Showing 10 results for Nazarian
Volume 10, Issue 0 (بهار 86- 2008)
Abstract
Objectives: Salmonella typhimurium is important food-borne pathogen responsible for gastroenteritis. In this work a polymerase chain reaction based enzyme linked immunosorbent assay (PCR-ELISA) was developed to identify Salmonella typhimurium.
Materials and Methods: The rfb gene which is responsible for biosynthesis of the Salmonella O-antigenic lipopolysaccharide was selected as the target sequence. The selected primers amplified fragment size of 882bp of S. typhimurium. Food samples contaminated with Salmonella typhimurium as well as clinical and standard samples were used in this investigation. The PCR products randomly labeled with Dig-11-dUTP were transformed to a plate coated with streptoavidin and tested with anti digoxigenin. An internal biotin-labeled probe was used to confirm the amplified PCR product.
Results: The specificity of the assay toward S.typhimurium samples was confirmed by testing 20 Salmonella and 6 non Salmonella strains. ELISA increased the sensitivity of the conventional PCR method by approximately 1000 fold.
Conclusion: The method presented here is a rapid and specific alternative for the traditional time consuming culture methods for the detection of S. typhimurium in food and clinical samples. Due to its high specificity and sensitivity, our method finds its place as an alternative to PCR in large scale screenings, for detection of S. typhimurium.
Volume 11, Issue 0 (بهار و تابستان 87- 2008)
Abstract
Objective: Molecular epidemiology of Visceral Leishmaniasis (VL) is currently used widely for different objectives such as vector incrimination studies.
Materials and Methods: In this study three different loci including kinetoplast DNA (kDNA), ribosomal DNA (rDNA), and Cystein protease B (CPB) of Leishmania parasite genome were used for detection and identification of natural infection of sand flies of Germi district of Ardebil province, the most important VL or Kala-azar foci in Iran.
Results: The results showed that the three loci of kDNA, rDNA and CPBs are respectively more appropriate for leptomonad infection/initial screening, identification of the L.donovani complex, and discrimination of the species complex. It was also verified that both members of the complex,
L. donovani and L. infantum, are present in the study area and are transmitted to the hosts by
Volume 11, Issue 1 (Spring & Summer 2007)
Abstract
The purpose of this study is to present “differential urbanization” theory and to test this theory for the case of Iran during 50 years between 1950- 2000 for 10 statistical periods. First the differential urbanization theory has been presented and then has been pointed the case studies employed this theory. Finally, we applied this theory for analyzing urban system of Iran. The survey of this theory for Iran shows that its urban system has passed “urbanization” phase since 1950 and has passed three stage of it to 1975 respectively and since 1975 to 1995 has passed two stage of the “polarization reversal” phase and enters to “counter urbanization” phase. The results of this study shows that totally differential urbanization theory conforms to Iran’s urban system trend, but shows some differences with urban system of Iran.
Hossein Samiei Abianeh, Shahram Nazarian, Jafar Amani, Amir Sajjad Hojjati Razgi, , Mohammad Reza Rahmani,
Volume 13, Issue 2 (1-2023)
Abstract
Background and Aim: Toxins produced by Clostridium botulinum are among the deadliest compounds known that cause botulism. Currently, the detection of BoNTs in food using bioassays on laboratory mice is a very sensitive method with a detection range of 7 to 20 pg.mL-1. However, bioassay for mice is time consuming. This method is fast, highly specific, and sensitive to experiments on mice. The aim of this study was to use the modified Sandwich ELISA method to detect BoNT/B toxin.
Materials and Methods: Recombinant 370 amino acid protein was expressed from the carboxyl terminus of the binding moiety of BoNT / B toxin with a molecular weight of 45 kDa as antigen and purified by Ni-NTA affinity chromatography. IgG antibodies were isolated from mouse and rabbit sera byG protein affinity chromatography. The sensitivity and specificity of the method designed to detect recombinant BoNT/B-HcC antigen and botulinum toxin type B were evaluated.
Results: Purified rat and rabbit antibody concentrations were 3 and 4.5 mg / ml serum, respectively. The minimum concentrations of detectable protein were determined by indirect ELISA with purified mouse and rabbit antibodies at 475 and 118 pg. By optimizing the sandwich ELISA method, at least 30 ng of recombinant BoNT/B-HcC antigen and146 pg of highly specific BoNT/B were detected.
Conclusion: sandwich ELISA method can be used for accurate and sensitive identification of Clostridium botulinum toxin type B. It is necessary to evaluate the effectiveness of this method in the future to detect botulinum toxin in environmental and food samples.
