrons using PCR method.
Results: Out of 100 pseudomonas aeruginosa clinical isolates, 28%, 42%, and 39% were resistant to amikacin, gentamicin, and tobramycin, respectively, while 26 isolates were resistant to all three antibiotics. The results of PCR showed that 42% of the clinical isolates contained class 1 integrons. The prevalence of aac (6) -IIa and ant (2 '') - Ia genes in aminoglycoside resistant isolates was 41.8% and 13.9%, respectively.
Conclusion: Aminoglycosides are the preferred drug for the combined treatment of infections caused by Pseudomonas aeruginosa and the prevalence of resistance to them is also high, so the intermittent examination of the susceptibility pattern of these antibiotics and the genes associated with this issue is necessary to prevent the prevalence of high levels of resistance in this bacterium.
 

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 Background and purpose: Given the role of cancer stem cells in cancer, the aim of this study was to determine the expression of LGR5 marker in gastric cancer and its association with cagA genotype of Helicobacter pylori infection.
Materials and methods: A case-control study was performed in gastric biopsy specimens from antrum and body during endoscopic examination of patients attending Sari Touba Clinic, 2017-2018. After reviewing the patient records, the samples from those aged 50 and higher were studied. Case group included gastric cancer specimens with H. pylori infection (n=30) and control group included non-cancerous samples with H. pylori infection (n=30). LGR5 expression and presence of cagA were evaluated by IHC and PCR methods, respectively.
Results: The mean ages of gastric cancer and control group were 69.5±10.1 and 62.3±7.8, respectively (P= 0.003). Twenty three patients (76.7%) in cancer group and 24 patients (80%) in control group were positive for cagA genotype. Overexpression of LGR5 was observed in 15 patients (51.7%) with gastric cancer and 11 patients (39.3%) in control group (P= 0.429).  LGR5 was also overexpressed in 18 cases (40.9%) with cagA positive genotype and 8 cases (61.5%) with negative cagA genotype (P=0.22).
Conclusion: High expression of LGR5 was observed in half of patients with gastric cancer but it was not significantly associated with cagA H.pylori genotype.

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Background and purpose: Human papillomavirus (HPV) is one of the infectious agents that causes genital and non-genital warts and skin cancers in humans. The E7 protein of this virus is one of the small oncoproteins that may be the main target in therapeutic vaccines. E7 protein with HIV-1 Tat peptide (49-59), plays a protective role that cause immune Th1 and CTLs response. The aim of this study was to design a recombinant HIV-1 Tat 49-59 / HPV16, 18, 6, 11 E7 protein in vitro and the function of this antigen for MHC-1 expression.
Materials and methods: In this study, macrophage cell line J774 was used to evaluate the expression of MHC-1 and the function of Tat peptide in delivery of recombinant protein to the surface of MHC-1. J744 mouse macrophage cell line was treated by10, 50, and 100 μg of each of the proteins with and without Tat peptide. After 24 hours, the cells were collected and then analyzed by flow cytometry.
Results: In this study, J774 cells were treated and analyzed by E7-Tat and E7 proteins at different concentrations. The study showed that 10 μg of E7-Tat protein increased the expression of MHC-1, while at higher concentrations of this protein, the expression of MHC-1 molecule decreased considerably compared to the group without Tat peptide.
Conclusion: E7-Tat protein at lower concentrations can act as a stimulant for increasing the MHC-1 expression and could be used in therapeutic vaccines.

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Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has been a major concern since it first emerged. Prevention of Coronavirus 2019 disease (COVID-19) and slowing the epidemic and prevalence of the disease are the main goals of health institutions all over the world. it is very difficult to control the disease and predict its prevalence due to the continuous changes in the form of single nucleotide polymorphism (SNP) and mutations in this virus. In fact, some changes and mutations in Corona 2019 virus directly affect the pathogenicity and its transmission. The most important mutations that occur in the spike or RBD part of the SARS-CoV-2 virus, change the function of the virus, its severity of pathogenesis, and creates new variants. Mutations in these variants change the ability of the virus in binding to human cells, the rate of transmission, and make it easier to escape the immune system which plays a role in the effectiveness or ineffectiveness of vaccines. This study reviews eight major mutations in the spike or RBD region, their presence in different variants of Corona 2019 virus, and their relationship with response rate in infected people. Search keywords included COVID-19, mutation, variant, coronavirus, and respiratory infection in Google Scholar, PubMed, Springer, ScienceDirect, and Scopus databases. The main variations made from SARS-CoV-2 are Alfa (B.1.1.7), Beta (B.1.351), and Delta (B.1.617.2) that cause the infection to spread faster and the adequacy of vaccination on these variations have not been conclusively analyzed at this point.

