Expression of the locus of enterocyte effacement genes during the invasion process of the atypical enteropathogenic Escherichia coli 1711-4 strain of serotype O51:H40.

Atypical enteropathogenic Escherichia coli (aEPEC) is a significant cause of diarrhea in low- and middle-income countries. Certain aEPEC strains, including the Brazilian representative strain of serotype O51:H40 called aEPEC 1711-4, can use flagella to attach to, invade, and persist in T84 and Caco-2 intestinal cells. It can also translocate from the gut to extraintestinal sites in a rat model. Although various aspects of the virulence of this strain were studied and the requirement of a type III secretion system for the efficiency of the invasion process was demonstrated, the expression of the locus of enterocyte effacement (LEE) genes during the invasion and intracellular persistence remains unclear. To address this question, the expression of flagella and the different LEE operons was evaluated during kinetic experiments of the interaction of aEPEC 1711-4 with enterocytes in vitro. The genome of the strain was also sequenced. The results showed that flagella expression remained unchanged, but the expression of eae and escJ increased during the early interaction and invasion of aEPEC 1711-4 into Caco-2 cells, and there was no change 24 h post-infection during the persistence period. The number of actin accumulation foci formed on HeLa cells also increased during the 6-h analysis. No known gene related to the invasion process was identified in the genome of aEPEC 1711-4, which was shown to belong to the global EPEC lineage 10. These findings suggest that the LEE components and the intimate adherence promoted by intimin are necessary for the invasion and persistence of aEPEC 1711-4, but the detailed mechanism needs further study.IMPORTANCEAtypical enteropathogenic Escherichia coli (aEPEC) is a major cause of diarrhea, especially in low- and middle-income countries, like Brazil. However, due to the genome heterogeneity of each clonal group, it is difficult to comprehend the pathogenicity of this strain fully. Among aEPEC strains, 1711-4 can invade eukaryotic cells in vitro, cross the gut barrier, and reach extraintestinal sites in animal models. By studying how different known aEPEC virulence factors are expressed during the invasion process, we can gain insight into the commonalities of this phenotype among other aEPEC strains. This will help in developing preventive measures to control infections caused by invasive strains. No known virulence-encoding genes linked to the invasion process were found. Nevertheless, additional studies are still necessary to evaluate the role of other factors in this phenotype.

Estimation and probabilistic projection of age- and sex-specific mortality rates across Brazilian municipalities between 2010 and 2030.

BACKGROUND: Mortality rate estimation in small areas can be difficult due the low number of events/exposure (i.e. stochastic error). If the death records are not completed, it adds a systematic uncertainty on the mortality estimates. Previous studies in Brazil have combined demographic and statistical methods to partially overcome these issues. We estimated age- and sex-specific mortality rates for all 5,565 Brazilian municipalities in 2010 and forecasted probabilistic mortality rates and life expectancy between 2010 and 2030. METHODS: We used a combination of the Tool for Projecting Age-Specific Rates Using Linear Splines (TOPALS), Bayesian Model, Spatial Smoothing Model and an ad-hoc procedure to estimate age- and sex-specific mortality rates for all Brazilian municipalities for 2010. Then we adapted the Lee-Carter model to forecast mortality rates by age and sex in all municipalities between 2010 and 2030. RESULTS: The adjusted sex- and age-specific mortality rates for all Brazilian municipalities in 2010 reveal a distinct regional pattern, showcasing a decrease in life expectancy in less socioeconomically developed municipalities when compared to estimates without adjustments. The forecasted mortality rates indicate varying regional improvements, leading to a convergence in life expectancy at birth among small areas in Brazil. Consequently, a reduction in the variability of age at death across Brazil's municipalities was observed, with a persistent sex differential. CONCLUSION: Mortality rates at a small-area level were successfully estimated and forecasted, with associated uncertainty estimates also generated for future life tables. Our approach could be applied across countries with data quality issues to improve public policy planning.

Expression of the locus of enterocyte effacement genes during the invasion process of the atypical enteropathogenic Escherichia coli 1711-4 strain of serotype O51:H40.