Sahar Karimi, Shahram Nazarian, Fattah Sotoodehnejadnematalahi, Roohollah Dorostkar, Jafar Amani,
Volume 14, Issue 1 (FALL 2023)
Abstract
Given the global epidemic of COVID-19, it is important to design a vaccine for prevention. The virus belongs to the beta-coronavirus family and forms appendages on the surface of the glycoprotein spike virus membrane. Studies on SARS-CoV-1 and related MERS-CoV vaccines have shown that the spike protein on the virus surface is a suitable target for the vaccine. In this experimental study, we compare the recombinant fragment of spike protein (rfsp) expressed in the eukaryotic host CHO-K1 cell and prokaryotic E. coli in terms of immunogenicity, neutralizing activity, and epitopes recognition Similar to the virus strain and the ability to bind to the serum of improved patients, in two types of alpha and delta variants. The results showed that both rfSP proteins are a potential new antigen candidate for the development of the Covid 19 vaccine, but the CHO cell maintains the biological activity of the protein by performing post-translational modification processes such as glycolysis. This increases the present likelihood of developing virus-like epitopes and increases the titer of rfsp-specific antibodies in the serum of immunized mice. Therefore, priority is given to rfsp expressed in CHO cells to evaluate vaccine efficacy.
Volume 17, Issue 4 (1-2015)
Abstract
Objectives: CtxB (Cholera toxin B subunit) contributes to a vaccine's efficacy by stimulating production of the anti-CtxB antibody. Various attempts have been made to increase production of this antibody. Chitosan is a mucoadhesive polysaccharide that has tremendous potential for oral vaccine delivery in terms of its exclusive features that include biocompatibility, biodegradability, high charge density and non-toxicity. We investigated the potential for chitosan nanofibers and nanocapsules as novel carrier systems for the oral delivery of CtxB.
Methods: Antigen-containing chitosan nanofibers were prepared by electrospinning a chitosan/AcOH solution. Encapsulation of the antigen inside the chitosan nanofibers was confirmed through infrared spectroscopy analysis (FTIR). Guinea pigs were immunized with free antigen and CtxB antigen or antigen alone by direct administration of antigen-containing chitosan nanofibers into the buccal cavity. Serum immunoglobulin G (IgG) and intestinal immunoglobulin A (IgA) antibody responses were determined
Results: The results indicated that antigen in the chitosan nanofibers or nanocapsules elicited very high IgA and IgG responses. No detectable IgA and IgG responses were obtained after oral immunization with CtxB. The results of the antibody titer were analyzed using the ANOVA and LSD tests.
Conclusion: CtxB inside the nanofiber increased antibody production when administered orally. This system might be used for delivery of other antigens.
Volume 20, Issue 3 (10-2017)
Abstract
Objective: The TcpA colonization factor of pili A and the cholera toxin are the most important pathogenesis factors of Vibrio cholera that have the ability to stimulate the immune system. The aim of this study is a bioinformatics analysis of the expression of CtxB-TcpA recombinant chimeric protein in E. coli, and production of antibody against it in mice.
Methods: We designed a gene cassette that contained the CtxB and TcpA genes, and a spacer linker by using bioinformatics. Characteristics that include the structure of the chimer protein and epitopes were studied. In order to build a gene cassette, TcpA and CtxB genes were proliferated and cloned in pET28a(+). CtxB-TcpA gene expression was induced by IPTG. The produced CtxB-TcpA recombinant protein was confirmed by SDS-PAGE and Western blot analyses. Antibody produced from mice serum was isolated and confirmed by ELISA.
Results: The codon adaptation index of the optimized gene was 0.9. The prevalence ratio codons increased to 74% through codon optimization. Enzyme analysis verified the chimeric gene CtxB-TcpA cloning in the pET28a (+) expression vector. A protein with a molecular weight of 35 kDa was seen on SDS-PAGE. Its reaction with anti-histidine antibodies was confirmed by Western blot. The purified protein was 33.100 mg/l. Immunization of mice induced a serum antibody response.
Conclusion: The chimeric protein can be considered a good candidate for effective immunity against cholera.
Volume 21, Issue 1 (Spring 2018)
Abstract
Aims: Environmental pollution and exposure to toxic metals such as lead can induce to chronic and malignant diseases and has considerable complications including carcinogenicity, immunotoxicity, and neurotoxicity. The aim of this study was to compare the protective effect of clove essential oil and vitamin C on the toxicity of lead accumulation in quail eggs.
