Design and Production of a Novel Recombinant Chimeric IL2-Omp31 Antigen against Brucella Infection

Brucellosis is a zoonotic disease in human and animals. Brucella melitensis is one of the most pathogenic species of Brucella in goat and sheep. Omp31 is an outer membrane protein of Brucella that acts as an immunogenic protein. Cytokines are glycoproteins with low molecular weight that play the role of an immune adjuvant and regulate immune responses. Interleukin-2 is one of the most important cytokines, which are secreted by the white blood cells and involved in T cell immune responses. In the present study, a chimeric Omp31-Interleukin2 recombinant protein was generated by means of genetic engineering techniques. This chimeric coding sequence was amplified by using specific primers and using Splicing Overlap Extension (SOE) PCR technique. The fusion of the two mentioned proteins was accomplished using a rigid linker. The generated chimeric IL2-Omp31 fragment was TA cloned, and then subcloned into pEt22b vector as an expression vector. The chimeric protein was successfully expressed in E. coli BL21 (DE3) and confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and also Western-blotting analysis. Finally, in order to assess the antigenic features of the recombinant chimeric IL2-Opm31 protein, its secondary structure and antigenicity were predicted in silico

Relationship between urbanization and cancer incidence in Iran using quantile regression

Quantile regression is an efficient method for predicting and estimating the relationship between explanatory variables and percentile points of the response distribution, particularly for extreme percentiles of the distribution. To study the relationship between urbanization and cancer morbidity, we here applied quantile regression. This cross-sectional study was conducted for 9 cancers in 345 cities in 2007 in Iran. Data were obtained from the Ministry of Health and Medical Education and the relationship between urbanization and cancer morbidity was investigated using quantile regression and least square regression. Fitting models were compared using AIC criteria. R (3.0.1) software and the Quantreg package were used for statistical analysis. With the quantile regression model all percentiles for breast, colorectal, prostate, lung and pancreas cancers demonstrated increasing incidence rate with urbanization. The maximum increase for breast cancer was in the 90th percentile (β=0.13, p-value < 0.001), for colorectal cancer was in the 75th percentile (β=0.048, p-value < 0.001), for prostate cancer the 95th percentile (β=0.55, p-value < 0.001), for lung cancer was in 95th percentile (β=0.52, p-value=0.006), for pancreas cancer was in 10th percentile (β=0.011, p-value < 0.001). For gastric, esophageal and skin cancers, with increasing urbanization, the incidence rate was decreased. The maximum decrease for gastric cancer was in the 90th percentile(β=0.003, p-value < 0.001), for esophageal cancer the 95th (β=0.04, p-value=0.4) and for skin cancer also the 95th (β=0.145, p-value=0.071). The AIC showed that for upper percentiles, the fitting of quantile regression was better than least square regression. According to the results of this study, the significant impact of urbanization on cancer morbidity requirs more effort and planning by policymakers and administrators in order to reduce risk factors such as pollution in urban areas and ensure proper nutrition recommendations are made. © 2016, Asian Pacific Journal of Cancer Prevention

Evolution of vertebral numbers in primates, with a focus on hominoids and the last common ancestor of hominins and panins

The primate vertebral column has been extensively studied, with a particular focus on hominoid primates and the last common ancestor of humans and chimpanzees. The number of vertebrae in hominoids—up to and including the last common ancestor of humans and chimpanzees—is subject to considerable debate. However, few formal ancestral state reconstructions exist, and none include a broad sample of primates or account for the correlated evolution of the vertebral column. Here, we conduct an ancestral state reconstruction using a model of evolution that accounts for both homeotic (changes of one type of vertebra to another) and meristic (addition or loss of a vertebra) changes. Our results suggest that ancestral primates were characterized by 29 precaudal vertebrae, with the most common formula being seven cervical, 13 thoracic, six lumbar, and three sacral vertebrae. Extant hominoids evolved tail loss and a reduced lumbar column via sacralization (homeotic transition at the last lumbar vertebra). Our results also indicate that the ancestral hylobatid had seven cervical, 13 thoracic, five lumbar, and four sacral vertebrae, and the ancestral hominid had seven cervical, 13 thoracic, four lumbar, and five sacral vertebrae. The last common ancestor of humans and chimpanzees likely either retained this ancestral hominid formula or was characterized by an additional sacral vertebra, possibly acquired through a homeotic shift at the sacrococcygeal border. Our results support the ‘short-back’ model of hominin vertebral evolution, which postulates that hominins evolved from an ancestor with an African ape–like numerical composition of the vertebral column

