THE ROLE OF HISTONE DEMETHYLASE JMJD3 IN THE INFLAMMATORY RESPONSE: GENERATION AND ANALYSIS OF IN VIVO MODELS

Jmjd3, a JmjC family histone demethylase, is quickly induced by the transcription factor NF- kB in response to microbial stimuli. Jmjd3 erases trimethylated lysine 27 in histone H3 (H3K27me3), a histone mark associated with transcriptional repression and involved in lineage determination, differentiation and tissue homeostasis. However, the specific contribution of Jmjd3 induction and H3K27me3 demethylation to innate immunity and inflammation remains unknown. To define this role, we generated gene-targeted mice lacking histone demethylase using standard protocol to see the effect of this specific Histone demethylase depletion on all tissues and in particular in immune system to understand the biological functions of jmjd3 in inflammatory responses.. Strikingly, transcription of most Jmjd3 target genes was unaffected by its deletion, a few hundred genes including IL12b and Ccl5 showed mild to moderate mRNA changes associated with impaired transcription; however, no gene was completely dependent on Jmjd3 for induction. Importantly, most Jmjd3 target genes were not associated with detectable levels of H3K27me3, and the transcriptional effects of Jmjd3 absence in the window of time analyzed were uncoupled from measurable effects on this histone mark. Overall, these data demonstrate that Jmjd3 participates in fine- tuning the transcriptional output of LPS-activated macrophages in a manner that is largely independent of H3K27me3 demetylation

Prevalence of high-level gentamicin-resistant Enterococcus faecalis and Enterococcus faecium in an Iranian hospital

This study was designed to determine the molecular characteristics and antimicrobial resistance of enterococcal strains isolated from patients admitted to an Iranian Hospital. Enterococcal strains were isolated from the burn patients. All strains were screened for genes encoding resistance to aminoglycoside aac(6')-Ie-aph(2'')-Ia, aph (3'), ant (4'), resistance to vancomycin (vanA, vanB), resistance to tetracycline (tetK, tetL, tetM, tetO), and resistance to erythromycin (ermA, ermB, ermC) by PCR and multiplex PCR-based methods. Genetic diversity was evaluated via Random Amplified Polymorphic DNA (RAPD)-PCR. All enterococcal isolates showed complete sensitivity to vancomycin with MIC � 0.5μg/ml. Resistance to gentamicin, tetracycline, erythromycin, ciprofloxacin or quinopristin-dalfopristin was detected, whilst more than 96.2% of isolates were high-level gentamicinresistant (HLGR) and multiple drug resistant. The most prevalent aminoglycoside resistance gene was aac(6')-Ie-aph(2'')-Ia, that was found in 96.2% (26/27) of the isolates. The most prevalent tetracycline resistance genes were tetM, found in 85.1% (23/27) followed by tetL and tetO found in 7.4% (2/27) of the isolates. The ermA and ermB genes were detected in 33.3% (9/27) and 44.4% (12/27) of the isolates respectively. RAPD-PCR analysis yielded 17 distinct profiles among 27 investigated isolates. One cluster of isolates shared the same RAPD pattern, while 16 isolates had unique RAPD pattern. Our study showed that during the examination time period one RAPD genotype was the common type and was disseminated among patients in the burn unit. Interestingly, most of these strains had an identical or very similar antibiotic and gene resistance pattern

Modification of Salmonella Typhimurium Motility by the Probiotic Yeast Strain Saccharomyces boulardii

BACKGROUND: Motility is an important component of Salmonella enterica serovar Typhimurium (ST) pathogenesis allowing the bacteria to move into appropriate niches, across the mucus layer and invade the intestinal epithelium. In vitro, flagellum-associated motility is closely related to the invasive properties of ST. The probiotic yeast Saccharomyces boulardii BIOCODEX (S.b-B) is widely prescribed for the prophylaxis and treatment of diarrheal diseases caused by bacteria or antibiotics. In case of Salmonella infection, S.b-B has been shown to decrease ST invasion of T84 colon cell line. The present study was designed to investigate the impact of S.b-B on ST motility. METHODOLOGY/PRINCIPAL FINDINGS: Experiments were performed on human colonic T84 cells infected by the Salmonella strain 1344 alone or in the presence of S.b-B. The motility of Salmonella was recorded by time-lapse video microscopy. Next, a manual tracking was performed to analyze bacteria dynamics (MTrackJ plugin, NIH image J software). This revealed that the speed of bacterial movement was modified in the presence of S.b-B. The median curvilinear velocity (CLV) of Salmonella incubated alone with T84 decreased from 43.3 µm/sec to 31.2 µm/sec in the presence of S.b-B. Measurement of track linearity (TL) showed similar trends: S.b-B decreased by 15% the number of bacteria with linear tract (LT) and increased by 22% the number of bacteria with rotator tract (RT). Correlation between ST motility and invasion was further established by studying a non-motile flagella-deficient ST strain. Indeed this strain that moved with a CLV of 0.5 µm/sec, presented a majority of RT and a significant decrease in invasion properties. Importantly, we show that S.b-B modified the motility of the pathogenic strain SL1344 and significantly decreased invasion of T84 cells by this strain. CONCLUSIONS: This study reveals that S.b-B modifies Salmonella's motility and trajectory which may account for the modification of Salmonella's invasion