Materials and Methods: The current clinical trial study was performed on 360 quail chicks in a poultry farm of the Veterinary Medicine faculty of Shahrekord University in 2016. Quails were randomly divided into 6 groups with different diets. After intervention, at the age of 42 days, 5 eggs were gathered from each group. An Atomic Absorption Spectroscopy (AAS) was used to determine the tissue accumulation of lead in quail eggs. To measure the amount of lipid oxidation, TBARS (Thiobarbituric Acid Reactive Substances) test and Malondialdehyde (MDA) was measured. The data were analyzed using one-way analysis of variance and Tukey's post hoc test by GraphPad Prism 5 software.
Findings: The mean of lead accumulation in quail eggs in the group receiving lead and clove essential oil was significantly lower than those receiving lead (p<0.05). Also, the mean concentration of malondialdehyde in the lead intake group was significantly higher than that of the two groups receiving the lead plus the essential oil of clove or vitamin C (p<0.05).
Conclusion: The use of clove essential oil in quail diet has a more protective effect than vitamin C on the toxicity of lead accumulation in quail eggs.
Volume 21, Issue 2 (Summer 2018)
Abstract
Aims: Botulinum neurotoxins are the strongest known bacterial toxins that cause muscle paralysis due to inhibition of acetylcholine release. Design of inhibitors is still pursued as a major strategy for intracellular inhibition of poisoning caused by these toxins. Investigation of the potential function of design inhibitors, pure poison or catalytic area is essential. The aims of present study were expression and purification of recombinant catalytic domain of botulinum neurotoxin type E (BoNT/E) Type E from a synthetic gene.
Materials and Methods: In this experimental study, the sequence of the botulinum neurotoxin type E light chain was adopted from GeneBank and codons were optimized according to E.coli BL21 (DE3) codon usage. Other bioinformatics tools were exploited to reach the optimum expression of the gene in the mentioned host. The resultant (gene) was then ordered to synthesize and cloning in pET28a (+) expression vector. The recombinant vector was transferred into E. coli BL21 (DE3) host cells. The expression of the protein was induced by addition of IPTG. The expression conditions were changed to obtain a soluble expression of the protein. Then, the protein was purified by an affinity chromatography, followed by a further purification with amicon filter. SDS-PAGE was used to evaluate expression and purification of the protein and Western blotting was performed to confirm the expressed protein.
Finding: Codon Adaptation Index of the gene increased to 0.85. The third predicted structure showed good quality. The thermodynamic analysis of the mRNA structure showed that the predicted structure is stable. The soluble expression was obtained in 18°C and 18h induction by 1 mM IPTG. Protein production with higher more than 90% purity was confirmed.
Conclusion: Optimization of the protein expression conditions resulted in producing the solution in the culture medium by E. coli BL21 as host.
Volume 21, Issue 2 (Summer 2018)
Abstract
Aims: Breast cancer is the most common cancer among women around the world and in Iran and 22% of all cancers in women is included.
About 30% of cases with breast cancers are due to the proliferation or excessive expression of HER-2 protein. Chitosan is an environmentally friendly combination, due to its minimal systemic toxicity of peptide or drug delivery, the application is considered. The aims of this study were to manufactur chitosan nanoparticle containing HER-2 recombinant protein, evaluation of the properties and effect of its oral and injection immunization on spleen lymphocytes.
Materials and Methods: In this experimental study, pET-28a vector containing HER-2 gene was evaluated using PCR by universal primers. Expression of protein in E.coli was induced with IPTG. The recombinant protein was purified using affinity chromatography and evaluated by Western Blotting analysis. Chitosan nanoparticles containing recombinant protein were prepared by inonic gelation method and their size were characterized by DLS. Mice were immunized with nanoparticles and antibody titers were determined by ELISA. The response of lymphocytes in exposure to nanoparticles was evaluated by MTT assay. One way ANOVA, Duncan test and SPSS 22 software were used for statistical analysis.
Findings: The protein with a molecular weight of 18 kDa was confirmed. Yield of protein was 12mgL-1. Encapsulation efficiency of recombinant protein in nanoparticles was 70%. The average particle size was 205.2nm. Immunization of mice induced mucosal and humoral immune response.
Conclusion: Encapsulation efficiency of recombinant protein in nanoparticles is 70%. The average particle size is 205.2nm. Immunization of mice induces mucosal and humoral immune response HER-2 recombinant protein, due to the cellular and humoral immune response, is a candidate suitable for cancer diagnostic or therapeutic purposes.
