Quinolone resistance and biofilm formation capability of uropathogenic Escherichia coli isolates from an Iranian inpatients' population.

BACKGROUND: Uropathogenic Escherichia coli (UPEC) is a major pathogen of the urinary tract infection (UTI), and biofilm formation is crucial as it facilitates the colonization in the urinary tract. We aimed to investigate the antibiotic susceptibility pattern, biofilm formation capability, distribution of quinolone resistance genes, and phylogenetic groups among UPEC isolates from an Iranian inpatients' community. METHODS AND RESULTS: A collection of 126 UPEC obtained from hospitalized patients with symptomatic UTI at 3 teaching hospitals during 2016 were included. Antibiogram of all isolates against quinolone and fluoroquinolones was performed using the disk diffusion method. Phylogenetic groups and qnr A, B, and S genes were assessed by PCR. Susceptibility pattern showed that more than 50% and 81% of the isolates were resistant to fluoroquinolones and quinolones, correspondingly. The frequency of qnrS and qnrB genes was 22% and 13.5%, correspondingly. Our result indicated no significant association between the presence of fluoroquinolone genes and antibiotic resistance to them. The frequent common phylogroup was B2 (84.1%), followed by D (10.3%), A (3.2%) and B1 (2.4%) groups. Indeed, 80.2% of the isolates were biofilm producers, so that 42.1%, 16.7% and 21.4% of them were classified as weak, moderate and strong producers, respectively. CONCLUSIONS: Our results showed considerable fluoroquinolone and quinolone resistance among UPEC along with a remarkable rate of biofilm-producing isolates from symptomatic hospitalized patients, making them a serious health concern in the region. This survey highlights the need for awareness on quinolone resistance and careful prescription of them by physicians.

High frequency of carbapenem-resistant Enterobacteriaceae fecal carriage among ICU hospitalized patients from Southern Iran.

Objectives: The worldwide emergence of carbapenem-resistant Enterobacteriaceae (CRE) has become a major therapeutic concern to medical institutions. To date, no study has determined the frequency and risk factors of inpatients with CRE fecal carriage in Southern Iran. We studied the features of carbapenemase-producing Enterobacteriaceae (CPE) collected from the central ICU of a university hospital. Materials and Methods: Totally, 173 samples, including 124 stool samples from 46 ICU inpatients on admission and different follow-ups, 9 ICU staff, and 40 environmental samples were included. CRE was identified using microbiological methods. Antimicrobial susceptibility was investigated by using the disk diffusion method and E-test. Carbapenemase producers were detected using the mCIM method. Seven carbapenemase genes were characterized. The genetic relationship among 20 CPE was elucidated by PFGE. Results: The overall fecal carriage rate was 28.2%, while CRE acquisition was 6.1%. CRE were classified as Klebsiella pneumoniae (71.4%), Escherichia coli (23.8%), and Enterobacter aerogenes (4.8%). From 21 CRE, 20 (95.2%) produced carbapenemases, of which 10, 15, 10, 25, 5, and 65% were blaKPC, blaSME, blaIMP, blaVIM, blaNDM and blaOXA-48-positive, respectively. Out of 20 CPE, 14 different PFGE patterns were observed, categorized into six clusters, suggestive of non-clonal spread. No difference between the examined risk factors with CRE carriage was shown. Conclusion: The data indicate a high CRE fecal carriage rate among inpatients. Our findings implicate the widespread of OXA-48 carbapenemase together with heterogeneity among CRE with great concern for dissemination and therapeutic threat. Early diagnosis and monitoring of CRE among inpatients are urgent.

Mesoporous silica nano-adjuvant triggers pro-inflammatory responses in Caco-2/peripheral blood mononuclear cell (PBMC) co-cultures.

