Phenotypic and genotypic evaluation of fluoroquinolone resistance in clinical isolates of Staphylococcus aureus in Tehran

Background: Fluoroquinolones are broad-spectrum antibiotics widely used in the treatment of bacterial infections such as Staphylococcus aureus isolates. Resistance to these antibiotics is increasing. Material/Methods: The occurrence of mutations in the grlA and gyrA loci were evaluated in 69 fluoroquinolone-resistant S. aureus isolates from 2 teaching hospitals of Tehran University of Medical Sciences. Results: Out of the 165 S. aureus isolates, 87 (52.7) were resistant to methicillin and 69 (41.8) were resistant to fluoroquinolone. Fluoroquinolone-resistant S. atoms isolates had a mutation at codon 80 in the grlA gene and different mutational combinations in the gyrA gene. These mutational combinations included 45 isolates at codons 84 and 86,23 isolates at codons 84,86 and 106 and 1 isolate at codons 84, 86 and 90. Fluoroquinolone-resistant S. aureus isolates were clustered into 33 PFGE types. Conclusions: The findings of this study show that the fluoroquinolone-resistant S. aureus strains isolated in the teaching hospitals in Tehran had multiple mutations in the QRDRs region of both grlA and gyrA genes

Regulation of connexin 43 and microRNA expression via β2-adrenoceptor signaling in 1321N1 astrocytoma cells

Connexin 43 (Cx43) is the main gap junction protein in astrocytes and exerts the same effects on growth inhibition in astrocytoma and glioma as microRNA-146a (miR-146a) in glioma. β2-adrenergic receptor (AR) signaling modulates Cx43 expression in myocytes via components downstream of protein kinase A (PKA) and exchange protein directly activated by cAMP (Epac). However, it remains to be elucidated how expression of Cx43 is modulated in astrocytes. In the present study, 1321N1 astrocytoma cells were treated with β2-AR signaling agents in order to evaluate the expression of Cx43 and miRNAs. RNA and protein were extracted from the cells for use in reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. The results revealed that clenbuterol increased miR-146a level and upregulated Cx43 expression via cAMP/PKA at the mRNA and protein level. Pre-inhibition of adenyl cyclase decreased expression of Cx43 and miR-146a. PKA activation and overexpression of miR-146a in A-1321N1 cells increased the expression of Cx43. β2-AR stimulation and 6Bnz, a PKA activator, suppressed oncomiRs miR-155 and miR-27a, while 8-(4-chlorophenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate, an Epac activator, increased their levels. The current findings demonstrated that β2-AR signaling has growth inhibitory effects via modulation of the cAMP/PKA pathway in A-1321N1 cells through increasing the expression level of Cx43 and miR-146a as well as decreasing miR-155 and miR-27a levels. Thus, stimulation of the β2-AR and PKA signaling pathway may be a useful approach for astrocytoma therapy

Combinations of β-lactam or aminoglycoside antibiotics with plectasin are synergistic against methicillin-sensitive and methicillin-resistant Staphylococcus aureus.

Bacterial infections remain the leading killer worldwide which is worsened by the continuous emergence of antibiotic resistance. In particular, methicillin-sensitive (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) are prevalent and the latter can be difficult to treat. The traditional strategy of novel therapeutic drug development inevitably leads to emergence of resistant strains, rendering the new drugs ineffective. Therefore, rejuvenating the therapeutic potentials of existing antibiotics offers an attractive novel strategy. Plectasin, a defensin antimicrobial peptide, potentiates the activities of other antibiotics such as β-lactams, aminoglycosides and glycopeptides against MSSA and MRSA. We performed in vitro and in vivo investigations to test against genetically diverse clinical isolates of MSSA (n = 101) and MRSA (n = 115). Minimum inhibitory concentrations (MIC) were determined by the broth microdilution method. The effects of combining plectasin with β-lactams, aminoglycosides and glycopeptides were examined using the chequerboard method and time kill curves. A murine neutropenic thigh model and a murine peritoneal infection model were used to test the effect of combination in vivo. Determined by factional inhibitory concentration index (FICI), plectasin in combination with aminoglycosides (gentamicin, neomycin or amikacin) displayed synergistic effects in 76-78% of MSSA and MRSA. A similar synergistic response was observed when plectasin was combined with β-lactams (penicillin, amoxicillin or flucloxacillin) in 87-89% of MSSA and MRSA. Interestingly, no such interaction was observed when plectasin was paired with vancomycin. Time kill analysis also demonstrated significant synergistic activities when plectasin was combined with amoxicillin, gentamicin or neomycin. In the murine models, plectasin at doses as low as 8 mg/kg augmented the activities of amoxicillin and gentamicin in successful treatment of MSSA and MRSA infections. We demonstrated that plectasin strongly rejuvenates the therapeutic potencies of existing antibiotics in vitro and in vivo. This is a novel strategy that can have major clinical implications in our fight against bacterial infections

