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Cells with osteogenic potential are believed to be an ideal source for bone tissue bioengineering. Large bone defects require temporary substitution of the damaged parts. In this respect, the transplantation of bone cells cultured on osteogenic substrates has been investigated. To use the natural bone matrix, one approach is the so-called demineralized bone matrix (DBM). In this study, we evaluated the interaction of human fetal osteoblasts (hFOB 1.19 cells, a human fetal osteoblastic cell line) with DBM fragments. No additional bone differentiation inducer was used other than the DBM itself. The samples were processed, had adhesion pattern evaluated and analyzed by light microscopy (cytochemical and immunocytochemical analysis) and electron microscopy (scanning and transmission). The adhesion pattern of hFOB cells on DBM was similar to what was observed on the cell culture plate. Morphological analysis showed that the hFOB cells had emitted filopodia and cellular projections on both controls and DBM. On DBM, the adhered cells emitted prolongations and migrated into the matrix. The monolayer growth pattern was observed as well as the accumulation of filamentous and reticulate extracellular materials when hFOB cells were cultured on the DBM surface. EDS analysis revealed the deposition of calcium on DBM. Immunocytochemical data showed that the hFOB cells were able to secrete extracellular matrix molecules such as fibronectin and laminin on DBM. Our data indicate that DBM successfully stimulates the osteoblastic phenotype of osteoblast-like cells and corroborate with the fact that DBM is a considerable natural matrix that promotes fractured-bone healing.

A rising incidence of extended-spectrum beta-lactamase-producing Enterobacteriacae has caused an increased use of carbapenems and subsequent spread of carbapenem-resistant strains. Two recent artic...

Due to the increasing demand for a bone marrow study model, we developed a natural scaffold from decellularized bovine bone marrow (DeBM). The obtained bioscaffold was analyzed after the decellularization process; histological staining, scanning and transmission electron microscopy confirmed the preservation of its native 3D-architecture; including blood vessels and cell niches as well as the integrity of important components of the extracellular matrix; Collagen III, IV and fibronectin. In addition to biochemical composition, physical properties of the bone marrow were also conserved. We evaluated the suitability of this bio-scaffold as a tridimensional culture platform. Seeding experiments with umbilical cord-derived hematopoietic stem cells and human bone marrow stromal cell line HS5 demonstrated that this scaffold is capable of supporting hematopoietic and stromal cell adhesion and proliferation without the need of exogenous factors. DeBM provided an inductive environment for the repopulation of the bone marrow inducing the expression of SDF-1, HGF and SCF by seeded stromal cells. The presence of these potent hematopoietic chemoattractants would be crucial for ex vivo long-term culture of HSCs, and for recreating the natural microenvironment of the bone marrow for bioengineering applications. We conclude that the decellularization process succeeded in preserving the 3D structure and mechanical properties of the bone marrow. The resulting scaffold is suitable for cell culture, representing an advantageous bone marrow experimental model, and potentially an effective platform for CD34+ HSC expansion and differentiation for clinical applications.

Introduction. Bloodstream infection is one of the most frequent and challenging hospital-acquired infections and it is associated with high morbidity, mortality and additional use of healthcare resources. Hypothesis/Gap Statement: Bloodstream infections have consequences for the patient, such as the evolution to mortality and inappropriate empirical antibiotic prescription, especially when caused by multidrug-resistant Gram-negative bacilli. Objective. To assess the impact of bloodstream infection and the status of multidrug resistance (MDR) in the evolution of patients who received inappropriate initial antibiotic therapy. Methods. A retrospective surveillance was conducted on nosocomial bloodstream infections caused by Gram-negative bacilli (GNB) from January 2012 to December 2018 in an adult intensive care unit of a Brazilian tertiary teaching hospital. Results. We identified 270 patients with GNB nosocomial bacteremia. Non-survivors were older (with an average age of 58.8 years vs 46.9 years, P=P=P=P=0.0013) than survivors. In a logistic regression model, inappropriate empirical antibiotic therapy was not an independent predictor of mortality, different from mechanical ventilator (P=P=0.0051; OR=2.5; 95% CI=1.3–4.9) and immunosuppression (P=0.0066; OR=2.6; 95% CI=1.3–5.2). In contrast, in a separate model, MDR was strongly associated with the prescription of inappropriate initial antibiotic therapy (P=0.0030; OR=5.3; 95% CI=1.7–16.1). The main isolated pathogens were Acinetobacter baumannii (23.6 %) and Klebsiella pneumoniae (18.7 %). The frequency of MDR organisms was high (63.7 %), especially among non-fermenting bacilli (60.9 %), highlighting A. baumannii (81.6 %) and Pseudomonas aeruginosa (41.8 %). Conclusion. Illness severity (septic shock and immunosuppression) and mechanical ventilation were identified as predictors of mortality. Additionally, MDR was a major determinant of inappropriate antibiotic empirical therapy, but not associated with mortality, and both characteristics were not statistically associated with death.

It appears inevitable that severe acute respiratory syndrome coronavirus 2 will continue to spread. Although we still have limited information on the epidemiology of this virus, there have been multiple reports of superspreading events (SSEs), which are associated with both explosive growth early in an outbreak and sustained transmission in later stages. Although SSEs appear to be difficult to predict and therefore difficult to prevent, core public health actions can prevent and reduce the number and impact of SSEs. To prevent and control of SSEs, speed is essential. Prevention and mitigation of SSEs depends, first and foremost, on quickly recognizing and understanding these events, particularly within healthcare settings. Better understanding transmission dynamics associated with SSEs, identifying and mitigating high-risk settings, strict adherence to healthcare infection prevention and control measures, and timely implementation of nonpharmaceutical interventions can help prevent and control severe acute respiratory syndrome coronavirus 2, as well as future infectious disease outbreaks.

Carbapenemase-producing organisms are pandemic and a significant threat to public health. We investigated the clonal relatedness of colistin-resistant Klebsiella pneumoniae strains producing KPC-type carbapenemase (KPC-KP) causing subsequent infections or colonization. Moreover, we aimed to gain insight into the ability of biofilm production in K. pneumoniae strains producing carbapenemase. Twenty-two consecutive KPC-KP and one KPC-negative strain was identified from an adult intensive care unit in Brazil. Seventy-five percent of isolates that harbored the blaKPC gene exhibited genetic relatedness by pulsed-field gel electrophoresis, and none presented the plasmid-mediated mcr-1 and blaNDM genes. This study showed that the majority of repeated KPC infections in adults were caused by a clone that caused the previous infections/colonizations even after a long period of time and illustrates the capacity of multiple clones producing biofilms to coexist in the same patient at the same time, becoming a reservoir of KPC-KP in the hospital environment.

