High frequency of vancomycin resistant Enterococcus faecalis in children: an alarming concern.

INTRODUCTION: Enterococcus spp. is considered as important etiological agents of nosocomial infections. However, a little is known about the epidemiology of vancomycin resistant Enterococcus faecalis (VREF). The aim of this study was to investigate the frequency of VREF and detecting of two prevalent resistance genes (vanA, vanB) at Children Medical Center Hospital, an Iranian referral pediatric Hospital. MATERIALS AND METHODS: During January 2013 to December 2013, 180 E. faecalis were isolated from clinical samples of hospitalized children. Antimicrobial testing was performed by Kirby-Bauer disk diffusion to gentamicin, amikacin, ceftriaxone, cefotaxime, ceftazidim, cefixime, piperacillin/tazobactam, cefepime, trimethoprim/sulfamethoxazole, erythromycin, clindamycin, linezolide and E-test method vancomycin and teicoplanin according to Clinical Laboratories Standards Institute (CLSI). Two prevalent resistance genes (vanA, vanB) were investigated in VREF isolates. RESULTS: Seventy-five (42%) of patients were male and 105 (58%) were female. Mean age of patients was 34.74 months. Cephalosporin resistance was found in majority of E. faecalis isolates (98.7 to ceftazidim, 95% to cefixime, 93.3% to ceftriaxone, and 89.4% to cefotaxime). Most of the isolated were susceptible to cefepime (91.7%). In addition, high level of erythromycin and clindamycin resistance was reported (93.4% and 91.2%). There were no linezolid-resistant E. faecalis among all isolates. Teicoplanin resistance was observed in 13.8% of E. faecalis (n = 25). Minimum Inhibitory concentration (MIC) ≥ 32 µg/ml for vancomycin was found in 29 isolates (16%) and vanA gene was detected in 21 (72%) VREF strains, while vanB gene was not detected in any of these isolates. The mortality rate of all cases was 3.4%. CONCLUSIONS: This study revealed high rate of vancomycin resistance in E. faecalis strains. Therefore, periodic surveillance of antibacterial susceptibilities is highly recommended to detect emerging resistance.

Impaired in-vitro responses to IL-12 and IFN-γ in Iranian patients with Mendelian susceptibility to mycobacterial disease

INTRODUCTION: Diagnosis of specific molecular defects of Mendelian susceptibility to mycobacterial diseases (MSMD) patients is important with respect to their clinical outcomes and their response to therapy. The aim of this study was to perform functional tests on blood samples of a group of patients who were suspected of having MSMD. METHODS: This study was performed on 11 cases who had mycobacterial infections and suspected MSMD. Whole blood cell culture was performed in presence of different stimulators. The supernatants were assayed for IFN-γ, IL-12p40 by ELISA method. RESULTS: All patients presented with complications of BCG vaccine in the form of localised lymphadenitis or disseminated BCG infection and chronic mycobacterial osteomyelitis. Infections with Salmonellaspecies occurred in two patients. In-vitro studies showed that 10 cases had impaired response to IL-12. However, the baseline levels of IL-12p40 were normal, while one of our patients may have a potential IFN-γ signalling defect or an IL-12p40 defect. CONCLUSIONS: Early detection of MSMD and commencing of appropriate combination therapy could prevent severe or even fatal complications of uncontrolled mycobacterial infections

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Impaired in-vitro responses to IL-12 and IFN-γ in Iranian patients with Mendelian susceptibility to mycobacterial disease.

INTRODUCTION: Diagnosis of specific molecular defects of Mendelian susceptibility to mycobacterial diseases (MSMD) patients is important with respect to their clinical outcomes and their response to therapy. The aim of this study was to perform functional tests on blood samples of a group of patients who were suspected of having MSMD. METHODS: This study was performed on 11 cases who had mycobacterial infections and suspected MSMD. Whole blood cell culture was performed in presence of different stimulators. The supernatants were assayed for IFN-γ, IL-12p40 by ELISA method. RESULTS: All patients presented with complications of BCG vaccine in the form of localised lymphadenitis or disseminated BCG infection and chronic mycobacterial osteomyelitis. Infections with Salmonella species occurred in two patients. In-vitro studies showed that 10 cases had impaired response to IL-12. However, the baseline levels of IL-12p40 were normal, while one of our patients may have a potential IFN-γ signalling defect or an IL-12p40 defect. CONCLUSIONS: Early detection of MSMD and commencing of appropriate combination therapy could prevent severe or even fatal complications of uncontrolled mycobacterial infections.

