Evaluation of cellular immunogenicity of recombinant cytochrome p450 cyp141 protein of Mycobacterium tuberculosis in human and mouse model.

Mycobacterium tuberculosis (Mtb) is still considered one of the unsolved problems for the World Health Organization Identifying and selecting an immunogenic antigen capable of generating specific immune responses is generally the goal of all studies being carried out in to designing new vaccines. Accordingly, the present study was conducted to evaluate the immunogenicity of a M. tuberculosis recombinant protein which exist in the regions of the bacterium genome and may be an immunogenic protein. Immunogenicity of purified proteins was measured by PBMC and mouse spleen lymphocytes culturing methods using ELISA after an appropriate amount of time of incubation with Recombinant cytochrome P450 CYP141 protein. Cellular immune responses were determined and compared by measuring IFN-γ and IL4 in human, and mouse groups. The results revealed a high level of IFN-γ in PPD + individuals and the mice immunized with protein and adjuvant. Recombinant cytochrome P450 CYP141 protein proved capable of generating an immune response in mice and people with a history of previous encounters with Mycobacterium tuberculosis bacteria. It, could be considered a tuberculosis vaccine candidate in order to induce a specific effective immune response in both mice and humans.

Frequency of virulence factors in Helicobacter pylori-infected patients with gastritis.

UNLABELLED: The outcome of Helicobacter pylori infection has been related to specific virulence-associated bacterial genotypes. The vacuolating cytotoxin (vacA), cagA gene, oipA and babA2 gene are important virulence factor involving gastric diseases. The objective of this study was to assess the relationship between virulence factors of H. pylori and histopathological findings. MATERIAL AND METHODS: Gastroduodenoscopy was performed in 436 dyspeptic patients. Antrum biopsy was obtained for detection of H. pylori, virulence factors and for histopathological assessment. The polymerase chain reaction was used to detect virulence factors of H. pylori using specific primers. RESULTS: vacA genotypes in patients infected with H. pylori were associated with cagA, iceA1 and iceA2. In the patients with H. pylori infection there was a significant relationship between cagA positivity and neutrophil activity (P = 0.004) and chronic inflammation (P = 0.013) and with H. pylori density (P = 0.034). Neutrophil infiltration was found to be more severe in the s1 group than in the s2 group (P = 0.042). Also was a significant relationship between oipA positivity and neutrophil activity (P = 0.004) and with H. pylori density (P = 0.018). No significant relationships were observed between other vacA genotypes and histopathological parameters. CONCLUSION: H. pylori strains showing cagA, vacA s1 and oipA positivity are associated with more severe gastritis in some histological features but virulence factors of H. pylori do not appear to determine the overall pattern of gastritis.

The frequency of efflux pump genes expression in Acinetobacter baumannii isolates from pulmonary secretions.

Acinetobacter baumannii is an important opportunistic pathogen, and the cause of nosocomial infections worldwide in recent decades. Efflux pumps are considered as the important causes of multidrug resistance of A. baumannii. The aim of this study was to determine the frequency of efflux pump genes, and evaluate the antibiotic effect of Tigecycline on the expression of adeB gene in isolates of multidrug-resistant. A. baumannii. 70 isolates of A. baumannii were collected and confirmed by biochemical and molecular tests. Antibiotic resistance (Ciprofloxacin, Trimethoprim-sulfamethoxazole, and Tigecycline) was performed based on the minimum inhibitory concentration (MIC) method. Then, the effect of Carbonyl cyanide m-chlorophenyl hydrazone inhibitor (CCCP) on isolates was investigated and the frequency of adeB, adeG, adeJ and abeM genes were examined by PCR for isolates with reduced in MIC titer. Also, the antibiotic effect of Tigecycline on adeB gene expression in A. baumannii isolates was analyzed by Real-Time PCR. The antibiotic resistance for Ciprofloxacin, Trimethoprim-sulfamethoxazole, and Tigecycline was 97.1%, 95.8% and 37.2%, respectively. Following CCCP inhibitor use, the MIC titer had a decrease in MIC titer containing CCCP inhibitor was 64.3% for Ciprofloxacin, 51.5% for Trimethoprim-sulfamethoxazole and 50% for Tigecycline. The frequencies of genes associated with adeB, adeG, adeJ and abeM efflux pump were 100%, 92.8%, 86% and 98.5%, respectively. Real-Time PCR results showed a correlation between the antibiotic effects of Tigecycline on adeB gene expression. The antibiotic resistance of the isolates was relatively high. The isolates were resistant to Ciprofloxacin and Trimethoprim-sulfamethoxazole antibiotics, while more sensitive to Tigecycline. Also, efflux pump genes, which are the antibiotic resistance factors of A. baumannii, are frequently high in the isolates but it seems that isolates use other effluxe pumps than RND family to exit tigecycline.

Isolation and identification of specific bacteriophages against methicillin-resistant Staphylococcus aureus, extended-spectrum beta-lactamases-producing Escherichia coli, extended-spectrum beta-lactamases-producing Klebsiella pneumoniae, and multidrug-resistant Acinetobacter baumanniiin vitro.