Sorafenib has been widely used to treat unresectable hepatocellular carcinoma (HCC) but most studies have been done in Child-Pugh A (CP-A) patients with well-preserved liver function. We evaluated the overall survival (OS) and tolerance to sorafenib in a large cohort of Child-Pugh B (CP-B) HCC patients as compared to CP-A HCC patients. We prospectively studied 130 patients with advanced HCC who started sorafenib between January 2011 and December 2015. Patients were classified as CP-A (n = 65) or CP-B (n = 65). The average OS for all 130 patients was 10 months. CP-A patients had a median survival rate significantly longer than CP-B patients: 12 months vs. 6 months. The OS found in our group of CP-B patients was 6.5 months, which is higher than that found in most studies done so far. When stratified, our CP-B patients had better OS than ever reported. The dose of the drug was interrupted due to adverse events (AEs) in 38 (29%) of the patients, of whom 20 (30%) were CP-A patients and 18 (28%) were CP-B patients. This real-life cohort of CP-B HCC patients treated with sorafenib had a higher survival than that described in the literature, with a satisfactory safety profile. Despite the high prevalence of severe AEs in CP-B patients, there were fewer treatment interruptions in this group, indicating that Child-Pugh B patients can tolerate treatment and may benefit from sorafenib.

In this study, we describe the frequency of virulence genes in Klebsiella pneumoniae carbapenemase-2-producing Klebsiella pneumoniae (KPC-KP), including hypervirulent (hv) and hypermucoviscous (hm) strains by whole-genome sequencing. We also evaluate the capacity for biofilm formation by using phenotypic techniques. The occurrence of several virulence genes (fimABCDEFGHIK, mrkABCDFHJ, ecpA, wabG, entB, ugE, irp1, irp2, traT, iutA and ureADE) and a high frequency of hvhmKPC-KP isolates was found. Most hospital-associated lineages of KPC-KP belong to the international clonal group 258 (CG258). Biofilm formation was a constant feature among 90.9 % of KPC-KP strains. This report suggests a close relationship between ST437 and weak biofilm production, given that all weakly biofilm-producing strains belonged to this sequence type. This also supports the dissemination of KPC-KP containing numerous virulence determinants belonging to the biofilm-producing CG258 type in Brazil, including hv and hm strains. These factors allow this pathogen to cause infections, leading to its rapid expansion and persistence in hospital settings.

The purpose of this study was to identify the risk factors that predispose patients who are hospitalized with pressure ulcers (PUs) colonized by Gram-negative bacilli (GNB) to develop bacteremia. In addition, we also detected main phenotypes of resistance in infected and uninfected PUs. A prospective cohort study was conducted at the Clinical Hospital of the Federal University of Uberlândia including patients with Stage II or greater PUs, colonized or not with GNB, from August 2009 to July 2010. Infected ulcers were defined based on clinical signs and on positive evaluation of smears of wound material translated by a ratio of polymorphonuclear cells to epithelial cells ≥2:1, after Giemsa staining. A total of 60 patients with Stage II PUs were included. Of these 83.3% had PUs colonized and/or infected. The frequency of polymicrobial colonization was 74%. Enterobacteriaceae and GNB non-fermenting bacteria were the most frequent isolates of PUs with 44.0% of multiresistant isolates. Among patients who had infected PUs, six developed bacteremia by the same microorganism with a 100% mortality rate. In addition, PUs in hospitalized patients were major reservoir of multiresistant GNB, also a high-risk population for the development of bacteremia with high mortality rates. Keywords: Pressure ulcer, Gram-negative bacilli, Colonization, Bacteremia

Dear editor, Over the years, became worryingly the overuse and misuse of antimicrobials to the hospital setting, particularly in lower and middle-income countries (LMICs) where antibiotic utilizati...

This study used whole-genome sequencing (WGS) and PFGE to analysis KPC-2-producing Klebsiella pneumoniae strains from clinical specimens collected in Brazilian hospitals. The study identifies the emergence of a novel small IncX3 plasmid (pKPB11), 12,757-bp in length, in a high-risk K. pneumoniae ST11/CG258 lineage, a successful clonal group in Brazil, carrying the bla

The dissemination of antimicrobial resistance genes and the bacterium that harbor them have increasingly become a public concern, especially in low- and middle-income countries. The present study used whole-genome sequencing to analyze 10 KPC-2-producing

Background Carbapenem-resistant Acinetobacter baumannii (CR-Ab) has become a worrying health care problem, mainly in developing countries, such as Brazil. The objective was to investigate the prevalence and prognostic factors for CR-Ab infections at a Brazilian university hospital and examine the impact of inappropriate antimicrobial therapy on patient outcome. Methods A retrospective study on hospitalized patients with CR-Ab infections was carried out from January 2013 to December 2017. An epidemiologic analysis was carried out to determine the frequency of infections, the epidemiologic indicators by year, the risk factors for 30-day mortality, and the impact of inappropriate therapy. Results A total of 489 patients were included in the study. A rate of 0.7 per 1,000 patient-day CR-Ab infections was observed, mostly in the lungs (54.7%), and predominantly in the adult intensive care unit. The occurrence of infections by CR-Ab per 1,000 patient-days in November 2014 exceeded the established control limit, confirming an outbreak. Conclusions The prevalence of CR-Ab increased in the investigated hospital, passing to an endemic pathogen with a direct impact on mortality and the control of these strains.

INTRODUCTION: The present study aimed to determine the incidence of health care-associated infections (HCAIs) and identify the main resistant microorganisms in intensive care unit (ICU) patients in a Brazilian university hospital. METHODS: A retrospective cohort study was conducted in a Brazilian teaching hospital between 2012 and 2014. RESULTS: Overall, 81.2% of the infections were acquired in the ICU. The most common resistant pathogenic phenotypes in all-site and bloodstream infections were oxacillin-resistant coagulase-negative staphylococci and carbapenem-resistant Acinetobacter spp. (89.9% and 87.4%; 80.6% and 70.0%), respectively. CONCLUSIONS: There is an urgent need to focus on HCAIs in ICUs in Brazil.

A great investment has been done in the last decades in the development of numerical and qualitative assessment methods to classify the ecological quality of water bodies. Yet, in spite of all attempts to avoid subjectivity, expert judgment is still used at numerous steps of the ecological classification and is considered by some authors as indispensible for management purposes. Thus, the aim of this study is to test the hypothesis that expert judgment, when done by the adequate experts (limnologists/river ecologist) with experience in the study area (i.e., natural conditions and expected communities), could be as good as quantitative indices and measures (i.e., result in the same classification), but quicker and with lower cost. For that we compared the classifications (on 13 aspects of rivers ecosystems) attributed by two experts to 20 sites (10 each) located in their study areas, with the classifications of ecological quality based on biological indices (for invertebrates and diatoms), hydromorphology and water chemistry, calculated by an independent team. Our results show that assessments made by experts and those calculated through indices (biological quality and hydromorphology) are globally very similar (RELATE test; Rho = 0.442; p

Made available in DSpace on 2018-11-26T17:55:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Background: Minimal structure is required for effective prevention of health careeassociated infection (HAI). The objective of this study was to evaluate the structure for prevention of HAI in a sample of Brazilian hospitals. Methods: This was a cross-sectional study from hospitals in 5 Brazilian regions (n = 153; total beds: 13,983) classified according to the number of beds; 11 university hospitals were used as reference for comparison. Trained nurses carried out the evaluation by using structured forms previously validated. The evaluation of conformity index (CI) included elements of structure of the Health CareeAssociated Prevention and Control Committee (HAIPCC), hand hygiene, sterilization, and laboratory of microbiology. Results: The median CI for the HAIPCC varied from 0.55-0.94 among hospital categories. Hospitals with > 200 beds had the worst ratio of beds to sinks (3.9; P