A Large Fraction of Extragenic RNA Pol II Transcription Sites Overlap Enhancers

A substantial fraction of extragenic Pol II transcription sites coincides with transcriptional enhancers, which may be relevant for functional annotation of mammalian genomes

Comparison of virulence factors and biofilm formation among Staphylococcus aureus strains isolated from human and bovine infections

The aim of this study was to find different prevalence of genes involved in the biofilm formation process and to assess the phenotypic and genotypic markers of biofilm formation among Staphylococcus aureus strains isolated from human and bovine infections. In this study, 215 S. aureus strains were collected from human and dairy cow's infections. The biofilm forming capacity of the strains was evaluated using a colorimetric microtiter plate assay. The genes encoding microbial surface components, recognizing adhesive matrix molecules (MSCRAMMs) (ebpS, eno, fib, fnbA, fnbB, cna and bap), and the intracellular adhesion (ica) genes (icaA, and icaD) were targeted by polymerase chain reaction (PCR)-based method. Approximately 70 of the isolates produced biofilm. Among these, 59.3 were producers of weakly adherent biofilms while 34.8 and 5.8 produced moderate and strong biofilms, respectively. The most prevalent gene was icaD found in 88.4 of the isolates, followed by icaA, fib and eno found in 87.9, 75.8 and 75.3 of the isolates, respectively. The bap gene was not detected in any of the isolates. The prevalence of ebpS and fnbA genes among bovine isolates were significantly higher than those in human isolates, whilst the prevalence of cm gene was significantly higher in the human isolates. In this study, a high prevalence of biofilm production was found among S. aureus strains isolated from human and bovine infections. Most biofilm producing isolates were positive for MSCRAMM, icaA, and icaD genes. (C) 2015 Elsevier Ltd. All rights reserved

Multiple-locus variable number of tandem repeats (VNTR) fingerprinting (MLVF) and antibacterial resistance profiles of extended spectrum beta lactamase (ESBL) producing Pseudomonas aeruginosa among burnt patients in Tehran

Extended spectrum p-lactamase (ESBL)-producing trait was present in 48 out of the 112 (42.8) Pseudomonas aeruginosa isolates collected from burn wound infections during a 12-month period. The presence of oxa-10, per-1, veb-1 and ges genes and the multiple-locus variable number of tandem repeats (VNTR) fingerprinting (MLVF) of 112 P. aeruginosa strains were determined by PCR and multiplex PCR. Disk diffusion methods were used to determine the susceptibility of the isolates to antimicrobial agents as instructed by CLSI. All ESBL isolates were resistant to aztreonam, cefepime, cefotaxime, cefpodoxime, ceftazidime, ceftriaxone and ofloxacin. Fewer than 60 of ESBL isolates were resistant to imipenem, meropenem, and piperacillin-tazobactam but more than 90 were resistant to amikacin, ciprofloxacin, levofloxacin, ticarcillin and tobramycin. The most prevalent ESBL genes included oxa-10 (70) and per-1 (50) followed by veb-1 (31.3). The gene encodes GES enzyme did not detect in any isolates. A total of 100 P. aeruginosa strains were typed by MLVF typing method. MLVF produced 42 different DNA banding patterns. These data indicate that different MLVF types infect burn wounds in patients at a hospital in Tehran and also suggest an alarming rate of ESBL-producing isolates in this test location. (C) 2011 Elsevier Ltd and ISBI. All rights reserved

A high prevalence of mupirocin and macrolide resistance determinant among Staphylococcus aureus strains isolated from burnt patients