Drug resistance of methicillin-resistant Staphylococcus aureus, extended-spectrum beta-lactamases-producing Escherichia coli and Klebsiella pneumoniae and multidrug-resistant Acinetobacter baumannii are also cited as one of the most important causes of community and hospital acquired infections. Phage therapy can be used as a therapeutic method for the treatment of infections caused by these bacteria. The aim of this study was to isolate bacteriophages from municipal wastewater and assess their effects against drug resistant bacterial strains. The single agar layer technique was used to investigate the bacteriolytic effect of bacteriophages. Then, the double agar layer technique was used to observe phage plaques and the transmission electron microscopy was used to study the morphology of the bacteriophages. Transparent plaque formation in a double agar layer test of methicillin-resistant S. aureus and extended-spectrum beta-lactamases-producing E. coli and K. pneumoniae indicated the lysis of bacterial cells by isolated bacteriophages. No bacteriophage against A. baumannii was isolated from municipal wastewater. The morphology of these bacteriophages was also identified by electron microscopy. The results of this study showed that bacteriophages act specifically and due to the increasing level of antibiotic resistance, phage therapy as a new treatment can open a new horizon for the treatment of multidrug resistant bacteria.

Determination of CagA EPIYA motif in Helicobacter pylori strains isolated from patients with digestive disorder.

This study was conducted to identify patterns of cagA EPIYA motifs in H. pylori strains isolated from patients with gastrointestinal diseases in Hospitals of Shahrekord, and investigate the association between these biomarkers and clinical outcomes of gastrointestinal diseases due to H. pylori. In this study, 253 patients with gastrointestinal diseases were studied within 1395-1396. Histopathological investigations and urease test showed that 207 isolates were H. pylori-positive. Then, screening using a molecular technique, PCR, confirmed that 159 isolates had cagA. Finally, the pattern and prevalence of the motifs were determined by PCR and identified a number of motifs were sequenced. Results of this study showed that the pattern of motifs was as follows: ABC (140 isolates) (93/7%), ABCC (6 isolates) (3/77%), ABCCC (4 isolates) (2/5%), AB (7 isolates) (4/4%), AC (1 isolate) (0/6%), and BC (1 isolate) (0/6%). Sequencing results showed the presence of changed EPIYA motif in some isolates. CM motif sequence was also seen in all isolates. In this study, no significant association was seen between the prevalence rate of different patterns and clinical symptoms (p = 0.71). There is a slight association between the presence of ABC motifs and the type of digestive disorder (p = 0.056). Results indicated that ABC was the most frequently seen pattern however, in such that positive cases of ABC motifs were more common in gastritis. All isolates had kinase phosphorylation region, and the observed pattern in this region was a generally western type (ABC).

Phenotypic and genotypic detection of metallo-ß-lactamases in A. baumanii isolates obtained from clinical samples in Shahrekord, southwest Iran.

OBJECTIVE: Acinetobacter baumanii is a pathogenic bacterium that is the cause of many nosocomial infections. This study aimed to determine metallo-ß-lactamases (MBL) produced by the A. baumanii isolates obtained from clinical samples in Shahrekord, southwest Iran. RESULTS: A total of 100 A. baumanii were isolated from 250 clinical samples between June 2013 and June 2014. Then, the isolates were identified by biochemical tests, and MBL screening was conducted by the phenotypic tests modified Hodge, EDTA-disk synergy (EDS), combined disk (CD) and AmpC disc after antibiotic sensitivity test. Using PCR technique the bla genes were detected. Eighty-five (85%) isolates were resistant to meropenem and imipenem. Phenotypic tests showed that out of the 100 isolates, 46, 59, 50, 65 and 65 isolates were positive: AmpC disk, CD, EDS, Modified Hodge and E-test MBL respectively. Gene detection by PCR showed that 23 isolates carried the VIM-1 gene and only three isolates carried the IMP-1 gene. The prevalence of metallo-ß-lactamases isolates containing A. baumanii is increasing. Furthermore, the coexistence of various carbapenemases is dominantly act as a major problem. Continuous monitoring of the infections related to these bacteria should be considered to plan an alternative and new therapeutic strategies.

Prevalence and distribution of adhesins and the expression of fibronectin-binding protein (FnbA and FnbB) among Staphylococcus aureus isolates from Shahrekord Hospitals.

OBJECTIVE: One of the most important causes of nosocomial infections is Staphylococcus aureus. The aim of this study was to determine the frequency of these genes and the rate of expression of these genes during nasal colonization among the personnel of Kashani and Hajar hospitals. RESULTS: In this Analytical-descriptive study, 240 nasal swab specimens were collected from personnel of different departments of Kashani and Hajar hospitals in Shahr-e-kord. Nasal specimens were cultured and 110 Staphylococcus strains were isolated. Based on the results, 110 carriers of Staphylococcus aureus were identified. The frequency of clfA, clfB, fnbA and fnbB genes were 36.3%, 86.3%, 7.2% and 43.6% respectively. It was also observed that the fnbA gene showed no expression, but of 95 clfB-positive samples, 73 isolates (76.8%) were expressed clfB gene. This study showed that the abundance of these genes varies in nasal colonization. It was also observed that clfB gene with a high frequency and high expression rate has an important role in nose colonization. These results not only provide insight into the factors involved in S. aureus colonization but also provide potential therapeutic targets.