Transcriptional regulation is crucial for all biological processes and well investigated at the molecular level for a wide range of organisms. However, it is quite unclear how innovations, such as the activity of a novel regulatory element, evolve. In the case of transcription factor (TF) binding, both a novel TF and a novel-binding site would need to evolve concertedly. Since promiscuous functions have recently been identified as important intermediate steps in creating novel specific functions in many areas such as enzyme evolution and protein–protein interactions, we ask here how promiscuous binding of TFs to TF-binding sites (TFBSs) affects the robustness and evolvability of this tightly regulated system. Specifically, we investigate the binding behavior of several hundred TFs from different species at unprecedented breadth. Our results illustrate multiple aspects of TF-binding interactions, ranging from correlations between the strength of the interaction bond and specificity, to preferences regarding TFBS nucleotide composition in relation to both domains and binding specificity. We identified a subset of high A/T binding motifs. Motifs in this subset had many functionally neutral one-error mutants, and were bound by multiple different binding domains. Our results indicate that, especially for some TF–TFBS associations, low binding specificity confers high degrees of evolvability, that is that few mutations facilitate rapid changes in transcriptional regulation, in particular for large and old TF families. In this study we identify binding motifs exhibiting behavior indicating high evolutionary potential for innovations in transcriptional regulation.

Summary Background Healthcare-associated infection (HAI) represents a major problem for patient safety worldwide. Aim To provide an up-to-date picture of the extent, aetiology, risk factors and patterns of infections in intensive care units (ICUs) in 28 Brazilian hospitals of different sizes. Methods A one-day point prevalence survey in 2016 enrolled the ICUs of hospitals from the 12 meso-regions in Minas Gerais state, south-east Brazil. Hospitals were classified as university or non-university hospitals. All patients with >48 h of admission to the study ICUs at the time of the survey were included. Findings In total, 303 patients were studied; of these, 155 (51.2%) were infected and 123 (79.4%) had at least one ICU-acquired infection. The most common ICU-acquired infections were pneumonia (53.0%) and bloodstream infection (27.6%). One hundred and nineteen bacterial isolates were cultured; the most common were Acinetobacter baumannii (27.1%), Pseudomonas aeruginosa (27.1%) and Staphylococcus aureus (39.0%). According to type of infection, the most common pathogens were P. aeruginosa (30.4%) in pneumonia, coagulase-negative staphylococci (23.4%) and Enterobacteriaceae (23.4%) in bloodstream infections, and Enterobacteriaceae (47.6%) in urinary tract infections. Conclusion This study found that the overall prevalence of ICU-acquired infections in surveyed Brazilian hospitals was higher than that reported in most European countries and the USA. A greater proportion of infections were caused by non-fermenting Gram-negative bacteria. These observations, along with a high rate of antimicrobial use, illustrate the urgent need for HAIs to be a priority in the public health agenda of Brazil.

Background: In Brazil, ventilator-associated pneumonia (VAP) caused by carbapenem resistant Acinetobacter baumannii and Pseudomonas aeruginosa isolates are associated with significant mortality, morbidity and costs. Studies on the clonal relatedness of these isolates could lay the foundation for effective infection prevention and control programs. Objectives: We sought to study the epidemiological and molecular characteristics of A. baumannii vs. P. aeruginosa VAP in an adult intensive care unit (ICU). Methods: It was conducted a cohort study of patients with VAP caused by carbapenem resistant A. baumannii and P. aeruginosa during 14 months in an adult ICU. Genomic studies were used to investigate the clonal relatedness of carbapenem resistant OXA-23-producing A. baumannii and P. aeruginosa clinical isolates. The risk factors for acquisition of VAP were also evaluated. Clinical isolates were collected for analysis as were samples from the environment and were typed using pulsed field gel electrophoresis. Results: Multivariate logistic regression analysis identified trauma diagnosed at admission and inappropriate antimicrobial therapy as independent variables associated with the development of A. baumannii VAP and hemodialysis as independent variable associated with P. aeruginosa VAP. All carbapenem resistant clinical and environmental isolates of A. baumannii were OXA-23 producers. No MBL-producer P. aeruginosa was detected. Molecular typing revealed a polyclonal pattern; however, clone A (clinical) and H (surface) were the most frequent among isolates of A. baumannii tested, with a greater pattern of resistance than other isolates. In P. aeruginosa the most frequent clone I was multi-sensitive. Conclusion: These findings suggest the requirement of constant monitoring of these microorganisms in order to control the spread of these clones in the hospital environment. Keywords: Acinetobacter baumannii, Intensive care unit, Molecular epidemiology, Multidrug-resistant

The rates of multidrug-resistant, extensively drug-resistant and pandrug-resistant isolates amongst non-fermenting Gram-negative bacilli, particularlyPseudomonas aeruginosa, have risen worldwide. The clinical consequence of resistance and the impact of adverse treatment on the outcome of patients withP. aeruginosabacteraemia remain unclear. To better understand the predictors of mortality, the clinical consequence of resistance and the impact of inappropriate therapy on patient outcomes, we analysed the first episode ofP. aeruginosabacteraemia in patients from a Brazilian tertiary-care hospital during the period from May 2009 to August 2011. Antimicrobial susceptibility testing was conducted; phenotypic detection of metallo-β-lactamase (MBL) and PCR of MBL genes were performed on carbapenem-resistant strains. Amongst the 120P. aeruginosaisolates, 45.8 % were resistant to carbapenem and 36 strains were tested for MBL detection. A total of 30 % were phenotypically positive and, of these, 77.8 % expressed an MBL gene,blaSPM-1(57 %) andblaVIM-type (43 %). The resistance rates to ceftazidime, cefepime, piperacillin/tazobactam, carbapenem, fluoroquinolone and aminoglycoside were 55, 42.5, 35, 45.8, 44 and 44 %, respectively. Previous antibiotic use, length of a hospital stay ≥30 days prior toP. aeruginosa, haemodialysis, tracheostomy, pulmonary source of bacteraemia and Intensive Care Unit admission were common independent risk factors for antimicrobial resistance. Cefepime resistance, multidrug resistance and extensive drug resistance were independently associated with inappropriate therapy, which was an important predictor of mortality, being synergistic with the severity of the underlying disease.

The assessment of risk factors for the nosocomial acquisition of colonization and infection by vancomycin-resistant Enterococcus faecium (VREfm) is often problematic due to scarce data on antibiotic use. A 30-month prospective cohort study was conducted to characterize VREfm strains isolated during an outbreak and endemic period, identifying the risk factors, antibiotic consumption, and prevalence of virulence determinants.The study was conducted in a tertiary care hospital. A representative number (171 patients) of isolates that were classified as resistant to high-level vancomycin (minimum inhibitory concentration (MIC) ≥ 256 μg/ml) were investigated.Among 171 colonized patients, 22 (12.9%) developed VRE infection. All VREfm isolates harboured vanA genes. Genes codifying virulence factors such as enterococcal surface protein (esp), aggregation substance 1 (asa1), and gelatinase (gelE) were detected in the VREfm studied. All patients infected with VRE had previously been colonized and became infected on average 14 days after colonization. Only previous use of aminoglycosides was a risk factor independently associated with VRE infection; however, glycopeptide consumption in defined daily doses (DDD) per 1000 patient-days was associated with the presence of this microorganism. The monthly colonization pressure ranged from 0.004% to 1.32% during the 30-month study period.We found a high incidence of VRE in a tertiary care hospital, independently associated with the prior use of aminoglycosides and the administration of glycopeptides.