The aim of this study was to evaluate the cytotoxicity and immune-stimulatory effect of Mesoporous silica nanoparticle (MSN) Nano-adjuvant on pro-inflammatory cytokines and pattern recognition receptors (PRR) genes expression in Caco-2/PBMC co-culture model. MSNs were synthesized and characterized by scanning electron microscope (SEM), Brunauer Emmett Teller (BET) and Barrett Joyner Halenda (BJH) techniques. The BET specific surface area of MSNs was around 947 m2/g and the total pore volume and average pore diameter were 1.5 cm3/g and 8.01 nm, respectively. At the concentration of 10 µg/mL, MSN showed a low and time-dependent cytotoxicity on Caco-2 cells, while no cytotoxic effect was observed for 0.1 and 1 µg/mL concentrations after 24, 48 and 72 h. The expression of pro-inflammatory cytokines genes (IL-1, IL-8 and TNF-α) in co-cultures treated with different concentrations of MSN showed a dose-dependent significant increase up to 17.44, 2.722 and 4.34 folds, respectively, while the expression augmentation of IL-1 gene was significantly higher than the others. This indicates slight stimulation of intestinal inflammation. Different concentrations of MSN significantly increased TLR4 and NOD2 expression to 4.14 and 2.14 folds, respectively. NOD1 was not affected significantly. It can be concluded that MSN might increase protective immune responses against antigens as a vaccine adjuvant candidate. It seems that stimulation of TNF-α, IL-1, and IL-8 expression in enterocytes probably transpires through the agonistic activity of MSN for TLRs including TLR4, while NOD2-associated signaling pathways are also involved. This study provides an overall picture of MSN as a novel and potent oral adjuvant for mucosal immunity.

Vibrio cholerae toxin coregulated pilus provokes inflammatory responses in Coculture model of Caco-2 and peripheral blood mononuclear cells (PBMC) leading to increased colonization.

The aim of this study was to assess the modulatory effect of TcpA in the expression of CEACAM1 adhesin molecule and IL-1, IL-8, and TNF-α pro-inflammatory cytokines in the Coculture model of Caco-2/PBMC (peripheral blood mononuclear cell) that can mimic the intestinal milieu. The TcpA gene from Vibrio cholerae ATCC14035 was cloned in pET-28a and transformed into Escherichia coli Bl-21. The recombinant TcpA-His6 protein was expressed and purified using Ni-column chromatography. The sequencing of transformed plasmid and Western blot analysis of purified protein confirmed the identity of rTcp. The cytotoxicity of different concentrations of recombinant protein for human colon carcinoma cell line (human colorectal adenocarcinoma cell [Caco-2 cell]) was assessed by MTT assay and showed viability of 92%, 82%, and 70%, for 10 µg/mL of TcpA after 24, 48, and 72 h, respectively. Co-cultures of Caco-2 and PBMCs were used to mimic the intestinal milieu and treated with different concentrations of rTcpA (1, 5, 10, and 50 µg/mL). Our data showed about 2.04-, 3.37-, 3.68-, and 42.7-fold increase in CEACAM1 gene expression, respectively, compared with the nontreated Caco-2/PBMC Coculture. Moreover, the expression of IL-1, IL-8, and TNF-α genes was significantly increased up to 15.75-, 7.04-, and 80.95-folds, respectively. In conclusion, V. cholerae TcpA induces statistically significant dose-dependent stimulatory effect on TNF-α, IL-,1, and IL-8 pro-inflammatory cytokines expression. Of these, TNF-α was much more affected which, consequently, elevated the CEACAM1 expression level in IECs. This suggests that TcpA protein is a critical effector as an inducer of increased adhesion potential of V. cholera as well as inflammatory responses of host intestinal tissue.

