Frequency of Prevalence of Klebsiella pneumoniae in Clinical Samples and the Evaluation of the Role of Efflux Pump in Determining Antibiotic Resistance

In this study 193 Klebsiella pneumoniae were isolated from urinary tract infections, ulcers, sputum and blood. Initially, Mac agar medium was used to isolate the bacterium, and for each suspected isolate, pink and aqueous colonies were stained and biochemical tests of catalase, oxidase, TSI, IMVIC Test and urase were performed. Confirmation of the isolates using 16SrRNA sequencing Some isolates are evaluated. Then all isolates evaluated for sensitivity to antibiotics such as Ampicillin, Amoxicillin clavulanate, Piperacillin, Cefoxitin, Cefuroxime Imipenem, Tetracyclines, Nitrofurantoin, Polymyxin B Colistin, they use disk diffusion test. In the process, the presence of the acr efflux pump gene is confirmed by using an specific primer namely Acr primer, and finally, using phenylalaninearginine- beta naphthylamide inhibitor, the relationship between antibiotic resistance and efflux pump function is evaluated. Overall, 50.2% of the collected samples contained Klebsiella. Thus, 193 of 384 clinical specimens contained Klebsiella. Of the 193 positive samples, the groin lesions had the highest percentage and the abscess had lowest percentage of Klebsiella infection, although Klebsiella was significantly separated from the throat, sputum, catheter and foley. Antibiotics, cefazolin, ceftazidime, ciprofloxacin, Chloramphenicol, tetracycline had higher antibacterial activity. Results were analyzed by Whonet 5 and SPSS software

Design and Production of Recombinant TAT Protein Structure, Catalytic Domain of Diphtheria Toxin, and Evaluation of Its Effect on Cell Line

Background and Objectives: Cancer is one of the most deadly diseases in the present age and its conventional therapies have had low success. Toxin therapy of cancer is a new therapeutic approach, which has attracted the attention of pharmaceutical specialists. Diphtheria toxin consists of three functional, transducing, and binding domains, that the functional part inhibits protein synthesis and causes cell death. The purpose of this study was to produce the functional domain of the diphtheria toxin fused with a TAT penetrating peptide and to investigate its degree of lethality on cell line.   Methods: In this study, primer was designed for tat-diph gene and the gene construct containing the sequence of the functional chain of diphtheria toxin and the TAT peptide sequence, were amplified and cloned into prokaryotic expression vector pET-28a containing a sequence of histidine. After transferring into bacterial host (E. coli BL21 DE3), induction of expression was performed using IPTG, then, the resulting protein was purified and the protein expression was confirmed by anti-histidine antibody attached to horseradish peroxidase (HRP) using western blotting technique. In the next step, the lethality of the produced chimeric protein, was evaluated on MCF7 cell line by MTT assay. Data were analyzed by uni pirate and oligo 6 software.   Results: The results of MTT assay indicated that the TAT-Diph recombinant protein adjacent to the MCF7 cell line increased the rate of mortality of the cells.   Conclusion: The results of this study revealed that the recombinant protein with functional domain of diphtheria toxin and AT peptide could have lethal effect on MCF7 cell line

Formulation of hepatitis B vaccine in MF59 adjuvant and comparison of its immunization with Iran\'s commercial hepatitis B vaccine

Introduction: Hepatitis is a systemic diseasethat causes liver inflammation. The prevention of this infection is a vaccination. The commonly used vaccine to fight this disease is to use the vaccine formulated with Alum. This vaccine cannot provide immune response and complete productivity in some people. In this study, cellular and humoral immune responses of hepatitis B vaccine were compared with hepatitis B vaccine formulated in MF59 adjuvant. Methods:  In this experimental study, Balb/c mice received different formulations of the vaccine subcutaneously three times with a two-week interval. Then, the mice were bled and the levels of anti-HBs Ag were determined by the ELISA method. IFN-γ, IL-4, IL-2 and IFN-γ / IL-4 cytokines were examined by the ELISA method from the soup of spleen cells culture. The data were analyzed using the GraphPad prism software ANOVA. Results: IL-4 levels were significantly higher in alum vaccine than the vaccine formulated in MF59, also the IFN-γ cytokine level showed no significant difference between two main groups. TNF-α cytokine shows that alum vaccine is more secreted due to the high inflammation compared with the vaccine with MF59. Total antibody in the third injection, in some dilutions of the commercial vaccine was more than vaccine with MF59. Conclusion: Significant decrease in IL-4 and antibodies indicates that the tendency of vaccine formulated in MF59 to induce cellular immune responses is higher than humoral immune responses. In addition, the safety and lack of side effects of the MF59 adjuvant can also be considered as another advantage