INTRODUCTION:The frequency of methicillin-resistant Staphylococcus aureus (MRSA) has increased in the community. This study evaluated the prevalence of MRSA and community-acquired (CA)-MRSA in 120 healthy elderly.METHODS:The MRSA were evaluated for the presence of the IS256, mecA, agr, icaA, icaD, fnbB , and pvl genes with PCR. Results: Frequency of S. aureus and MRSA colonization was 17.8% and 19%, respectively. CA-MRSA isolate showed SCC mec IV, fnbB+ , and icaD+ .CONCLUSIONS:CA-MRSA was detected, with genotype determined as SCC mec type IV/IS256/ fnbB+ / icaA / icaD+ / bbp-/agr2 / bap / pvl, characterizing this population as a possible reservoir of this organism in the community.

Summary Background Carbapenem-resistant Enterobacteriaceae (CRE), especially those that produce Klebsiella pneumoniae carbapenemase (KPC) and are associated with colistin resistance, pose a severe health threat due to the limited treatment options. Aim To describe two outbreaks of KPC-producing K. pneumoniae in an adult intensive care unit (AICU) in Brazil. In May 2015, 14 patients had colistin-susceptible KPC-producing strains (ColS-KPC), and in July 2015, nine patients had colistin-resistant KPC-producing strains (ColR-KPC). Methods Between September 2014 and August 2015, we performed surveillance at a university hospital and all CRE were tested for bla KPC genes. Clonality was investigated by pulsed-field gel electrophoresis. Resistance to colistin was confirmed by broth microdilution method. Consumption of carbapenems and colistin was expressed as defined daily doses. Findings In all, 111 patients with CRE were identified during the surveillance period; K. pneumoniae was the major isolate (77.13%). The two outbreaks were identified when infection rates (KPC per 1000 patient-days) exceeded the background level. Rates of carbapenem and colistin consumption were high. Control measures (bedside alcohol gel, contact precautions, regular rectal swabs) did not curtail the outbreaks. Mortality rates were 42.9% and 44.4% for ColS-KPC- and ColR-KPC-infected patients, respectively. After the death of four infected patients with ColR-KPC, the unit was closed to new admissions. Conclusion Our experience demonstrates the serious risks presented by KPC, and especially ColR-KPC, in Brazilian AICUs. Selective pressure from excessive antibiotic use and transmission on healthcare workers' hands were likely the major factors in transmission.

We described a comprehensive analysis of the molecular epidemiology of multidrug-resistant (MDR) P. aeruginosa. Molecular analysis included typing by Pulsed Field Gel Electrophoresis, identification of genes of interest through PCR-based assays and sequencing of target genes. Case-control study was conducted to better understand the prognostic of patients and the impact of inappropriate therapy in patients with bacteremia, as well as the risk factors of MDR infections. We observed a high rate of MDR isolates (40.7%), and 51.0% of them was independently associated with inappropriate antibiotic therapy. Bacteremia was detected in 66.9% of patients, and prolonged hospital stay was expressive in those resistant to fluoroquinolone. Plasmid-mediated quinolone resistance genes (PMQR), qnrS1 and aac(6’)Ib-cr, were detected in two different nosocomial isolates (5.3%), and the aac(6’)-Ib7 variant was detected at a high frequency (87.5%) in those negative to PMQR. The presence of mutations in gyrA and parC genes was observed in 100% and 85% of selected isolates, respectively. Isolates harboring PMQR genes or mutations in gyrA and parC were not closely related, except in those containing SPM (Sao Paulo metallo-β-lactamase) clone. In addition, there is no study published in Brazil to date reporting the presence of Pseudomonas aeruginosa isolates harboring both qnrS1 and aac(6’)Ib-cr genes, with alarming frequency of patients with inappropriate therapy.

Multidrug-resistant (MDR) Acinetobacter baumannii (Acb) is a rapidly emerging pathogen in healthcare settings. The aim of this study was to evaluate the predictors of poor outcome in patients with MDR Acb. This is the first report documenting factors influencing survival in patients with MDR Acb in this tertiary hospital. This study is a prospective of the hospital epidemiology database. A total of 73 patients with 84 Acb isolates were obtained between August 2009 and October 2010 in this hospital. In the present study, the 30-day mortality rate was 39.7%. Of 84 Acb isolates, 50 (59%) were MDR, nine (11%) were pan-resistant, and 25 (30%) were non-MDR. The non-MDR isolates were used as the control group. The factors significantly associated with multidrug resistance included previous surgeries, presence of comorbidity (renal disease), use of more than two devices, parenteral nutrition, and inappropriate antimicrobial therapy. Significant predictors of 30-day mortality in the univariate analysis included pneumonia, diabetes mellitus, renal disease, use of more than two devices, and inappropriate antimicrobial therapy administered within two days of the onset of infection. The factors associated with mortality in patients with MDR Acb infection in this study were: age > 60 years, pneumonia, diabetes mellitus, renal disease, use of more than two invasive procedures, and inappropriate antimicrobial therapy. Vigilance is needed to prevent outbreaks of this opportunistic and deadly pathogen.

Vancomycin-resistant enterococci (VRE) are important hospital pathogens and have become increasingly common in patients admitted to the intensive care unit (ICU). To determine the incidence and the risk factors associated with VRE colonisation among ICU patients, active surveillance cultures for VRE faecal carriages were carried out in patients admitted to the ICU of the University Hospital of Uberlândia, Minas Gerais, Brazil. Risk factors were assessed using a case-control study. Seventy-seven patients (23.1%) were found to be colonised with vanC VRE and only one patient (0.3%) was colonised with vanA VRE. Independent risk factors for VRE colonisation included nephropathy [odds ratio (OR) = 13.6, p < 0.001], prior antibiotic use (OR = 5.5, p < 0.03) and carbapenem use (OR = 17.3, p < 0.001). Our results showed a higher frequency (23.1%) of Enterococcus gallinarum and Enterococcus casseliflavus, species that are intrinsically resistant to low levels of vancomycin (vanC), without an associated infection, associated with prior antibiotic use, carbapenem use and nephropathy as comorbidity. This study is the first to demonstrate the risk factors associated with vanC VRE colonisation in ICU hospitalised patients. Although vanA and vanB enterococci are of great importance, the epidemiology of vanC VRE needs to be better understood. Even though the clinical relevance of vanC VRE is uncertain, these species are opportunistic pathogens and vanC VRE-colonised patients are a potential epidemiologic reservoir of resistance genes.