Evaluation of flexural strength and antibacterial effect of orthodontic acrylic resins containing Galla chinensis extract

ABSTRACT Objective: To evaluate different concentrations of Galla chinensis extract (GCE) added to poly(methyl methacrylate) (PMMA), which is widely used for fabrication of removable orthodontic appliances, regarding the effectiveness of this herbal extract on antimicrobial effect and flexural strength of PMMA. Methods: Acrylic resin samples containing 0.4%, 0.8% and 1.6% GCE were prepared. Flexural strength was investigated via three-point flexural strength test for the 15 acrylic resin blocks of each concentration. Disk diffusion test was used to evaluate antibacterial effects of incorporating the same concentrations of GCE into acrylic resin. All these three groups were compared with the control group, with no added GCE, regarding flexural strength and antibacterial properties. Results: Comparison of flexural strength between the three study groups and the control group showed significant differences between the groups (P=0.018). However, there was no significant difference between the groups containing GCE. There were significant differences in antimicrobial activity between the four groups (P=0.026). Conclusion: Within the limitations of this study, it is suggested that incorporation of GCE into PMMA would be beneficial for antimicrobial activity and flexural strength of PMMA, but further studies on other physical properties and antimicrobial effects on other bacterial strain would be beneficial prior to clinical investigations.

RESUMO Objetivo: Avaliar se diferentes concentrações de extrato de Galla chinensis (EGC) adicionado ao polimetilmetacrilato (PMMA), que é amplamente utilizado para a fabricação de aparelhos ortodônticos removíveis, interferem no efeito antimicrobiano desse extrato e na resistência à flexão do PMMA. Métodos: Foram preparadas amostras de resina acrílica com concentrações de 0,4%, 0,8% e 1,6% de EGC. Para a avaliação da resistência à flexão, utilizou-se o teste de flexão em três pontos para as 15 amostras de resina em cada concentração. O teste de disco-difusão foi utilizado para avaliar os efeitos antibacterianos da incorporação das mesmas concentrações de EGC na resina acrílica. Esses três grupos foram comparados ao grupo controle, sem adição do EGC, em relação à resistência à flexão e quanto às propriedades antimicrobianas. Resultados: As comparações dos três grupos com o grupo controle mostraram diferenças significativas (p=0,018) para a resistência à flexão. Entretanto, não houve diferença significativa entre os grupos contendo EGC. Foram encontradas diferenças significativas na atividade antimicrobiana entre os quatro grupos (p=0,026). Conclusão: Dentro das limitações desse estudo, parece que a incorporação de EGC no PMMA seria benéfica para a atividade antimicrobiana e a resistência à flexão do PMMA. Porém, estudos adicionais sobre outras propriedades físicas e sobre os efeitos antimicrobianos contra diferentes cepas de bactérias seriam interessantes antes de se fazer pesquisas clínicas.

Modulatory effect of Vibrio cholerae toxin co-regulated pilus on mucins, toll-like receptors and NOD genes expression in co-culture model of Caco-2 and peripheral blood mononuclear cells (PBMC).

Vibrio cholerae, the causative agent of cholera, tend to colonize the small intestine as a Gram-negative pathogen. The intestinal mucus layer forms mucin physical barrier, consisted of high molecular weight proteins. Regarding the role of toxin-coregulated pilus (TCP) as one of the most important colonization factors of V. cholerae, this experimental study was designed to determine the role of TcpA in induction of mucin production and its regulatory effect on innate immunity molecules including toll like receptors (TLRs) and Nucleotide-binding oligomerization domain-containing proteins (NODs) using Caco2- PBMC co-cultures as an interactive model. The rTcpA protein of V. cholerae was expressed in BL21 Escherichia coli, purified using Ni-column chromatography and confirmed by western blotting. Nontoxic doses of rTcpA was determined on Caco-2 cell lines and different concentrations of rTcpA (1, 5, 10 and 50 µg/mL) showed a statistically significant effect on the expression of muc genes (MUC3 and MUC4) in a dose-dependent manner. This finding is supposed to facilitate physical adhesion and colonization of V. cholerae in intestinal lumen. The rTcpA moderately stimulated the expression of tlr4 and overexpressed tlr1, both of which are supposed to induce a mucosal protective response against bacterial infection. NOD2 was significantly increased which suggests that it may contribute in pro-inflammatory responses observed in cholera disease. No change in NOD1 expression was seen which might be attributed to the non-invasive nature of V. cholerae as an intestinal pathogen. In conclusion, the rTcpA protein of V. cholerae showed a statistically significant modulatory effect on the human gut epithelium gene expression which would help promising protection in prophylaxis applications.