Infections due to Staphylococcus aureus have become increasingly common among burn patients. The antibiotic resistance profile of S. aureus isolates and inducible resistance against clindamycin were investigated in this study. The presence of mecA gene, mupA gene and macrolide resistance genes were detected using PCR and multiplex-PCR. The resistance rate to methicillin, erythromycin and mupirocin were 58.5, 58 and 40, respectively. The prevalence of constitutive and inducible resistance among macrolide resistant isolates was 75 and 25, respectively. Ninety five percent of the isolates were positive for one or more erm genes. The most common genes were ermA (75), ermC (72) and ermB (69), respectively. The ermA gene predominated in the strains with the inducible phenotype, while ermC was more common in the isolates with the constitutive phenotype. The msrA gene was only found in one MRSA isolate with the constitutive phenotype. A total of 27 isolates (25) carried the mupA gene. All the mupirocin resistant isolates and almost all the erythromycin resistant isolates were also resistant against methicillin which may indicate an outbreak of MRSA isolates with high-level mupirocin and erythromycin resistance in the burn unit assessed. (C) 2011 Elsevier Ltd and ISBI. All rights reserved

Evaluation of biofilm production and characterization of genes encoding type III secretion system among Pseudomonas aeruginosa isolated from burn patients

Pseudomonas aeruginosa is one of the common pathogenic causes of serious infections in burn patients throughout the world. Type III secretion toxins are thought to promote the dissemination of P. aeruginosa from the site of infection, the bacterial evasion of the host immune response and inhibition of DNA synthesis leading to host cell death. A total of 96 isolates of P. aeruginosa were collected from wound infections of burn patients, from April to July 2010. Antimicrobial susceptibility of the isolates were determined by disk agar diffusion method. Polymerase chain reaction (PCR)-based method was used for targeting the genes encoding the type III secretion toxins. The quantitative determination of biofilm-forming capacity was determined by a colorimetric microtiter plate assay. All the isolates were resistant to cefixime and ceftriaxone. More than 90 of the isolates were resistant to amikacin, carbenicillin, cefepime, cefotaxime, cefpodoxime, gatifloxacin, gentamicin, piperacillin/tazobactam, ticarcillin and tobramycin. All the isolates carried the exoT gene, 95 carried exoY, 64.5 carried exoU and 29 carried the exoS gene. Most of the isolates (58) carried both exoY and exoU genes while 24 showed the concomitant presence of exoS and exoY and 1 carried both exoS and exoU. Coexistence of exoS, exoY and exoU was seen in 4 of the isolates. Biofilm formation was seen in more than 96 of the isolates among which 47 were strong biofilm producers, 26 were moderate and 22.9 were weak biofilm formers. In conclusion, the findings of this study show that the genes, particularly the exoU gene, encoding the type III secretion toxins, are commonly disseminated among the P. aeruginosa strains isolated from burn patients. (c) 2012 Elsevier Ltd and ISBI. All rights reserved

CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS STRAINS ISOLATED FROM RAW MILK OF BOVINE SUBCLINICAL MASTITIS IN TEHRAN AND MASHHAD

Staphylococcus aureus is considered one of the most important food borne pathogens. A total of 111 isolates of S. aureus were cultured from raw milk samples during January 2009 to June 2009 from Tehran and Mashhad. The coagulase gene polymorphism and the prevalence of classical enterotoxin genes of S. aureus strains were determined by PCR-RFLP (restriction fragment length polymorphism) and Multiplex-PCR. Disk diffusion method was used to determine the susceptibility of isolates to antimicrobial agents as instructed by Clinical and Laboratory Standards Institute. Sixty-seven of the isolates harboured one or more enterotoxin genes. The most prevalent gene was sec, found in 59 of the isolates. Approximately 8 of the isolates were positive for sea, seb and sed genes. Only one isolate had see gene. The rate of coexistence of enterotoxin genes was 14. All S. aureus isolates were susceptible to ciprofloxacin, gentamicin, imipenem, minocycline, oxacillin and vancomycin. They were resistant to ampicillin (64), penicillin (56), clindamycin (22), tetracycline (22), doxycycline (19), teicoplanin (13), rifampin (2) and tri-methoprim-sulfamethoxazole (2). On the basis of coagulase gene analysis of 111 S. aureus isolates, the PCR products of 56 isolates were digested with Alu I that produced three distinct patterns. These data indicate the high prevalence of enterotoxigenic S. aureus in raw bovine milk in Tehran and Mashhad, and highlight the importance of proper quality control of dairy products for public health