INTRODUCTION: Catheter-associated bloodstream infection (CA-BSI) is the most common nosocomial infection in neonatal intensive care units. There is evidence that care bundles to reduce CA-BSI are effective in the adult literature. The aim of this study was to reduce CA-BSI in a Brazilian neonatal intensive care unit by means of a care bundle including few strategies or procedures of prevention and control of these infections. METHODS: An intervention designed to reduce CA-BSI with five evidence-based procedures was conducted. RESULTS: A total of sixty-seven (26.7%) CA-BSIs were observed. There were 46 (32%) episodes of culture-proven sepsis in group preintervention (24.1 per 1,000 catheter days [CVC days]). Neonates in the group after implementation of the intervention had 21 (19.6%) episodes of CA-BSI (14.9 per 1,000 CVC days). The incidence of CA-BSI decreased significantly after the intervention from the group preintervention and postintervention (32% to 19.6%, 24.1 per 1,000 CVC days to 14.9 per 1,000 CVC days, p=0.04). In the multiple logistic regression analysis, the use of more than 3 antibiotics and length of stay >8 days were independent risk factors for BSI. CONCLUSIONS: A stepwise introduction of evidence-based intervention and intensive and continuous education of all healthcare workers are effective in reducing CA-BSI. INTRODUÇÃO: As infecções de corrente sanguínea associadas ao cateter (ICS-AC) são as infecções hospitalares mais frequentes em unidades de terapia intensiva neonatais. O objetivo do nosso estudo foi reduzir as ICS-AC através de um pacote de medidas de cuidados incluindo algumas estratégias de controle e prevenção destas infecções. MÉTODOS: Foi realizada uma intervenção desenhada para reduzir as infecções de corrente sanguínea associadas ao cateter com cinco procedimentos de base. RESULTADOS: Um total de 67 (26,7%) ICS-AC foi observado. Houve 46 (32%) episódios de sepse com critério microbiológico (24,1 por 1.000 catater dias(CVC-dias). Os neonatos do grupo, após a implementação da intervenção, tiveram 21 (19,6%) episódios de ICS-AC (14,9 por 1.000 CVC-dias). A incidência de ICS-AC reduziu significantemente entre os grupos pré-intervenção e pós-intervenção após a implementação das medidas (32% para 19.6%, 24.1 por 1.000 CVC-dias para 14.9 por 1.000 CVC-dias, p=0.04). Na análise de regressão logística múltipla, o uso de > três antibióticos e tempo de hospitalização > 8 dias foram fatores de risco independentes para ICS. CONCLUSÕES: A introdução de uma intervenção baseada em evidências e a educação intensiva e continuada de todos os profissionais de saúde são efetivas para a redução de ICS-AC.

This study evaluated the relationship between previous colonization of the oropharynx and development of ventilator-associated pneumonia through the classification of genomic fingerprint pattern by pulsed-field gel electrophoresis of both oxacillin-resistant and oxacillin-susceptible Staphylococcus aureus isolates obtained from hospitalized patients in an intensive care unit.

INTRODUCTION: Report the incidence of nosocomial infections, causative microorganisms, risk factors associated with and antimicrobial susceptibility pattern in the NICU of the Uberlândia University Hospital. METHODS: Data were collected through the National Healthcare Safety Network surveillance from January 2006 to December 2009. The patients were followed five times/week from their birth to their discharge or death. RESULTS: The study included 1,443 patients, 209 of these developed NIs, totaling 293 NI episodes, principally bloodstream infections (203; 69.3%) and conjunctivitis (52; 17.7%). Device-associated infection rates were as follows: 17.3 primary bloodstream infections per 1,000 central line-days and 3.2 pneumonias per 1000 ventilator-days. The mortality rate in neonates with NI was 11.9%. Mechanical ventilation, total parenteral nutrition, orogastric tube, previous antibiotic therapy, use of CVC and birth weight of 751-1,000g appeared to be associated with a significantly higher risk of NI (p < 0.05). In multiple logistic regression analysis for NI, mechanical ventilation and the use of CVC were independent risk factors (p < 0.05). Coagulase- negative Staphylococcus (CoNS) (36.5%) and Staphylococcus aureus (23.6%) were the most common etiologic agents isolated from cultures. The incidences of oxacillin-resistant CoNS and S. aureus were 81.8% and 25.3%, respectively. CONCLUSIONS: Frequent surveillance was very important to evaluate the association of these well-known risk factors with NIs and causative organisms, assisting in drawing the attention of health care professionals to this potent cause of morbidity. INTRODUÇÃO: Relatar a incidência das infecções hospitalares, microrganismos causadores, fatores de risco associados e o padrão de susceptibilidade aos antimicrobianos na UTI neonatal do Hospital Universitário de Uberlândia. MÉTODOS: Realizou-se vigilância National Healthcare Safety Network de janeiro de 2006 a dezembro de 2009. Os pacientes foram acompanhados cinco vezes por semana desde o seu nascimento até a alta ou óbito. RESULTADOS: O estudo incluiu 1.443 pacientes dos quais 209 desenvolveram infecção hospitalar, totalizando 293 episódios de IHs, com destaque para as infecções de corrente sanguínea (203; 69,3%) e conjuntivite (52; 17,7%). As taxas de infecção associadas a dispositivos foram as seguintes: 17,3 infecções da corrente sanguínea primária por 1.000 CVC dia e 3,2 pneumonias por 1.000 ventiladores-dia. A taxa de mortalidade em recém-nascidos com infecções hospitalares foi de 11,9%. A ventilação mecânica, nutrição parenteral total, sonda orogástrica, antibioticoterapia prévia, uso de CVC e peso de 751-1.000g pareceu estar associado com um risco significativamente maior de IH (p < 0,05). Na análise de regressão logística múltipla para IH, ventilação mecânica e uso de CVC foram fatores de risco independentes (p < 0,05). Staphylococcus coagulase-negativo (SCoN) (36,5%) e Staphylococcus aureus (23,6%) foram os agentes etiológicos mais comumente isolados. A incidência de SCoN e S. aureus resistente à oxacilina foram de 81,8% e 25,3%, respectivamente. CONCLUSÕES: Uma vigilância frequente foi importante para avaliar a associação de fatores de risco bem conhecidos com as IHs e etiologia com a finalidade de chamar a atenção dos profissionais de saúde para esta grande causa de morbidade.

On the basis of transmission electron microscopy observations in tumor cell lines, oncologists have made innumerous diagnostic and therapeutical progresses. Following this path, the UNICAMP immunopathologies laboratory established the NG97 cell line derived from a human astrocytoma grade III, which when injected to the athymic nude mouse flank developed a grade IV astrocytoma. In this study, we focused on ultrastructural characterization of the NG97 cells after being recovered from xenotransplant (NG97ht). These cells in culture were assayed by two different electron microscopy procedures to characterize ultrastructures related to grade IV astrocytomas and to observe their structures through cell subcultivation. Additionally, compara- tive morphological descriptions of different cell passages in these technical procedures could be a useful tool for improving electron microscopy cell lineage protocols. Results from many cell pas- sage observations showed ultrastructural similarities, which suggest malignant and glioblastoma phenotypes. In the first procedure, NG97ht cells were harvested and then incorporated into aga- rose before subjecting them to electron microscopy protocols, whereas in the second one, mono- layer cells grew first on cover slides. Comparison among protocols revealed that organelles, cyto- plasmatic extensions, spatial conformation of filopodia, and cell attachment to substrate were more preserved in the second procedure. Furthermore, in this latter procedure, a unique ellipsoi- dal structure was observed, which was already described when dealing with gliosarcoma cell line elsewhere. Therefore, these analyses demonstrated a morphological characterization of a new NG97ht cell line using electron transmission microscopy. Moreover, it has been shown that the second procedure provides more detailed information compared with the first. Microsc. Res. Tech.

