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Copyright of Archives of Razi Institute is the property of Institut Razi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Summary: Several explanations have been suggested concerning the variety in bacille Calmette–Guerin (BCG) vaccine efficacy on strains of Mycobacterium tuberculosis (Mtb). This study aimed to compare the effect of BCG vaccination history in the prevention of the occurrence of Mtb‐Beijing and non‐Beijing strains. In this cross‐sectional study, 64 patients with pulmonary tuberculosis (TB) were recruited from the Iranian border provinces (North West and West). Isolates were subjected to restriction fragment length polymorphism (RFLP) analysis, using the insertion sequence IS6110 as a probe (IS6110 RFLP) and drug susceptibility testing using the proportion method. Samples were analyzed with Gel Compare II 6.6 and spss version 18. The mean age [standard deviation (SD)] of the patients was 54·4 (SD = 17·0). Overall, 49 cases (76·56%) had no BCG vaccination scar. The prevalence of Beijing strains was 9·38% and drug resistance proportion among the isolates was 14·1% (nine cases). There was a significant relationship between Beijing strains and tuberculosis (TB)‐drug resistance in isolates (χ2 = 26·29, P < 0·001). There was also a strong association between vaccination history and Beijing strains (χ2 = 13·23, P = 0·002). Also, a statistical relationship was observed between Beijing strains and drug‐resistant TB among patients with a history of vaccination (χ2 = 7·47, P = 0·002). This association was not maintained in the unvaccinated group (P = 0·102). These findings confirm the claim that the vaccine has different effects on different subspecies of tuberculosis. The cause of the high probability of drug resistance in patients with Beijing‐TB and vaccination history requires further investigation with a higher sample size. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis ( VL) is caused by Leishmania infantum in the Mediterranean basin and affects primarily children and immunosuppressed individuals. Various strategies of vaccination have so far been examined by either protein or DNA without achievable complete protection against the disease. The live non-pathogenic lizard parasite, Leishmania tarentolae, expressing elected Leishmania antigens has recently provided a promising new approach as a safe and effective live vaccine candidate to prevent leishmaniasis. Here, we evaluated the immunoprotective potential of a live recombinant L. tarentolae expressing Lipophosphoglycan 3 ( LPG3) antigen against L. infantum infection in BALB/c mice. Results indicated that the administration of live recombinant Leishmania produced a significant high level of IFN- γ accompanied by reduced levels of IL-10 as compared to wild-type parasites as live vaccine control, thus suggesting the induction of a Th1-type immune response in a mouse model of visceral leishmaniasis. Analysis of the IgG antibody response also showed high levels of IgG2a relative to IgG1 in sera of mice immunized with recombinant Leishmania parasites. However, immune responses elicited by this live vaccine conferred partial protection against infectious challenge. Therefore, further studies are required to increase its protective efficacy. [ABSTRACT FROM AUTHOR]

Copyright of Archives of Razi Institute is the property of Institut Razi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Identifying Mycobacterium tuberculosis (MTB) transmission type is a key step in the control of this disease. Aim: This study aimed to determine the path and transmission type of MTB and the insertion sequence IS6110 band number and verify their relationship to demographic and clinical risk factors. Subjects and Methods: In this cross-sectional study, 64 MTB patients from three border provinces of Iran were selected after full clinical history and physical evaluation design. The drug susceptibility testing was carried out using the standard proportion technique on sputum samples. Isolates tested with restriction fragment length polymorphism technique used IS6110. Results: Recent transmission of disease was 33/50 (66%) based on clustering rate. The IS6110 band number had a significant relationship with drug resistance detected in proportion method tested by univariate linear regression (P < 0.01). Furthermore, the IS6110 band number had association with Bacillus Calmette-Guérin vaccination history (P = 0.02), sex (P < 0.01), and purified protein derivative (PPD) reaction size (P < 0.01) tested by multiple analysis. The risk of recent transmission inferred from the clustering rate was significantly higher in patients from Western provinces compared to those from the North-West province (P = 0.048). However, age (P = 0.39), gender (P = 0.16), vaccination history (P = 0.57), drug susceptibility, and PPD (P = 0.6) were independent of clustering. The largest cluster of up to six subjects was found in the Western provinces. Conclusion: Recent MTB transmission was much more common in the West compared to the North-West of Iran. Large MTB clusters with strong epidemiological links may be reflective of a disease outbreak. Correlation noted between the IS6110 band number and vaccination history; PPD size and female gender necessitates further studies. [ABSTRACT FROM AUTHOR]