The first report of Enterobacter gergoviae carrying bla NDM-1 in Iran.

OBJECTIVES: Prompt detection of extended-spectrum ß-lactamases (ESBL) and carbapenemase-producing enterobacteriaceae is crucial for infection prevention and control strategies. The present study aimed to characterize the ESBL and carbapenemase genes among Enterobacter isolates from an Iranian inpatient population. MATERIALS AND METHODS: A total of 96 Enterobacter isolates obtained from inpatients between June 2016 and March 2017, were identified by the conventional microbiological methods and diagnostic kits. Antimicrobial susceptibility pattern was performed using the disk diffusion method. The ESBL and carbapenemase genes were screened using polymerase chain reaction (PCR). RESULTS: All clinical isolates of Enterobacter were classified as E. gergoviae (52, 54.2%), E. aerogenes (34, 35.4%), E. cloacae (7, 7.3%), Cronobacter (E). sakazakii (3, 3.1%). The highest and lowest antimicrobial resistance rates were observed against ampicillin (93.8%) and imipenem (21.9%). High prevalence of multi-drug resistance (MDR=96.9%) was substantial. Of the 96 Enterobacter isolates, 35 (36.5%) and 28 (29.2%) were phenotypically ESBL-positive and non-susceptible carbapenem, respectively. Overall, the frequency of evaluated genes was as follows: blaCTX-M =25 (26%), blaTEM =30 (31.3%), blaSHV =12 (12.5%), blaIMP =3 (3.1%), blaVIM =0 (0%), blaNDM =8 (8.3%), and blaKPC =0 (0%). CONCLUSION: In this study, we report for the first time the presence of E. gergoviae harboring blaNDM from an Iranian population. Regarding the increase of MDR Enterobacter spp. in our region, strict hygiene rules will be needed to control the quick spread of ESBL and carbapenemase-producing Enterobacter isolates in healthcare facilities of developing countries.

Evaluation of flexural strength and antibacterial effect of orthodontic acrylic resins containing Galla chinensis extract.

OBJECTIVE: To evaluate different concentrations of Galla chinensis extract (GCE) added to poly(methyl methacrylate) (PMMA), which is widely used for fabrication of removable orthodontic appliances, regarding the effectiveness of this herbal extract on antimicrobial effect and flexural strength of PMMA. METHODS: Acrylic resin samples containing 0.4%, 0.8% and 1.6% GCE were prepared. Flexural strength was investigated via three-point flexural strength test for the 15 acrylic resin blocks of each concentration. Disk diffusion test was used to evaluate antibacterial effects of incorporating the same concentrations of GCE into acrylic resin. All these three groups were compared with the control group, with no added GCE, regarding flexural strength and antibacterial properties. RESULTS: Comparison of flexural strength between the three study groups and the control group showed significant differences between the groups (P=0.018). However, there was no significant difference between the groups containing GCE. There were significant differences in antimicrobial activity between the four groups (P=0.026). CONCLUSION: Within the limitations of this study, it is suggested that incorporation of GCE into PMMA would be beneficial for antimicrobial activity and flexural strength of PMMA, but further studies on other physical properties and antimicrobial effects on other bacterial strain would be beneficial prior to clinical investigations.

Viral etiology of acute respiratory infections in children in Southern Iran.

INTRODUCTION: Prevalence of influenza A virus (Flu-A), respiratory syncytial virus (RSV), and human metapneumovirus (hMPV) was assessed in children with acute respiratory infections (ARIs). METHODS: Nasopharyngeal aspirates and throat swabs were subjected to real-time polymerase chain reaction (PCR) to detect RSV and Flu-A and to conventional PCR to detect hMPV. RESULTS: Of the 156 children assessed, 93 (59.6%) carried at least one virus, with 35.9% positive for RSV, 14.1% for hMPV, and 9.6% for Flu-A. The prevalence of co-infections was 2.6%. CONCLUSIONS: The high detection rate may reflect increased sensitivity of real-time PCR compared to traditional PCR and viral culture.