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Background and purpose: Coronavirus disease 2019 (COVID-19) is a respiratory disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, an indirect ELISA method was designed to measure the human IgM and IgG antibodies against SARS-CoV-2.
Materials and methods: Protein sequence of nucleocapsid antigen from SARS-CoV-2 was expressed in E. coli BL21 and then was purified by chromatography. The purified protein was confirmed by SDS-PAGE and Western blotting. An indirect ELISA method was designed to measure the specific IgG and IgM antibodies against SARS-CoV-2 using recombinant N protein. The optimized ELISA method was then applied to measure the IgG and IgM antibodies in 61 infected or recovered COVID-19 patients and in 31 healthy controls. Finally, data obtained from the designed ELISA method were compared with those of a commercially approved ELISA kit.
Results: The recombinant nucleocapsid protein was successfully expressed and purified which was confirmed by SDS-PAGE and Western blotting. The amount of optical densities obtained from the designed ELISA method was similar to those of the commercial kit in 61 patients and 31 controls. The sensitivity and specificity of the designed ELISA method for IgG were 100% compared with the commercial ELISA kit, while the sensitivity and specificity for IgM were 96.72 and 96.77, respectively.
Conclusion: Serological tests alone are not suitable for diagnosis; however, their combination with molecular tests increases the accuracy and sensitivity of the COVID-19 diagnosis. These tests are also vauable for epidemiological studies.

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Background and purpose: The mature form of brain-derived neurotrophic factor (BDNF) binds to BDNF/NT-3 growth factors receptor (Trk-B). This binding leads to activation of Ras–MAPK pathway which is integrated with cell growth and proliferation. The BDNF deficiency is correlated with various diseases and affects aging and miscellaneous. In the present study we aimed to design a chimeric LAMP-BDNF protein and assess its suitability to target neuronal cells employed in silico approaches.
Materials and methods: The specific binding between the BDNF and Trk-B would be used to guide the drug loaded lysosomes in to brain tissue. So, the structure of the chimeric BDNF/LAMP2 protein was predicted and its quality was confirmed using bioinformatics tools. Then, the interaction between the structures of BDNF/LAMP2 protein and Trk-B was analyzed by protein-protein docking. The orientation of the interaction between these proteins was compared to orientation of Trk-B interaction with its specific antibody using structural superimposition.
Results: Findings showed that the chimeric BDNF/LAMP2 protein could preserve the original structure of the BDNF molecule. Moreover, the chimeric BDNF/LAMP2 protein was found to be capable of interacting with Trk-B. The interaction orientation and binding affinity for BDNF/LAMP2 protein and Trk-B were similar to the orientation and binding affinity of interaction between Trk-B and its specific antibody.
Conclusion: The designed chimeric BDNF/LAMP2b protein would be capable of targeting the exosomes toward neural cells and provides better drug delivery to brain tissue.

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In the article published in volume 32, issue 215, 2022,http://jmums.mazums.ac.ir/article-1-17695-fa.html the affilifations of Naghi Shahabi Majd, Hosein Ranjbaran, and Shabanali Khodashenas were published incorrectly, which are now corrected.
 

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Background and purpose: Pseudomonas aeruginosa is a significant contributor to hospital-acquired infections, including urinary infections, bacteremia, pneumonia, and burn wound infections. Due to the significant role of carbapenems in treating infections that are resistant to conventional antibiotics, it is necessary to identify the prevalence and resistance patterns of pathogens that cause hospital infections. This will help to prevent the spread of microbial resistance, and subsequently, reduce the death rate caused by such infections. Therefore, the aim of the present study is to determine the occurrence of metallo-beta-lactamase genes in Pseudomonas aeruginosa isolates in Sari hospitals.