Safety and effectiveness of high-dose vitamin C in patients with COVID-19: a randomized open-label clinical trial

Background: Vitamin C is an essential water-soluble nutrient that functions as a key antioxidant and has been proven to be effective for boosting immunity. In this study, we aimed to assess the efficacy of adding high-dose intravenous vitamin C (HDIVC) to the regimens for patients with severe COVID-19 disease. Methods: An open-label, randomized, and controlled trial was conducted on patients with severe COVID-19 infection. The case and control treatment groups each consisted of 30 patients. The control group received lopinavir/ritonavir and hydroxychloroquine and the case group received HDIVC (6 g daily) added to the same regimen. Results: There were no statistically significant differences between two groups with respect to age and gender, laboratory results, and underlying diseases. The mean body temperature was significantly lower in the case group on the 3rd day of hospitalization (p = 0.001). Peripheral capillary oxygen saturations (SpO2) measured at the 3rd day of hospitalization was also higher in the case group receiving HDIVC (p = 0.014). The median length of hospitalization in the case group was significantly longer than the control group (8.5 days vs. 6.5 days) (p = 0.028). There was no significant difference in SpO2 levels at discharge time, the length of intensive care unit (ICU) stay, and mortality between the two groups. Conclusions: We did not find significantly better outcomes in the group who were treated with HDIVC in addition to the main treatment regimen at discharge. Trial registration irct.ir (IRCT20200411047025N1), April 14, 2020 © 2021, The Author(s)

Cloning and molecular characterization of Omp31 gene from Brucella melitensis Rev 1 strain

Brucellosis, caused by the genus Brucella bacterium, is a well-known infection among domestic animals. Considering the serious economic and medical consequences of this infection, various preventive efforts have been made through using recombinant vaccines, based on outer membrane protein (OMP) antigens of Brucella species. The objective of the present study was to clone, analyze the sequence, and predict the epitopes of Omp31 gene as a major B. melitensis antigen. The full-length open reading frame (ORF) for this gene was amplified by specific primers and cloned into the pTZ57R/T vector. The gene sequence of B. melitensis Rev 1 strain was submitted to NCBI database. The results of phylogenetic analysis showed that Omp31 is almost similar in different Brucella species. Online prediction software programs were also used to predict B- and Tcell epitopes, secondary and tertiary structures, antigenicity, and enzymatic degradation sites. The bioinformatic tools in the current study were confirmed by the results of three different experimental epitope prediction studies. Bioinformatic analysis identified one T-cell and three B-cell epitopes for Omp31 antigen. Finally, based on the antigenicity and proteosome recognition sites, common B- and T-cell epitopes were predicted for Omp31 (amino acids 191-204). Bioinformatic analysis showed that these regions had proper epitope characterization and could be useful for recombinant vaccine development

Genotypic characterization by multi locus variable number of tandem repeats analysis international Bordetella pertussis vaccine strains

Background: In 1930's first whole cell pertussis vaccines became available to the public heralding a dramatic success in overcoming the global burden of the disease. To date only a handful of B. pertussis strains have been used by international/local pertussis vaccine manufacturers. Inevitable well-documented genetic changes in the world population of this pathogen have prompted serious questions on suitability of traditional vaccine strains protect human against currently circulating wild isolates of Bordetella pertussis. Objective: Analyzing the genetic diversity within the most frequently-used vaccine strains of B. pertussis in the world Methods: A recently developed multi locus variable number of tandem repeats analysis (MLVA) genotyping system along with a bioinforamtic piece of analysis was conducted on 11 strain/sub-strains of B137, B203 (10536), C393, Cs, E476, Tohama I, J445 (134), B202 and J446 (509) plus 2 sub-strains of 134 and 509 that are used at Razi institute for preparation of pertussis vaccine. In this study have used 6 individual loci of VNTR1, VNTR3a, VNTR3b, VNTR4, VNTR5 and VNTR6. Findings: Six distinct genotypes were recognized among the examined strains by comparing our data with the Dutch MLVA databank. These were all new and not reported before in the database. Conclusion: This observation reiterates on necessity for detection of predominant native strains to include in vaccine preparations suitable for different countries. Keywords: Pertussis, Strain, Vaccine, Genotypin