Background The purpose of this study was evaluating the microbial flora of nurses' healthy (n = 30) and damaged hand (n = 30) by frequent handwashing and/or wearing of gloves. Methods Hand cultures were obtained both before and after washing hands with nonantimicrobial soap, through the sterile polyethylene bag method. Results The bacteria counts of the hands of professionals with damaged hands were higher than those with healthy hands, and those with damaged hands presented higher frequency of Staphylococcus aureus , 16.7% versus 10%; gram-negative bacteria, 20% versus 6.7%; and yeast, 26.7% versus 20%, respectively, as well as the sum of these microorganisms. The presence of Staphylococcus haemolyticus was only seen in nurses with damaged hands ( P = .02), and enterococci were not recovered from the hands of any volunteer. The presence of antimicrobial-resistant S aureus and gram-negative bacteria was also greater among damaged hands. Conclusion The irritation caused on the skin by frequent washing and/or wearing of gloves is associated with changes in hands microbial flora, and their potential risks should be considered when institutions/users are selecting products/formulations to assure hands skin health and consequent compliance with their own hygiene procedures.

The objective of this study was to evaluate clinical characteristics, etiology, and resistance to antimicrobial agents, among patients with ventilator-associated pneumonia (VAP). A case study vs. patients control under mechanical ventilation and hospitalized into clinical-surgical adults ICU of HC-UFU was performed from March/2005 to March/2006. Patients under ventilation for over 48 h were included in the study including 84 with diagnosis of VAP, and 191 without VAP (control group). Laboratory diagnosis was carried out through quantitative microbiological evaluation of tracheal aspirate. The identification of pathogens was performed by classical microbiological tests, and the antibiotics sensitivity spectrum was determined through the CLSI technique. VAP incidence rate over 1,000 days of ventilation was 24.59. The mean (+/- SD) duration of mechanical ventilation prior to VAP diagnosis was 23.2 +/- 17.2 days. By multivariate analysis the risk factors predisposing for VAP were: mechanical ventilation time and mechanical ventilation > seven days, tracheostomy and use of > or = three antibiotics. Mortality rate was high (32.1 %) but lower than that of the control group (46.5%). Major pathogens were identified in most of patients (95.2%) and included: Pseudomonas aeruginosa (29%), Staphylococcus aureus (26%), Enterobacter/Klebsiella/Serratia (19%) and Acinetobacter spp. (18%), with expressive frequencies of P. aeruginosa (52%), S. aureus (65.4%) and Enterobacteriaceae (43.7%) resistant to imipenem, oxacillin and 3/4 generation cephalosporins, respectively. In conclusion, our observation showed VAPs caused by multiresistant microorganisms, the prescription of > or = three antibiotics, and mortality with unacceptably high rates. The practice of de-escalation therapy appears to be urgently needed in order to improve the situation.

The emergence of Acinetobacter baumannii and Klebsiella pneumoniae strains in the hospital environment has been associated with the presence of multiple genetic elements, virulence factors and the ability to form biofilms. This study evaluated the biofilm formation ability of clinical and environmental A. baumannii and K. pneumoniae strains, isolated from various sources and presenting different molecular characteristics, resistance profiles and pulsed-field gel electrophoresis patterns. Fifty-three isolates were recovered from 2009 to 2014 in a Brazilian university hospital. Investigation of biofilm formation was performed for 10 strains of each species assessed by an initial adhesion assay, biofilm cell concentration and biofilm biomass, evaluated by quantitative assays in replicates, in three independent experiments. All strains of A. baumannii were able to attach to polystyrene plates, although two strains adhered to a lesser degree than the control. K. pneumoniae strains showed opposite behaviour, where only three strains adhered significantly when compared to the control. Quantitative evaluation revealed that in five A. baumannii and four K. pneumoniae isolates the biomass production could be characterised as moderate. None of the isolates were strong biofilm producers. Our results demonstrate: (1) biofilm formation is a heterogeneous property amongst A. baumannii and K. pneumoniae clinical strains and it was not associated with certain clonal types; (2) no relationship between multidrug resistance and biofilm production was observed; (3) more virulent K. pneumoniae strains tended to present higher production of biofilm.

This study evaluated the predictors of mortality and the impact of inappropriate therapy on the outcomes of patients with bacteraemia and ventilator-associated pneumonia (VAP). Additionally, we evaluated the correlation of the type III secretion system (TTSS) effector genotype with resistance to carbapenems and fluoroquinolones, mutations in the quinolone resistance-determining regions (QRDRs), metallo-β-lactamase and virulence factors. A retrospective cohort was conducted at a tertiary hospital in patients with multidrug-resistant (MDR) P. aeruginosa bacteraemia (157 patients) and VAP (60 patients). The genes for bla IMP, bla VIM, bla SIM, bla GIM and bla SPM and virulence genes (exoT, exoS, exoY, exoU, lasB, algD and toxA) were detected; sequencing was conducted for QRDR genes on fluoroquinolone-resistant strains. The multivariate analyses showed that the predictors independently associated with death in patients with bacteraemia were cancer and inappropriate therapy. Carbapenem resistance was more frequent among strains causing VAP (53.3 %), and in blood we observed the bla SPM genotype (66.6 %) and bla VIM genotype (33.3 %). The exoS gene was found in all isolates, whilst the frequency was low for exoU (9.4 %). Substitution of threonine to isoleucine at position 83 in gyrA was the most frequent mutation among fluoroquinolone-resistant strains. Our study showed a mutation at position 91 in the parC gene (Glu91Lys) associated with a mutation in gyrA (Thre83Ile) in a strain of extensively drug-resistant P. aeruginosa, with the exoT + exoS + exoU + genotype, that has not yet been described in Brazil to the best of our knowledge. This comprehensive analysis of resistance mechanisms to carbapenem and fluoroquinolones and their association with TTSS virulence genes, covering MDR P. aeruginosa in Brazil, is the largest reported to date.

We studied an outbreak of two multi-drug resistant clones of Acinetobacter baumannii in the Neonatal Intensive Care Unit of the Uberlândia Federal University Hospital in Minas Gerais state, Brazil, and we analyzed the contribution of cross-transmission in the rise in infection rates. Eleven neonates who developed multi-drug resistant A. baumannii nosocomial infection were matched to 22 neonates who were admitted to the same unit and did not develop an infection during the outbreak period, in order to identify risk factors for infection. Three out of the 11 neonates died. Epidemiological investigation included molecular typing, using pulsed field gel electrophoresis. Prior to the outbreak, from December 2001 to March 2002, no case of infection by this microorganism was diagnosed. Environmental and healthcare worker hand cultures were negative. Nine isolates had similar pulsed field gel electrophoresis patterns and two had another clone. The first clone was brought into the unit by an infected patient who was transferred from another hospital without a history of antibiotic use. The second clone did have its origin clearly defined. Both infected groups led us to conclude that several factors contributed to infection with A. baumannii. These factors were: exposure to antibiotics and invasive devices, birth weight1500 g, ageor = 7 days and duration of hospitalizationor = 7 days. Based on logistic regression, infected neonates were more exposed to carbapenem and mechanical ventilation than the control group. Cross transmission between infants contributed to the rise in the rates of multi-drug resistant A. baumannii infection.