The Occurrence of Nosocomial Pathogens on Cell Phones of Healthcare Workers in an Iranian Tertiary Care Hospital.

BACKGROUND: Cell phones have become one of the necessary means of life and they are commonly used almost everywhere by every population. Colonized microorganisms on cell phones can be easily cross-transmitted. Given the widespread prevalence of nosocomial infections, this study aimed to determine the frequency of bacterial contamination and antibiotic resistance in cell phones of healthcare workers (HCWs) in a tertiary care hospital, from southwest of Iran. In this cross-sectional study conducted between April and June 2016, sampling were performed from cell phones of 25 nurses and 75 medical students. METHODS: Samples were collected from each cell phone by a moistened cotton swap dipped in normal saline prior and after decontamination with available alcohol-based handrubs. Identification of bacterial isolates was performed by conventional microbiologic methods. Antibiotic susceptibility pattern of the isolates was determined using the disk diffusion method. The contamination rates of cell phones prior and after disinfection were 88% and 52%, respectively. Ninety-nine (71.2%) out of 139 isolated distinct bacterial colonies prior to cleaning were potentially nosocomial pathogens. Of them, staphylococci (88.9%) were the most prevalent bacteria, in which 40.9% were methicillin-resistant isolates. The majority of Gram-positive and - negative isolates were susceptible to the tested antimicrobials. Totally, contamination rate of cell phones was significantly reduced after decontamination. Regular disinfection of the hands and cell phones was significantly associated with reduction of colonization of the methicillin-resistant isolates. RESULT & CONCLUSION: These findings emphasize the restricted use of cell phones and encourage the higher compliance with hygienic practices in hospitals to reduce the risk of nosocomial infections.

Viral etiology of acute respiratory infections in children in Southern Iran

Abstract INTRODUCTION: Prevalence of influenza A virus (Flu-A), respiratory syncytial virus (RSV), and human metapneumovirus (hMPV) was assessed in children with acute respiratory infections (ARIs). METHODS: Nasopharyngeal aspirates and throat swabs were subjected to real-time polymerase chain reaction (PCR) to detect RSV and Flu-A and to conventional PCR to detect hMPV. RESULTS: Of the 156 children assessed, 93 (59.6%) carried at least one virus, with 35.9% positive for RSV, 14.1% for hMPV, and 9.6% for Flu-A. The prevalence of co-infections was 2.6%. CONCLUSIONS: The high detection rate may reflect increased sensitivity of real-time PCR compared to traditional PCR and viral culture.

Distribution of virulence genes and their association with antimicrobial resistance among uropathogenic Escherichia coli isolates from Iranian patients.

BACKGROUND: Urinary tract infections (UTIs) are one of the most frequent diseases encountered by humans worldwide. The presence of multidrug-resistant (MDR) uropathogenic Escherichia coli (UPEC) harboring several virulence factors, is a major risk factor for inpatients. We sought to investigate the rate of antibiotic resistance and virulence-associated genes among the UPECs isolated from an Iranian symptomatic population. METHODS: A total of 126 isolates from inpatients with UTI from different wards were identified as UPEC using the conventional microbiological tests. After identification of UPECs, all the isolates were subjected to antimicrobial susceptibility test and polymerase chain reaction (PCR) to identify the presence of 9 putative virulence genes and their association with the clinical outcomes or antimicrobial resistance. RESULTS: The data showed that the highest and the lowest resistance rates were observed against ampicillin (88.9%), and imipenem (0.8%), respectively. However, the frequency of resistance to ciprofloxacin was found to be 55.6%. High prevalence of MDR (77.8%) and extended-spectrum ß-lactamase (ESBL) (54.8%) were substantial. PCR results revealed the frequency of virulence genes ranged from 0 to 99.2%. Among 9 evaluated genes, the frequency of 4 genes (fimH, sfa, iutA, and PAI marker) was > 50% among all the screened isolates. The iutA, pap GII, and hlyA genes were more detected in the urosepsis isolates with significantly different frequencies. The different combinations of virulence genes were characterized as urovirulence patterns. The isolates recovered from pyelonephritis, cystitis, and urosepsis cases revealed 27, 22, and 6 virulence patterns, respectively. A significant difference was determined between ESBL production with pap GII, iutA, and PAI marker genes. CONCLUSIONS: Our study highlighted the MDR UPEC with high heterogeneity of urovirulence genes. Considering the high rate of ciprofloxacin resistance, alternative drugs and monitoring of the susceptibility profile for UPECs are recommended.