Prioritizing methods of control and reduce noise pollution in Larestan cement Factory using analytical hierarchy process (AHP)

Introduction: exposure to noise pollution leaves Different effects on human. Goal in this Paper is the prioritizing methods of control and reduce noise pollution in Larestan cement Factory using analytical hierarchy process (AHP). Methods: For screening criteria and methods used in AHP technique, Delphi method was used. After polling of 15 experts, 8 criteria and 9 methods was selected from their consensus. Then, in order to prioritizing methods of reduce and control noise pollution, carried out Paired comparison of the methods and criteria, by experts using Analytical Hierarchy Process. Results: result of paired comparison of criteria show that initial investment cost was the most important criteria with the relative weight of 0.247, satisfaction from using of method, Account for the least important with the relative weight of 0.035. A paired comparison of methods according to the target of selecting control methods show that Personal exposure to noise Control method with the weight of 0.224 .was the first priority, and Insulation of building’s Method with the weight of 0.067 was the last priority. Conclusion: Because of personal exposure to noise control method obtained as the best method of controlling noise pollution in this Factory, ACGIH instruction about the time of noise exposure in the workplace suggested to directors

In silico analysis of Omp25 and BLS Brucella melitensis antigens for designing subunit vaccine

Brucellosis is a well-known infection in domestic animals which caused by Brucella bacterium. Due to serious economic and medical consequences of this disease, various efforts have been made to prevent the infection through the use of recombinant vaccines based on Brucella outer membrane protein (OMP) antigens. The objectives of the present study were cloning, sequencing and epitope prediction of Omp25 and BLS genes as two major Brucella melitensis antigens. The full-length open reading frame (ORF) of Omp25 and BLS genes were amplified and cloned into pTZ57R/T vector. Phylogenetic analysis of sequenced genes showed that both genes were nearly similar in different Brucella species. Several online prediction softwares were used to predict B and T-cells epitopes, secondary and tertiary structures, antigenicity ability and enzymatic degradation sites. Bioinformatic tools used in the current study were confirmed by the results of three different experimental epitope predictions. Bioinformatic analysis identified five and two B-cell and two and one T-cell epitopes for Omp25 and BLS antigens, respectively. Finally, according to the antigenicity ability and proteosomal recognition site common B and T-cell epitope was predicted for Omp25 (154-162 amino acids) and BLS (37-48 and 119-139 amino acids). Results of this study might be useful for recombinant vaccine development

The examination of relationship between socioeconomic factors and number of tuberculosis using quantile regression model for count data in Iran 2010-2011

Background: Poverty and low socioeconomic status are the most important reasons of increasing the global burden of tuberculosis, not only in developing countries but also in developed countries for particular groups. The purpose of this study was to assess the association between socioeconomic factors and the number of tuberculosis patients using quantile regression for count data. Methods: This cross-sectional study was conducted on 11,320 tuberculosis patients from March 2010 to March 201 in Iran. Data was gathered from the 345 sections of Iran by Ministry of Health and Medical Education and Statistical Center of Iran. The jittering method was applied for smoothing, and then, the quantile regression for count data was fitted. The AIC was used to compare the fitness of quantile regression for count data model and Poisson log-linear model. The R (3.0.1) software and Quantreg and AER packages were used for all analysis and modeling of the data. Results: The results of fitting the quantile regression for count data showed that in all percentiles, the more increase in immigration rate, illiteracy rate, unemployment and urbanization rates, the more tuberculosis morbidity rate was increased. The maximum increase of tuberculosis due to immigration rate, urbanization rate, unemployment rate, and illiteracy rate was in 95th percentile (β=0.315), 85'Th percentile (β=0.162), 75'Th percentile (β=0.114), and 95'Th percentile (β=0.304), respectively. For 50th percentiles and higher percentiles, with increasing the sum of physicians to the number of population, the tuberculosis morbidity rate was decreased, and the maximum decrease was in 95'Th percentile (β=-0.1). For all percentiles, the AIC showed that quantile regression for count data had been a better fit to data. Conclusion: With respect to the relationship between socioeconomic factors and TB rate, health care observers should pay close attention to improving these factors in Iran to reduce the TB mortality and morbidity