Pulsed-field gel electrophoresis typing of 60 extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBLKp) isolates obtained in a neonatal intensive care unit during an outbreak indicated the dissemination of two major bacterial genotypes associated with colonization and invasive disease: one composed by aminoglycoside-resistant isolates and the other by aminoglycoside-susceptible isolates. A urease-negative phenotype was observed among aminoglycoside-resistant ESBLKp. Six pairs of isolates from gastrointestinal (GI) colonization and isolates from invasive disease that occurred 3-23 days later were shown to belong to the same genotype, reinforcing a direct association between colonization and subsequent disease. These data indicate that screening for ESBLKp GI colonization in an outbreak setting may be useful to detect neonates at a higher risk of invasive disease.

Seven of 50 Enterobacter cloacae strains from clinical isolates produced small turbid zones of hemolysis in horse and sheep blood agar plates, and the culture supernatants were also positive for hemolytic activity. The hem o l y sin was partially purified from the culture supernatant of E. cloacae by ultrafiltration (PM-10 membrane) and extraction with acetone. Semipurified hemolysin was stable to heating (100 °C, 30 min) and was soluble in organic solvents (acetone, ethanol, and methanol). The toxin showed no loss of biological activity after treatment with trypsin and was stable to acid treatment at pH 2.0 but not at a pH greater than 7.0. In the rat intestinal loop assay, the hemolysin caused hemorrhagic fluid accumulation and severe histological alterations. These findings indicate that this hemolysin may be a putative virulence factor in E. cloacae infections.Key words: Enterobacter cloacae, hemolysin, enterotoxin.

Bacterial counts were made of catheter insertion site and of catheter tips to help determine risk factors associated with catheterization of the jugular and subclavian veins. Among the 116 patients included in this study, 69% had central venous catheters (CVC) in the subclavian vein. Seven or more days catheterization (p=0.001) and > or =3 invasive devices (p=0.01) were infection risk factors associated with catheterization of the jugular vein. More than half of the patients presented high colony counts at the insertion site (> or =200 CFU/20 cm2) and 27% of the catheter tips were contaminated. The risk factors associated with contaminated catheter tips were > or =14 days hospital stay (p=0.02), > or =7 days catheterization (p=0.01) and antibiotic therapy (p=0.04). Coagulase-negative staphylococci (CoNS) and Staphylococcus aureus were the most common microorganisms at the insertion site (78%) and in the catheter tip (94%). Five patients presented sepsis (4.1%), four caused by Staphylococci and one by GNB. Twelve patients had the same microorganisms at the insertion site and catheter tip. We found a high prevalence of ORSA (62.5%) and ORCoNS (57.1%) in catheter tips. The high counts of staphylococci, including ORSA and ORCoNS, at the insertion site, and the significant association of this colonization with catheter tip contamination, indicate that the skin is an important reservoir of microorganisms associated with catheter-related bloodstream infection (CR-BSI). Health professionals should be aware of this potential source of infection at the CVC insertion site.

A cell-associated mannose-resistant hemagglutinating factor (HAF) was extracted from enteroinvasive Escherichia coli (EIEC) serotype O124:H- by sonication. Ultrastructural analysis of EIEC and immunocytochemical assays with the cell-free HAF and EIEC bacterial cells on HeLa cells, suggested that the HAF is a non-fimbrial putative adhesive factor that mediates in vivo adherence of EIEC to human epithelial cells.

The bacterial factors associated with bacteremia by multidrug-resistant and extensively drug-resistant P. aeruginosa, including overexpression of efflux pumps, AmpC overproduction, and loss/alteration of the OprD porin in isolates that are non-Metallo-β-Lactamase producing were analyzed in a retrospective study. Molecular analyses included strain typing by Pulsed Field Gel Electrophoresis and identification of key genes via qualitative and quantitative PCR-based assays. Previous use of carbapenems and tracheostomy was independently associated with the development of bacteremia by extensively drug-resistant and multidrug-resistant strains of P. aeruginosa. A high consumption of antimicrobials was observed, and 75.0% of the isolates contained amplicons with the blaSPM-1 and blaVIM genes, with a cross-dissemination of SPM-1. Of the 47 non-Metallo-β-Lactamase isolates, none had another type of carbapenemase. However, the isolates exhibited high rates of hyperproduction of AmpC, loss of the OprD porin (71.4%) and the presence of MexABOprM (57.1%) and MexXY (64.3%). This study suggests that in non-Metallo-β-Lactamase isolates, the association of intrinsic resistance mechanisms could contributes to the expression of multidrug-resistant/extensively drug-resistant phenotypes.

Objectives:To systematically review the literature on the occurrence of psychiatric diagnoses in a tinnitus-affected population, and correlate the presence of psychiatric disorders with tinnitus-related annoyance and severity.Method:A systematic review of the literature published between January 2000 and December 2012 was performed using PubMed, ISI Web of Science and SciELO databases. Original articles in English and Portuguese that focused on the diagnosis of mental disorders associated with tinnitus, especially anxiety and depression, were identified.Results:A total of 153 articles were found and 16 were selected. Fifteen articles showed a high prevalence of psychiatric disorders in tinnitus-affected patients, and nine showed a high correlation between the presence of a psychiatric disorder and tinnitus-related annoyance and severity.Conclusion:The prevalence of psychiatric disorders, especially anxiety and depression, is high in tinnitus patients, and the presence of these disorders correlates with tinnitus-related annoyance and severity.

The cytotoxic enterotoxin produced by Aeromonas hydrophila is considered to be the main virulence factor in gastrointestinal infections mediated by this pathogen. In this study, we examined the morphological and apoptotic effects of this toxin on HT29 cells, using light and electron microscopy in situ, as well as agarose gel electrophoresis of cell DNA. Cells treated with the cytotoxic enterotoxin became round and lost their polarity as well as their adhesion to each other and to the substrate. Cytoplasmic blebbing and nuclear condensation also occurred. DNA fragmentation was detected by TUNEL labelling and agarose gel electrophoresis. These results show that the cytotoxic enterotoxin of A. hydrophila can induce apoptosis in human intestinal cells in culture.Key words: Aeromonas hydrophila, cytotoxic enterotoxin, aerolysin, apoptosis, human intestinal cells.

We report a case-control study involving 663 primary orthopedic surgeries with a 12.7% rate of surgical site infections (SSIs). The respective frequencies of superficial incisional, deep incisional, and organ/space infections were 22.2%, 44.5%, and 33.3% for total hip arthroplasties; 66.7%, 33.3%, and 0% for total knee arthroplasties; 0%, 75.0% and 25.0% for hemiarthroplasties; and 44.1%, 35.3%, and 20.6% for osteosyntheses. These rates of SSIs are higher than those reported in previous studies.