Phenotypic and genotypic characterization of macrolide, lincosamide and streptogramin B resistance among clinical isolates of staphylococci in southwest of Iran.

OBJECTIVE: The present study aimed to determine the phenotypic and genotypic profile of macrolide, lincosamide and streptogramin B (MLSB) resistance in clinical isolates of staphylococci. RESULTS: This cross-sectional study was conducted on 164 non-duplicated staphylococci isolates collected during August 2015 to February 2016 from two tertiary care hospitals in Shiraz, southwest of Iran. Of the 164 isolates, 86 erythromycin-resistant isolates consist of 35 Staphylococcus aureus and 51 coagulase negative staphylococci (CoNS) were included in the study. Of the 35 S. aureus, the prevalence of cMLS (constitutive), iMLS (inducible), and MS phenotypes were found 82.9%, 8.6% and 8.6%, respectively. Among 51 CoNS, the frequencies of cMLS, iMLS, and MS phenotypes were detected 66.7%, 11.8% and 21.6%, respectively. Among S. aureus isolates, the predominant genes were ermC in 82.9% isolates, followed by ermA in 57.1% and msrA in 28.6% of isolates. Among CoNS isolates, the most frequent genes were diagnosed ermC in 70.6% isolates followed by msrA in 68.6% and ermA in 11.8% of isolates. In conclusion, regarding the presence of MLSB resistance in our region, diagnosis of this resistance type on a routine basis in staphylococcal clinical isolates is of particular importance.

A rare case of complicated pericardial effusion with Elizabethkingia meningoseptica from Iran.

Infections due to Elizabethkingia meningoseptica, a Gram-negative oxidative bacterium are frequently founded in neonatal and immunocompromised individuals. The notable characteristic of this organism is its multi-drug resistance to common antibiotics used for infections caused by Gram-negative bacteria. We report a rare case of complicated pericardial effusion due to E. meningoseptica in a 2-year-old boy, who was admitted with chief complaints of fever and tachypnea (mentioned by his parents) and suffered from a rare lung malignancy (lymphangioleiomyomatosis). He was successfully treated with vancomycin.  E. meningoseptica infection is a rare situation in immunocompetent hosts, and we concluded that this infection was probably originated from device medicine or even hands of healthcare workers.

Low Rate of babA2 Genotype among Iranian Helicobacter pylori Clinical Isolates.

INTRODUCTION: The Blood Group Antigen-Binding Adhesion (babA), Outer Inflammatory Protein (oipA) and Sialic Acid-Binding Adhesin (sabA) as outer membrane proteins involved in Helicobacter pylori adherence to gastric mucosa have been suggested to have a role in the pathogenesis. AIM: To investigate the frequency of H. pylori isolates babA2, oipA and sabA genes in Iranian dyspeptic patients. MATERIALS AND METHODS: DNAs were extracted from H. pylori -positive cultures taken from 100 different dyspeptic patients. Genotyping was performed by Polymerase Chain Reaction (PCR), using the specific primers for babA2, oipA and sabA genes. Chi square test was used to investigate association between variables, p<0.05 was considered statistically significant. RESULTS: All (100%) isolates possessed oipA and sabA genotypes, whereas babA2 was detected in 22% of isolates. There was no significant relationship between presence of genes with clinical outcome. The combined genotype oipA +/sabA +/ babA2- was correlated with gastritis. The rate of babA2 genotype in our isolates was lower than other Iranian reports. CONCLUSION: Frequency of babA2 genotype among H. pylori isolates from Southwest of Iran is considerably less than other regions of Iran. Due to heterogeneity of H. pylori strains in different geographic regions, further work will be needed to understand the role of these virulence genes in H. pylori pathogenesis and their possible association with disease outcome.