Materials and methods: This cross-sectional study included 150 isolates of Pseudomonas aeruginosa as clinical samples that were collected from patients from the burn department of Zare Hospital, Sari city. To determine the antibiotic sensitivity, the disk diffusion method was used. Additionally, PCR method was employed to examine the prevalence of beta-lactamase genes blaVIM-1 and blaIMP-1 in the isolates.
Results: Based on the antibiotic sensitivity results, it was found that the highest resistance was observed to gentamicin, ceftazidime, imipenem, ciprofloxacin, amikacin, meropenem, cefepime, ceftriaxone, and calcitin. The data analysis also revealed that the frequencies of blaVIM-1 and bla IMP-1 genes were detected in isolates of 45% and 55%, respectively.
Conclusion: The frequency of Pseudomonas aeruginosa strains carrying the MBLs genes is increasing in Zare Hospital in Sari. The findings of the study strongly suggest that there is necessary to prevent the experimental and indiscriminate administration of carbapenems in intensive care and burn departments of hospitals.
 

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Background and purpose: Breast cancer is one of the most common cancers among women. Abnormal expression of microRNAs is associated with cancer. Therefore, this study aimed to evaluate the expression of miRNA-210 in the serum of breast cancer patients.
Materials and methods: The studied population included 49 breast cancer patients and 55 healthy individuals, and the samples were evaluated using Real Time PCR.
Results: The data analysis of this study revealed that the expression level of miR-210 in the blood of breast cancer patients is significantly higher compared to the blood of healthy people. Also, there is a negative correlation between the age of breast cancer patients and the expression of miR-210 (r=-0.309, P=0.031) and a significant positive correlation between the increase in the expression of mir-210 and the expression of the Ki-67 marker (r=0.412, P = 0.004).
 

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Background and purpose: Chronic Lymphocytic Leukemia (CLL) is a lymphoproliferative disorder characterized by the clonal and malignant proliferation of mature B-cells within the peripheral blood, bone marrow, and secondary lymphoid tissues. In chronic infections and cancers, T-cells become exhausted due to continuous antigen exposure. This uncontrolled tumor cell growth creates an environment that lacks sufficient oxygen, nutrients, and physical space for normal cells, referred to as the tumor microenvironment. The tumor microenvironment employs multiple mechanisms to suppress the immune system, affecting metabolic pathways in both T-cells and tumor cells. Consequently, inhibitory receptors on T-cells and their ligands on tumor cells are upregulated, leading to reduced proliferation, cytotoxicity, and production of effector cytokines, such as interferon-gamma, by T-cells. T-cell exhaustion presents a therapeutic challenge in treating many malignancies, including CLL. In CLL, T-cell exhaustion is observed through the simultaneous upregulation of inhibitory receptors. Given that current therapies for CLL are often associated with side effects, resistance, and relapse, studying transcript changes may assist in designing more effective treatment strategies. This study investigates the transcriptome profile of CD8⁺ T-cells in CLL patients compared to healthy individuals using bioinformatics platforms.
Materials and methods: The keywords used in this study were CLL, CD8, and human. Raw data were initially searched in the SRA database using these keywords. Left and right reads were merged using CLC Genomics Workbench version 21. Following quality control of the data, sequences were assembled using human genome reference data, mRNA sequences, and coding sequences (CDS). Gene expression related to CD8+ T-cell exhaustion was then compared between CLL patients and healthy individuals. The analysis results were presented in heatmap and volcano plot formats.
Results: In this study, genes involved in CD8⁺ T-cell exhaustion and differential expression were identified by comparing 11 CLL patients with 11 healthy individuals. The expression levels of 59 selected genes were displayed in volcano plot and heatmap formats. Expression changes in 42 genes were statistically significant (FDR P<0.05), with 18 genes showing increased expression and 24 showing decreased expression in CLL patients compared to healthy controls.
Conclusion: The study found differences in the expression levels of genes involved in CD8⁺ T-cell exhaustion between CLL patients and healthy individuals. Molecules encoded by genes upregulated in CLL may serve as potential targets for developing new therapies. By creating monoclonal antibodies and small molecules to inhibit these targets, it may be possible to counteract T-cell exhaustion, representing a promising advance in the treatment of this phenomenon.