A Review Study on Punica granatum L.

Punica granatum L (pomegranate) is a deciduous shrub, native to Iran. Nowadays, besides its use as a fruit, its medicinal properties have attracted the interest of researchers of many countries. Pomegranate fruit has medicinal properties such as anti-inflammatory and antibacterial activities. The pomegranate seed oil has inhibitory effect on skin and breast cancers. The pomegranate seed oil has phytoestrogenic compounds and the fruit is rich in phenolic compounds with strong antioxidant activity. Ellagic acid is one of the main components of pomegranate with phenolic structure and antioxidant activity. This review article presents the recently published findings on different aspects of this plant focusing on its medicinal properties.

The FimH Gene in Uropathogenic Escherichia coli Strains Isolated From Patients With Urinary Tract Infection.

BACKGROUND: Urinary tract infections (UTIs) are one of main health problems caused by many microorganisms, including uropathogenic Escherichia coli (UPEC). UPEC strains are the most frequent pathogens responsible for 85% and 50% of community and hospital acquired UTIs, respectively. UPEC strains have special virulence factors, including type 1 fimbriae, which can result in worsening of UTIs. OBJECTIVES: This study was performed to detect type 1 fimbriae (the FimH gene) among UPEC strains by molecular method. MATERIALS AND METHODS: A total of 140 isolated E. coli strains from patients with UTI were identified using biochemical tests and then evaluated for the FimH gene by polymerase chain reaction (PCR) analysis. RESULTS: The UPEC isolates were identified using biochemical tests and were screened by PCR. The fimH gene was amplified using specific primers and showed a band about 164 bp. The FimH gene was found in 130 isolates (92.8%) of the UPEC strains. Of 130 isolates positive for the FimH gene, 62 (47.7%) and 68 (52.3%) belonged to hospitalized patients and outpatients, respectively. CONCLUSIONS: The results of this study indicated that more than 90% of E. coli isolates harbored the FimH gene. The high binding ability of FimH could result in the increased pathogenicity of E. coli; thus, FimH could be used as a possible diagnostic marker and/or vaccine candidate.

Prevalence assessment of magA gene and antimicrobial susceptibility of Klebsiella pneumoniae isolated from clinical specimens in Shahrekord, Iran.

BACKGROUND AND OBJECTIVES: Klebsiella pneumoniae (K. pneumoniae) is an opportunistic microorganism. This study aimed to investigate the presence of magA gene and antimicrobial susceptibility in K. pneumoniae. MATERIALS AND METHODS: clinical specimens were collected from hospitals of Shahrekord, Iran. Bacterial culture, biochemical diagnostic standard test, determination of antibiotic sensitivity, phenotypic testing hypermucoviscosity (HV) and polymerase chain reaction (PCR) was performed for isolation and characterization of K. pneumoniae. RESULTS: 173 samples were positive for K. pneumoniae. The highest and lowest rates of resistance were related to amoxicillin 79.19% and ciprofloxacin 15.60%, respectively. Also 4 samples were positive for magA gene. CONCLUSION: Based on our results, K. pneumoniae strains were resistant to different antibiotics. Knowing how to identify strains of K. pneumoniae, spreading of its virulence and also antimicrobial resistance genes can be useful in treatment of infection caused by this bacterium.

Comparing the effect of clofibrate and phenobarbital on the newborns with hyperbilirubinemia.

The aim of treating hyperbilirubinemia is preventing the serum bilirubin to reach neurotoxic levels, which is done by phototherapy or blood transfusion. However, pharmacological treatments still remain vague. Therefore the effects of adding either clofibrate or phenobarbital on treatment outcomes was evaluated in icteric non-hemolitic newborns. Ninety neonates were divided in three groups. Two groups were prescribed 100 mg/kg clofibrate or 5 mg/kg phenobarbital orally as single dose on arrival, in addition to phototherapy. The control group only received phototherapy. Serum bilirubin was evaluated at the reception and 12, 24, 48 and 72 hours after beginning of drug therapy. Total bilirubin levels decreased in treated groups compared with the control group in all samples taken (12, 24, 48 and 72 hours). Clofibrate effect in decreasing bilirubin level was more prominent (14 % and 32 % after 12 and 72 h respectively). In addition duration of hospitalization and length of phototherapy decreased in clofibrate and phenobarbital groups compared with control group (1.5, 2 days respectively, vs. 2.6 days). Therefore using clofibrate and phenobarbital in icteric neonates are supportive not only by decreasing the serum bilirubin level, but also by lessening the duration of hospitalization and phototherapy. Thus in addition to cost benefits for the patient these drugs can reduce the risks of transfusion, and clofibrate seems more promising in this regard.