Background and Objectives: Rituximab is an anti-CD20 chimeric monoclonal antibody widely used for the treatment of malignant B cells lymphoma. However, the immunogenicity of murine-derived monoclonal antibodies and the large size of full length antibodies restrict cancer immunotherapy. Humanized single chain antibodies can be a solution and a promising alternative for application in immunotherapy. The aim of this study was to produce a humanized scFv antibody for a potential use in the diagnosis and treatment of B cell lymphoma. Materials and Methods: We used a CDR grafting based approach to design a humanized scFv gene fragment. The CDRs were grafted onto the closest human frameworks. The designed sequence was expressed in E.coli then purified. The level of expression was analyzed by SDS-PAGE and the reactivity to CD20 expressing cell line was explored by immunoblotting. Results: Similarity analyses revealed that human germline gene IGHV1-46*03 and IGKV1-39*01 have the highest homology with their murine counterparts. Analysis by SDS-PAGE exhibited a high expression level in E. coli. Reactivity to CD20 expressing Raji cells showed that the produced antibody maintained the binding capacity to human CD20 marker. Conclusion: In our study, humanized anti- CD20 scFv indicated an original antigen-binding affinity. The findings serve as a basis for the development of novel therapeutic strategies in the treatment of CD20- expressing cancers. [ABSTRACT FROM AUTHOR]

The main objective of this study was to investigate the effect of enterococci isolated from traditional Lighvan cheese on the quality of Iranian UF white during ripening. Four samples of cheese were provided from four different cheese production units in Lighvan region. Strains of enterococci in these samples were isolated by standard microbiological methods and selective medium of Kanamycin Esclin Azide Agar and then identified by biochemical methods. In the second stage of research, the effect of adding isolated enterococci in traditional Lighvan cheese on the quality of Iranian UF white cheese was investigated in a 60-day period. Addition of Enterococcus spp. did not significantly (P> 0.01) affect the pH and percentage of pH 4.6-Soluble nitrogen/total nitrogen. In the cheese produced with E. faecalis and E. faecium strains, lipolysis rate was higher and flavor properties were improved. Moreover, results of measuring percentage of soluble nitrogen at pH 4.6 and urea polyacrylamide gel electrophoresis indicated an increase in proteolysis rate in the cheese containing E. faecalis and E. faecium strains compared to the control cheese. Furthermore, the highest percentage of non- protein nitrogen was observed in the cheese containing E. faecium. In conclusion, the results showed the positive effect of the E. faecalis and E. faecium on secondary proteolysis during ripening. The proteolytic activity displayed by some enterococcal strains may contribute to cheese ripening and flavor development. Because of these interesting metabolic traits, enterococci have been proposed as part of defined starter culture combination for UF white cheeses. [ABSTRACT FROM AUTHOR]

Background: Genome - wide analysis of genetic predisposition to type 2 diabetes mellitus in different populations have yielded variable results. Calpain10, a gene that encodes a non-lysosomal cysteine protease, has been recently proposed as a type 2 diabetes susceptibility genes in the non-insulin-dependent diabetes mellitus NIDDM1 region. Aim of the study is to evaluate the relation between SNP43 and type2 diabetes. Methods: A total of 102 diabetic subjects and 100 undiabetic controls enrolled in a case-control study in Eastern Azerbaijan Azerbaijan Province. The polymerase chain reaction restriction fragment length polymorphism technique (PCR-RFLP) was applied. We use X² test and logistic regression to analysis of data. Results: Genotypes distribution of calpain10 gene in control group were 11(11%), 86(86%) and 3(3%) with respected to A/G, G/G and A/A genotypes, respectively. In diabetic group genotypes distribution were 7(6.9%), 95(93.1%) and zero, respectively. The G allele frequency was significant difference in case and control groups. Conclusion: Since G allele is a risk factor to affect type 2 diabetes disease; hence SNP43 of calpain10 gene had significant association with type 2 diabetes in Eastern Azerbaijan. [ABSTRACT FROM AUTHOR]