High frequency of hopQ genotypes among Iranian Helicobacter pylori clinical isolates.

Different virulence factors are involved in Helicobacter pylori pathogenesis. H. pylori outer membrane proteins are a family of virulence factors that have diverse members. HopQ (H. pylori outer membrane protein) is the largest of them that contains types I and II alleles. The role of hopQ is not exactly known, but it has been considered in H. pylori adhesion and colonization. The aim of this study was to determine the frequency of hopQ genotypes among H. pylori isolates obtained from patients with gastroduodenal disorders and their association with the clinical outcome. The DNA of 100 H. pylori clinical isolates was investigated by polymerase chain reaction (PCR) method using specific primers for determining the hopQI and hopQII genotypes. hopQI was present in 35%, while hopQII was positive in 55% of the isolates. Amongst the gastritis subjects, the rate of hopQII compared to hopQI was higher, and a statistically significant difference was found between hopQII genotype and the clinical outcome. With respect to the significant difference between the hopQ genotype and clinical outcome in our clinical isolates, it seems that this genotype is a useful marker for evaluating its association with H. pylori-related diseases.

High rate of A2142G point mutation associated with clarithromycin resistance among Iranian Helicobacter pylori clinical isolates.

This study aimed to investigate the clarithromycin resistance and its associated molecular mechanisms among Helicobacter pylori isolates from dyspeptic patients in Shiraz, Iran. From January to May 2014, 100 H. pylori strains were isolated from patients with gastroduodenal disorders. The resistance to clarithromycin was quantitatively evaluated, using Epsilometer (E-test) method. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed on all the isolates to detect A2143G and A2142G mutations in 23S rRNA gene. The H. pylori isolation rate was found to be 31.4%. E-test showed that 20% of isolates were resistant to clarithromycin (MIC ≥ 1 mg/L). MIC of clarithromycin ranged between 0.016 and 24 mg/L. Findings of PCR-RFLP showed that the A2142G was the most (90%) frequently point mutation, followed by the A2143G (10%). No statistically significant difference was found between H. pylori clarithromycin resistance point mutations and patients' gender or age. To the best of our knowledge, this is the first report of high frequency of A2142G point mutation in Iran and probably in other regions of the world. Considering the increasing trend of H. pylori resistance to clarithromycin due to these mutations, it is crucial to investigate the new therapeutic approaches against H. pylori infection.

The Frequency of Exotoxin A and Exoenzymes S and U Genes Among Clinical Isolates of Pseudomonas aeruginosa in Shiraz, Iran.

Pseudomonas aeruginosa as an opportunistic pathogen produces several virulence factors. The most important of these factors are exotoxin A and type III secretion system (T3SS). The aim of this study was to determine the frequency of toxA, exoU and exoS genes among clinical isolates of P. aeruginosa. In this cross-sectional study from September 2011 to February 2012, 156 P. aeruginosa isolates were recovered from different clinical samples. Susceptibility testing against 10 antibiotics was performed on individual isolates by the disc diffusion method according to CLSI guidelines. Extracted DNA was subjected to PCR assay for determining the presence of toxA, exoU and exoS genes. Overall, the frequency of toxA, exoU and exoS genes were 90.4%, 66.7% and 65.4%, respectively. All of the abdominal and eye isolates were exoS (+). The frequency of exoS (+)/exoU (-) and exoS (-)/exoU (+) genotypes was estimated 19.2% and 16.2%, respectively. Indeed, genotypes exoS (+)/exoU (+) and exoS (-)/exoU (-) were found with frequencies of 48.7% and 15.3%, respectively. The highest and lowest antibiotic resistance rate was seen against azteroenam (94.2%) and amikacin (44.9%), respectively. Fluoroqinolone-resistant isolates were isolated with frequency of 45.8%. Multi-drug resistant (MDR) isolates were detected in 62.8% of isolates. The resistance rate in exoU (+) isolates was 86% compared to 66% in exoS (+) isolates. The high frequencies of virulence genes detected in our clinical isolates with notable antibiotic resistance rates indicate the potential risk of these isolates in nosocomial infections.