Study of Pseudomonas Aeroginosa resistance to Penicillines, Cephalosporins and Aminoglycosides

Drug therapy and prophylaxy in infectious diseases, from hygienic and economical point of view, are very important. Infections caused by pseudomonas aeroginosa were particularly severe, with high mortality rates. In the recent years pseudomonas aeroginosa continued to cause the most severe, life-thereating infections in burned patients, in spite of the introduction of a wide variety of antibiotics advised specifically for their anti pseudomonal activity. The aim of this study, in which many cases of ps.aeroginosa infections are assessed is to identify the drug resistance of this bacteria to penicillines, cephalosporins and aminoglycosides by antibiotic sensitivity test (disk ager diffusion). Results as percent of resistance to each antibiotic were 89% to carbenicillin, 55% to piperacillin, 89% to mezlocillin, 89.5% to ticarcillin+clavulonic acid, 85% to ceftriaxone, 95% to tobramycin, 5% of all isolates were not sensitive to any antibiotics

Shear bond strength; Blood contamination; Prompt L-Pop, Composite resin; Compomer

Statement of Problem: Root canal cleaning and elimination of the source of infection are the most important basis and the main requirements for successful root treatment since the main cause of failure in root treatment is the permeation of bacteria or their products into the periapical tissues. Nowadays, the newly designed and prcsented instruments for canal instrumentation can improve root treatment. Purpose: The aim of this study was to compare the decrease in the number of intracanal Enterococcus-faecalis (Ef) among three mechanical instrumentation methods: manual (K-type) and rotary (Race, Profile). Materials and Methods: In this experimental study, 30 single rooted teeth were selected. Three cases were considered as negative and three cases as posetive controls and 24 remainder cases were divided into three experimental groups. All root canals were inoculated by Ef and samples were taken from all canals to prepare microbial cultures. The three instrumentation procedures were: - Crown- down technique with K-type manual system file - Crown- down technique with Profile rotary system - Crown- down technique wiht Race rotary system After instrumentation, microbial cultures were taken from root canals and the reduction rate of bacteria were evaluated and compared using one way ANOVA test with P<0.05 as the limit of significance. Results: There was no significant difference among the three instrumentation procedures regarding bacterial elimination. Conclusion: According to the finding of this study, K-type manual file, Profile and Race rotary systems, all can be used in canal instrumentaion. However, since manual files are more accessible and require less equipment compared with rotary systems, and since the ability of all of these methods is identical regarding bacterial elimination, manual files can be used in straight canal instead of rotary systems

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

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

An In-vitro Evaluation of the Ability of 5.25% NaOCl in the Elimination of Enterococcus Faecalis from Root Canal

Statement of Problem: Sodium hypochlorite (NaOCl) have been widely used as an irrigant since it has been introduced to endodontics by walker in 1936, because of its bleaching, deodorizing and tissue dissolving properties. It should be used clinically in concentrations of 3% to 5%.Purpose: The aim of this study was to evaluate the effectiveness of the NaOCl to eliminate Enterococcus faecalis (E.f) from root canals in comparison with Normal saline.Materials and Methods: In an interventional study forty freshly extracted single canal human teeth were chosen. They were sectioned at the CEJ, instrumented and Sterilized.Then they were contaminated with E.f solution and incubated. These samples divided intotwo groups randomly. Root canals were irrigated and filled with 5.25% NaOCl for five minutes in group one, and with normal saline in group two. Then samples were obtained from canals with sterile paper points and cultured for four days. The appearance of turbidityin cultured solutions was the indication to of E.F presence. In order to confirm the specific presence of E.F, three complementary microbiologic tests were applied.Results: All cultures obtained from NaOCl group were negative and all of normal saline group were positive.Conclusion: these results indicate the ability of 5.25% NaOCl to eliminate E.F in prepared root canals with wide diameter

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