Candiduria in children: a first report from an Iranian referral pediatric hospital.

Candida spp. especially Candida albicans is considered as one of the most common cause of fungal infections. The aim of our study was to determine epidemiology of candiduria in children who were referred to an Iranian referral hospital. During May 2011 to February 2013, among 4813 urine culture positive, 209 candida spp. isolates (4.3%) was found. Forty-one percent of cadiduria infection was seen in patients between 1 month and 1 year, 24% in neonatant and 24% in patients 1 to 5 years. Cadiduria was mainly found in patients who had received more than 2 or 3 antibiotic during their hospitalization (37% and 24%, respectively). In our study, the highest frequency of cadiduria was seen in patients who had received more than 2 antibiotics and more than 3 antibiotics during their hospitalization; therefore, the strategic goals to optimize antimicrobial use including optimizing choice and duration of empiric therapy as well as monitoring and providing feedback regarding antibiotic resistance are recommended.

Diarrhoeagenic E. coli pathotypes in children with and without diarrhoea in an Iranian referral paediatrics centre.

Diarrhoeagenic Escherichia coli can be considered as the most important etiologic agents of diarrhoea in the Islamic Republic of Iran, particularly in children. This study determined the frequency of diarrhoeagenic E. coli isolates collected from children with acute diarrhoea (n = 50) and a control group (n = 50) at an Iranian referral paediatric centre during a 1-year period. Using multiplex PCR, diarrhoeagenic E. coli was identified in 90% ofthe case group and 20% of controls. Enterotoxigenic E coli was the most frequently identified pathotype in both groups (26% in cases; 10% in controls). Shiga toxin-producing E. coli was the second most isolated pathotype (17%), followed by enteroaggregative E. coli (12%). No enteroinvasive E. coli and enteropathogenic E. coli strains were recovered. More than 80% of isolates harboured the fimH gene. This high proportion of diarrhoeagenic E. coli and diversity of E. coli types highlights the need for enhanced surveillance of gastroenteritis agents in children in this country.

Staphylococcus aureus infections in children in an Iranian referral pediatric hospital.

INTRODUCTION: Staphylococcus aureus is associated with various infections ranging from skin and soft tissues such as surgical site infections and abscesses to lower respiratory tracts and bloodstream. The aim of this study was to evaluate underlying condition of patients with S. aureus infections in an Iranian referral pediatric Hospital. MATERIAL AND METHODS: Information was extracted retrospectively from the medical records of patients who were diagnosed with S. aureus infections. Data obtained about the study subjects included basic demographics, reason for admission, culture site, length of hospital stay, and methicillin susceptibility. RESULTS: The underlyning condition of of patients with S. aureus infection during November 2011 and March 2013 were included in the study. The most frequent diagnosis in patients with S. aureus infection was jaundice (12%), abscess (10%), cellulitis (10%), wound infection (8%), septic arthritis (7%) and sezeire (5%). Wound was the most common infection sites among all subjects 34/98 (35%) following by blood (20/98, 20%) as well as skin and soft tissue (19/98, 19%). The proportion of MRSA infections among all S. aureus isolates was 79% (77/98) during the study period. In addition, 58/74 (78%) met the definition of Hospital-Associated Methicillin-Resistant S. aureus (HA-MRSA) infections and the rest; 20/24 patients (83%), were classified as Community-Associated Methicillin-Resistant S. aureus (CA- MRSA). CONCLUSION: In our study, the high frequency of MRSA was found not only in HA S. aureus but also in CA S. aureus isolates; therefore, the strategic goals to optimize antimicrobial use including

Clonal spread of vancomycin resistance Enterococcus faecalis in an Iranian referral pediatrics center.