Different distribution of Helicobacter pylori EPIYA- cagA motifs and dupA genes in the upper gastrointestinal diseases and correlation with clinical outcomes in iranian patients.

AIM: Our aim was to determine the EPIYA-cagA Phosphorylation sites and dupA gene in H. pylori isolates among patients with upper gastrointestinal diseases. BACKGROUND: Pathogenicity of the cagA-positive Helicobacter pylori is associated with EPIYA motifs and higher number of EPIYA-C segments is a risk factor of gastric cancer, while duodenal ulcer-promoting gene (dupA) is determined as a protective factor against gastric cancer. PATIENTS AND METHODS: A total of 280 non-repeated gastric biopsies obtained from patients undergoing endoscopy from January 2013 till July 2013. Samples were cultured on selective horse blood agar and incubated in microaerophilic atmosphere. The isolated organisms were identified as H. pylori by Gram staining and positive oxidase, catalase, and urease tests. Various motif types of cagA and the prevalence of dupA were determined by PCR method. RESULTS: Out of 280 specimens, 128 (54.7%) isolated organisms were identified as H. pylori. Of 120 H. pylori isolates, 35.9% were dupA positive and 56.26% were cagA positive, while cagA with ABC and ABCC motifs were 55.5% and 44.5%, respectively. Fifty six percent of the isolates with the ABCC motif have had dupA genes. We also found a significant association between strains with genotypes of dupA-ABC and duodenal ulcer disease (p = 0.007). CONCLUSION: The results of this study showed that the prevalence of cagA-positive H. pylori in Shiraz was as high as in western countries and higher numbers of EPIYA-C segments were seen in gastric cancer patients. We may also use dupA as a prognostic and pathogenic marker for duodenal ulcer disease and cagA with the segment C for gastric cancer and gastric ulcer disease in this region.

The cutaneous bacterial microflora of the bodybuilders using anabolic-androgenic steroids.

BACKGROUND: Anabolic-androgenic steroids (AAS) abuse by the athletes has dramatically increased during the recent decades. These substances might increase the skin lipids and enhance the cutaneous microbial proliferation. OBJECTIVES: The current study aimed to investigate the potential side effects of AAS on the bacterial microflora colonization of the bodybuilders` skin. PATIENTS AND METHODS: The skin samples of 94 male bodybuilders (71 AAS users, 23 non-AAS users) and 46 subjects of the control group, with similar gender and age, were cultured and incubated in both aerobic condition to isolate Staphylococcus aureus and anaerobic condition for Propionibacterium acnes. The isolated bacteria were identified by standard microbiological techniques. RESULTS: The skin lesions were more frequent in the body builders than the controls. Moreover, statistically significant differences were also observed in skin lesions among the AAS users and the non-AAS user athletes. The prevalence of S. aureus and P. acnes in the athletes was higher than that of the control group. In addition, there was a significant difference in distribution of P. acnes between the bodybuilders who used AAS and those who did not. CONCLUSIONS: A higher number of bacterial flora was found in the bodybuilders particularly those using AAS in comparison to the controls, which might be due to the influence of these AAS on the skin microflora and transmission of the bacteria through the direct contact of the naked skin with the exercise instruments.