INTRODUCTION : Antimicrobial resistance is an increasing threat in hospitalized patients, and inappropriate empirical antimicrobial therapy is known to adversely affect outcomes in ventilator-associated pneumonia (VAP). The aim of this study was to evaluate antimicrobial usage, incidence, etiology, and antimicrobial resistance trends for prominent nosocomial pathogens causing ventilator-associated pneumonia in a clinical-surgical intensive care unit (ICU). METHODS : Gram-negative bacilli and Staphylococcus aureus causing VAP, as well as their antimicrobial resistance patterns and data on consumption (defined daily dose [DDD] per 1,000 patient days) of glycopeptides, extended-spectrum cephalosporins, and carbapenems in the unit were evaluated in two different periods (A and B). RESULTS: Antimicrobial use was high, mainly of broad-spectrum cephalosporins, with a significant increase in the consumption of glycopeptides (p < 0.0001) and carbapenems (p < 0.007) in period B. For Acinetobacter baumannii and members of the Enterobacteriaceae family, 5.27- and 3.06-fold increases in VAPs, respectively, were noted, and a significant increase in resistance rates was found for imipenem-resistant A. baumannii (p = 0.003) and third-generation cephalosporins-resistant Enterobacteriaceae (p = 0.01) isolates in this same period. CONCLUSIONS: Our results suggest that there is a link between antibiotics usage at institutional levels and resistant bacteria. The use of carbapenems was related to the high rate of resistance in A. baumannii and therefore a high consumption of imipenem/meropenem could play a major role in selective pressure exerted by antibiotics in A. baumannii strains.

OBJECTIVE: to compare the degree of bacterial contamination of surgical instrument tables used in clean surgical procedures, either protected with plastic fields, sterilized with ethylene oxide, or disinfected with 70% alcohol and 1% iodine solutions. This is a randomized clinical trial in which samples were collected from the surfaces of surgical instrument tables before and after each procedure. Microbiological analysis was performed to identify microorganisms and their respective antimicrobial resistance. RESULTS: Bacterial growth in the surgeries using sterilized plastic was 5.71% before and 28.6% after surgery and, 2.9% and 45.7% respectively in surgeries using disinfection with 70% alcohol and 1% iodine solutions; no statistical difference was found between the methods. CONCLUSION: both methods present similar protection, however, 70% alcohol and 1% iodine do not generate solid waste. El objetivo del estudio fue analizar el grado de contaminación bacteriana de la mesa de instrumentos quirúrgicos en dos casos, después del uso: cubierta protectora plástico de la mesa quirúrgica esterilizada con óxido de etileno o desinfección con solución de alcohol a 70% y yodo a 1%, en procedimientos quirúrgicos limpios. Se trata de un experimento clínico aleatorio, con recolección de muestras de las superficies de las mesas de instrumentos quirúrgicos, antes y después de cada procedimiento, con posterior análisis microbiológica para identificar los microorganismos y la resistencia antimicrobiana. En las cirugías en que el plástico esterilizado fue utilizado, el crecimiento bacteriano fue de 5,71% antes y 28,6% después de la cirugía, en cuanto que en las desinfecciones con solución de alcohol a 70% y yodo a 1%, el crecimiento fue de 2,9% antes y 45,7% después, lo que indica que no hubo diferencia significativa entre los métodos empleados. Los dos métodos tienen poder de protección semejante, considerando que el alcohol a 70% y yodo a 1% no generan residuos sólidos. OBJETIVO: analisar o grau de contaminação bacteriana da mesa de instrumentais cirúrgicos, após o uso de campo plástico esterilizado por óxido de etileno ou a desinfecção com solução de álcool a 70% e iodo a 1%, em procedimentos cirúrgicos limpos. MÉTODOS: Trata-se de experimento clínico randomizado, com coletas de amostras das superfícies das mesas de instrumentais cirúrgicos, antes e depois de cada procedimento, com posterior análise microbiológica para identificação dos microrganismos e sua resistência antimicrobiana. RESULTADOS: nas cirurgias em que o plástico esterilizado foi utilizado, o crescimento bacteriano foi de 5,71% antes e 28,6% após a cirurgia, enquanto que nas desinfecções com solução de álcool a 70% e iodo a 1%, o crescimento foi de 2,9% antes e 45,7% após, sem diferença significativa entre os métodos empregados. CONCLUSÕES: os dois métodos têm poder de proteção semelhante, considerando que o álcool a 70% e iodo a 1% não geram resíduos sólidos.

To investigate the clonal diversity of Staphylococcus aureus strains isolated at João Pessoa, State of Paraíba, Brazil, digested genomic DNA were studied by pulsed-field gel electrophoresis (PFGE) in nine methicillin-resistant strains (MRSA) and three methicillin-sensitive strains (MSSA), selected among 67 isolates based on their antimicrobial susceptibility and epidemiology. The isolates were obtained between April and November 1992 from the Hospital of the Federal University of Paraíba, located in João Pessoa. Two MRSA isolates from the Oswaldo Cruz Hospital, São Paulo, Brazil, including an epidemic strain previously detected from different hospitals at the country were used as control. Five different patterns, were demonstrated by MRSA isolated in João Pessoa and these patterns were described in several epidemiologically unrelated hospitals in São Paulo. Our results suggest the interstate dissemination of a MRSA clone in João Pessoa which is similar to that described in other cities of Brazil.

The emergence of KPC-2 producing K. pneumoniae in hospitalized patients at the intensive care unit (ICU) of a teaching hospital located in the city of João Pessoa, Paraíba, Brazil, is reported. Seven carbapenem-resistant K. pneumoniae recovered from different body sites of infection were analyzed. Most isolates showed a multidrug-resistance phenotype. Genotypic analysis demonstrated the presence of two genotypes, with the predominance of genotype A, which belongs to ST 437. These isolates also carry the encoding genes of five other betalactamases. Universidade Federal de São Paulo (UNIFESP) Laboratório Alerta, Division of Infectious Diseases Universidade Federal da Paraíba Hospital Universitário Lauro Wanderley Universidade Federal de Uberlândia Biological Sciences Institute UNIFESP, Laboratório Alerta, Division of Infectious Diseases SciELO

Dact gene family encodes multifunctional proteins that are important modulators of Wnt and TGF-β signaling pathways. Given that these pathways coordinate multiple steps of limb development, we investigated the expression pattern of the two chicken Dact genes (Dact1 and Dact2) from early limb bud up to stages when several tissues are differentiating.During early limb development (HH24-HH30) Dact1 and Dact2 were mainly expressed in the cartilaginous rudiments of the appendicular skeleton and perichondrium, presenting expression profiles related, but distinct. At later stages of development (HH31-HH35), the main sites of Dact1 and Dact2 expression were the developing synovial joints. In this context, Dact1 expression was shown to co-localize with regions enriched in the nuclear β-catenin protein, such as developing joint capsule and interzone. In contrast, Dact2 expression was restricted to the interzone surrounding the domains of bmpR-1b expression, a TGF-β receptor with crucial roles during digit morphogenesis. Additional sites of Dact expression were the developing tendons and digit blastemas.Our data indicate that Dact genes are good candidates to modulate and, possibly, integrate Wnt and TGF-β signaling during limb development, bringing new and interesting perspectives about the roles of Dact molecules in limb birth defects and human diseases.