Vancomycin-resistant enterococci (VRE) represent as an immediate threat to public health. Since few active compounds are available for VRE infections, rapid identification of these isolates are essential. In the absence of any report on the genetic relatedness of Enterococcus faecalis especially Vancomycin-resistant E. faecalis (VREF) isolates in Iran, we undertook this study to characterize these isolates using random amplification of polymorphic DNA (RAPD-PCR) genotyping method. In this study, E. faecalis strains isolated from various samples collected from different wards of Children Medical Hospital (Tehran, Iran). These isolates were identified by standard laboratory procedures and tested for antimicrobial resistance to vancomycin and teicoplanin. The genetic similarity of the strains was investigated by amplification of the RAPD-PCR. In our study among 91 E. faecalis isolates, 15 (16%) were identified as VREF. The similarity pattern built for E. faecalis isolates by RAPD-PCR, demonstrated the presence of four distinct clusters (A, B, C, D). It is of interest to note that 100% of VREF isolates belonged to Clusters A, indicating that there may have occurred horizontal transmission of the same strain between patients. In conclusion, rapid spread of VREF from a clonal origin calls for implementation of careful isolation and infection control measures. Therefore, environmental control by routine disinfection of patient area as well as screening of high risk patients and isolation of colonized patients should be imposed in order to diminish risk of acquiring nosocomial VRE.

Pseudomonas aeruginosa infection among cystic fibrosis and ICU patients in the referral children medical hospital in Tehran, Iran.

INTRODUCTION: Pseudomonas aeruginosa is one of the important causes of hospital-acquired infections in Intensive Care Unit (ICU) and considered as a major determinant of morbidity and mortality in patients affected by cystic fibrosis (CF). The aim of this study was to investigate clonal diversity among randomly picked P. aeruginosa isolates of CF and the other hospitalized patients in ICU. METHODS: Cultivation, identification, and antimicrobial susceptibility testing of P. aeruginosa isolates were performed using standard techniques. The genetic similarity of the strains was investigated by amplification of the Enterobacterial Repetitive Intergenic Consensus-polymerase chain reaction (ERIC-PCR) sequence. RESULTS AND DISCUSSION: Among 49 isolates, sixteen were isolated from 11 patients affected by CF and 33 came from an epidemiological investigation of 25 P. aeruginosa infected patients of ICU. Five clusters were generated for all isolates analyzed through ERIC-PCR genotyping. Two major clusters (B and C) were discovered in P. aeruginosa isolates of ICU and CF patients during the whole period of this study. Fifteen unique antibiogram patterns obtained from all isolates and multi-resistant P. aeruginosa (MRPA) were identified in 23 isolates (47%). MRPA isolates were detected in all clusters (except A) while pan-resistant isolates were recovered only in cluster C. The high prevalence of related or identical isolates in CF and non-CF patients can be due to transmission of particular dominant clones in ICU ward. Therefore, enhanced infection-control may become necessary to prevent further spread of clonal strains.

Diarrhoeagenic E. coli pathotypes in children with and without diarrhoea in an Iranian referral paediatrics centre

Diarrhoeagenic Escherichia coli can be considered as the most important etiologic agents of diarrhoea in the Islamic Republic of Iran, particularly in children. This study determined the frequency of diarrhoeagenic E. coliisolates collected from children with acute diarrhoea [n= 50] and a control group [n= 50] at an Iranian referral paediatric centre during a 1-year period. Using multiplex PCR, diarrhoeagenic E. coli was identified in 90% of the case group and 20% of controls. Enterotoxigenic E. coli was the most frequently identified pathotype in both groups [26% in cases; 10% in controls]. Shiga toxin-producing E. coli was the second most isolated pathotype [17%], followed by enteroaggregative E. coli [12%]. No enteroinvasive E. coliand enteropathogenic E. colistrains were recovered. More than 80% of isolates harboured the fimHgene. This high proportion of diarrhoeagenic E. coli and diversity of E. coli types highlights the need for enhanced surveillance of gastroenteritis agents in children in this country

Genotypic characteristics of Pseudomonas aeruginosa strains circulating in the tertiary referral Children's Medical Hospital in Tehran, Iran.