Embryo transfer is a reproductive technique that has a major impact on the dissemination of economically important genes and the rate of genetic gain in breeding schemes. In recent years, there has been increasing interest in the use of sexed and genotyped embryos in commercial embryo transfer programs. Marker/gene assisted selection (MAS / GAS) projects can be performed in the pre-implantation stage through mass production of characterized embryos. Biopsy of a few cells in the morulla stage is essential for pre-implantation genetic diagnosis (PGD), in which sex determination, evaluation of disease genes, and genotyping for candidate genes are performed. Limited quantity of cells and low amount of DNA restrict the use of multiple molecular analyses in PGD programs. Recently, whole genome amplification (WGA) techniques promise to overcome this problem by providing sufficient input DNA for analysis. Among several techniques proposed for WGA, the primer extension pre-amplification (PEP) and the improved-primer extension pre-amplification (I-PEP) methods are the most commonly used. However, these methods are time-consuming and need more than 12 h amplification cycles. Since the time is a critical parameter in the successful characterized embryo transfer, the shortening of diagnosis time is highly desirable. In this study, we developed a short and simple I-PEP procedure (∼3 h) and evaluated its performance for the amplification of bovine genomic DNA. We assessed short WGA procedure by polymerase chain reaction (PCR) amplification of 7 specific loci. The results indicated that the short procedure possesses enough sensitivity for the molecular genetic analysis of 1 input cell. Although the efficiency of the method was 100%, there was an inconsistency between genomic DNA (gDNA) and whole genome amplification product (wgaDNA) genotypes for kappa-casein locus; that is, however, most likely due to allele drop-out (ADO) or false homozigocity. The results of this study indicate that with the application of reliable methods, WGA-amplified bovine DNA will be a useful source for sexing and genotyping bovine embryos in several quantitative trait locus (QTL) markers. [ABSTRACT FROM AUTHOR]

Rapid and sensitive detection techniques for indicator pathogens are important in pharmaceutical industry. However, common detection methods rely on bacterial culture in combination with biochemical tests, a process that typically takes 5–6 days to complete. Thus, the aim of this study was to develop a multiplex polymerase chain reaction ( mPCR) assay for simultaneous detection and identification of four indicator pathogenic bacteria in a single reaction. Specific primers for indicator bacteria, namely Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella, were applied to allow simultaneous detection of them, and the sensitivity and specificity of each primer pairs were determined. In the mPCR with mixed DNA samples, specific bands for corresponding bacteria were simultaneously detected. Agarose gel electrophoresis of PCR products revealed 100% specificity of mPCR with single bands in the expected sizes. Low levels of microbial contamination less than 10 cfu per milliliter or gram of product were detected using mPCR assay. The detection of all four indicator pathogenic bacteria were completed in less than 8 h with this novel mPCR method, whereas the conventional United States Pharmacopeia methods and uniplex PCR required 5–6 days and 27 h for completion, respectively. Using mPCR assay, the microbial quality control of nonsterile pharmaceutical products can be performed in a cost-effective and timely manner in pharmaceutical industry. PRACTICAL APPLICATIONS Detection of pathogenic indicatiors of Escherichia coli, Staphylococcus aureus, Salmonella and Pseudomonas aeruginosa is one of the mandatory tests in microbial quality of nonsterile pharmaceutical products; therefore, rapid and sensitive detection of the contaminations is of great importance for product release. According to the results of the present study, simultaneous detection of low levels of four major potential pathogenic bacteria in pharmaceutical finished products can be performed using mPCR in a cost-effective and timely manner, and upon these properties of the mPCR assay it could have potential applications in pharmaceutical industry. [ABSTRACT FROM AUTHOR]

TheLeishmania majoramastigote class I nuclease (LmaCIN) is a developmentally regulated protein that is highly expressed in the amastigote stage ofL. major. This protein is homologous to the P4 nuclease ofL. pifanoi,which has been shown to induce protective immune response in a murine model. To evaluate LmaCIN as a potential human vaccine candidate, cellular immune responses to recombinant LmaCIN were examined in individuals recovered from Old World cutaneous leishmaniasis. Peripheral blood mononuclear cells (PBMC) from patients recovered fromL. majorinfection were cultured either with recombinant LmaCIN or autoclavedL. major(ALM) as control. rLmaCIN induced significant proliferation of PBMC from 90% of recovered patients. Phenotypic analysis of proliferating cells showed that CD8+ cells were the predominant cell type proliferating in response to rLmaC1N. Screening of culture supernatants for cytokines showed that rLmaCIN induced high levels of interferon (IFN)-γ (mean± s.e.m.: 1398 ± 179 pg/ml) associated with little interleukin (IL)-10 and little or no IL-5 production. These findings show that LmaCIN is immunogenic in humans duringL. majorinfection and that it can elicit immunological responses relevant to immunoprophylaxis of leishmaniasis. [ABSTRACT FROM AUTHOR]