Pseudomonas aeruginosa is an important pathogen with the ability to cause infection in all departments of the hospital, especially in intensive care units (ICUs). The aim of this study is to analyse the epidemiological relationships among clinical P. aeruginosa strains isolated from different wards of the Children's Medical Center Hospital (Tehran, Iran). These isolates were identified by standard laboratory procedures and tested for antimicrobial resistance to several antibiotic agents. The genetic similarity of the strains was investigated by amplification of the enterobacterial repetitive intergenic consensus sequence (ERIC-PCR). During the study period, 87 non-duplicate patients were colonised or infected with P. aeruginosa. Among the isolates, resistance to piperacillin/tazobactam was low (27%), followed by amikacin (31%), gentamicin (33%), imipenem (33%), ciprofloxacin (36%) and meropenem (39%). Thirty-five patients (40.2%) were either colonised or infected with a multidrug-resistant P. aeruginosa strain (MDRP) over a one-year period, and 17 isolates were non-susceptible to all the tested antibiotics. One predominant profile (D) was identified in 59 strains. This profile first appeared in the paediatric intensive care unit (PICU) and infection ward in June 2010, and circulated around all wards up to the end of the study period. Of the 35 MDRP, 22 (62.8%) were found to be profile D. Molecular typing of the isolates suggests considerable cross-transmission of P. aeruginosa not only between patients in one ward but also between patients from different wards. This can be explained partly by the high number of patients transferred between different wards of the hospital.

Multiple organ involvement with hydatid cysts.

Hydatid disease is the most common infections worldwide, but it rarely involves multiple organs. Herein, a 12-year-old boy is presented, who was admitted to Children's Medical Center, Tehran University of Medical Sciences, Tehran, Iran with symptoms of irritability, sleepless, and weakness of the extremities. Patient's brain computed tomography (CT) scan with contrast media showed large multilocular cystic lesions in right temporal lobe associated with two other smaller similar cystic lesions in centrum semiovale bilaterally. Abdominal sonography revealed intestinal mesenteric and a cardiac cyst. Abdomino-pelvic CT scan showed a cyst medial to the cecum and a cortical cyst in the left kidney as well as a heart cyst. The echocardiography confirmed hydatid cysts at apical and interventricular septum. Serology test was positive for hydatid cyst. Albendazole and praziquantel were started for the patient immediately and right temporal lobe lesions were removed via neurosurgery intervention. After one month, cardiac and mesenteric cysts were operated during two separate surgeries. Pathologic findings of all cysts were compatible with hydatid cyst. Cystic hydatidosis should be suspected in any cystic mass, whilst prompt diagnosis and appropriate treatments are the keys in management of affected patients.

Neurite outgrowth of dorsal root ganglia is delayed and arrested by aspirin.

In this study, the effect of increasing doses of aspirin on the neurite outgrowth of Dorsal Root Ganglia (DRG) was investigated. DRG were cultured in complete medium (DMEM + 10% FCS +100 ng/ml NGF + collagen Type1 in substratum in 96 multiwell plate) in the presence of concentration of 1.25, 2.5, 5 and 10 mM aspirin. The neurite outgrowth of DRG was followed in comparison with controls that lack aspirin. 10 mM aspirin treated DRG showed delayed neurite outgrowth and after 7 days it reached the same DRG neurite outgrowth control wells after 18 hrs. This growth has delayed approximately one week and showed no further development and in such stage the cells became apoptos. However at concentrations of 1.25, 2.5, 5 mM of aspirin, outgrowth was observed after 18-24 hrs. Although the rate of growth was lower than control, it was not significant. In the other experiment, when DRG cultured for one week in complete medium then treated with aspirin, at 10 mM, DRG neurite outgrowth was stopped, while it was continued in the control. It seem that the aspirin affected DRG became apoptosis.

Genotoxic risk assessment of drinking water consumed in the city of Tehran, Iran.

Two hundred liters of drinking water of the city of Tehran were collected as tap water in a glass container and passed through a XAD-2 resin column at a flow rate of 60-100 ml/minute. The adsorbed materials were eluted with acetone, dried and dissolved in 2 ml dimethylsulfoxide (DMSO). Doses up to 10 microliters did not elevate sister chromatid exchanges (SCE) in CHO fibroblasts, while 20 microliters caused a significant increase in SCE. However, this dose did not show any chromosomal aberration (CA) in V79 fibroblasts. Doses up to 50 microliters of the extract did not increase his+ spontaneous revertant colonies in Salmonella typhimurium tester strain TA98, TA100 and TA102 in the absence of S9 metabolizing mixture, neither induced DNA alkaline labile sites in V79 fibroblasts.