Atypical enteropathogenic Escherichia coli (aEPEC) is a significant cause of diarrhea in developing countries. Some aEPEC strains, including the Brazilian representative strain of serotype O51:H40 called aEPEC 1711-4, can use flagella to attach to, invade, and persist in T84 and Caco-2 intestinal cells. They can even translocate from the gut to extraintestinal sites in a rat model. Although various aspects of the virulence of this strain were studied and the requirement of the T3SS for the efficiency of the invasion process was demonstrated, the expression of the LEE genes during the invasion and intracellular persistence remains unclear. To address this, the expression of flagella and the different LEE operons was evaluated during kinetic experiments of the interaction of aEPEC 1711-4 with enterocytes in vitro. The genome of the strain was also sequenced. The results showed that flagella expression remained unchanged, but the expression of eae and escJ increased during the early interaction and invasion of aEPEC 1711-4 into Caco-2 cells, and there was no change 24 hours post-infection during the persistence period. The number of pedestal-like structures formed on HeLa cells also increased during the 24-hour analysis. No known gene related to the invasion process was identified in the genome of aEPEC 1711-4, which was shown to belong to the global EPEC lineage 10. These findings suggest that LEE components and the intimate adherence promoted by intimin are necessary for the invasion and persistence of aEPEC 1711-4, but the detailed mechanism needs further study.

A spatiotemporal analysis of bovine brucellosis cases in Minas Gerais state, Brazil, from 2011 to 2018.

Our study explored the patterns of bovine brucellosis dissemination in Minas Gerais state, Brazil, by examining data on passive surveillance of bovine brucellosis cases from the Instituto Mineiro de Agropecuaria (IMA) (Animal Health Authority), as well as cattle population and bovine brucellosis testing, from 2011 to 2018 by means of a spatiotemporal analysis. We plotted cases, populations and testing distributions and performed spatial autocorrelation (Moran's I test) and local indicators of spatial autocorrelation (LISA) analyses. Moreover, we assessed the correlation of the spatial distribution and the compiled data (brucellosis cases, cattle populations, and brucellosis testing) by Lee's test. Our results showed that bovine brucellosis cases occurred mainly in the Triângulo Mineiro, Alto Paranaíba and Northwest regions, which reported cases in all analyzed years (2011 to 2018). The cattle population of Minas Gerais was concentrated in the same regions as bovine brucellosis cases, and the performed tests through the analyzed years (2011 to 2018). Moran's I test results of the case data showed significant spatial autocorrelation in 2011, 2015 and 2018 (p value < 0.05), and from 2011 to 2018, the population and testing data were also significant in Moran's I test (p value < 0.01). The results of cluster analysis (LISA) of cases showed clusters mainly in the Triângulo Mineiro, Alto Paranaíba, Northwest and South regions in 2011, 2015 and 2018. The local clusters for cattle populations and brucellosis testing were also observed in the same regions as bovine brucellosis cases in all years (2011 to 2018). The correlation results between clusters (Lee's test) were 0.22 (p value < 0.01) in 2011, 0.15 (p value < 0.01) in 2015 and 0.43 (p value <0.01) in 2018 between cases and populations, and 0.25 (p value <0.01) in 2011, 0.14 (p value <0.01) in 2015 and 0.38 (p value < 0.01) in 2018 for testing and cases. Therefore, our results showed that brucellosis cases were distributed together with cattle populations and brucellosis testing data, indicating that brucellosis in cattle in Minas Gerais state is being identified where there are more animals and where more tests are performed.

Comparing modified Lee and White method against 20-minute whole blood clotting test as bedside coagulation screening test in snake envenomation victims.

Background: Twenty-minute whole blood clotting test (20WBCT) and Modified Lee and White (MLW) method are the most routinely employed bedside tests for detecting coagulopathic snake envenomation. Our study compared the diagnostic utility of MLW and 20WBCT for snakebite victims at a tertiary care hospital in Central Kerala, South India. Methods: This single-center study recruited 267 patients admitted with snake bites. 20WBCT and MLW were performed simultaneously at admission along with the measurement of Prothrombin Time (PT). The diagnostic utility of 20WBCT and MLW was determined by comparing the sensitivity (Sn), specificity (Sp), positive and negative predictive values, likelihood ratios, and accuracy at admission with an INR value > 1.4. Results: Out of 267 patients, 20 (7.5%) patients had VICC. Amongst those who had venom-induced consumption coagulopathy (VICC), MLW was prolonged for 17 patients, (Sn 85% 95% confidence interval [CI]: 61.1-96.0) whereas 20WBCT was abnormal for 11 patients (Sn 55%, 95% CI: 32.04-76.17). MLW and 20WBCT were falsely positive for the same patient (Sp 99.6%, 95% CI: 97.4-99.9%). Conclusion: MLW is more sensitive than 20WBCT to detect coagulopathy at the bedside amongst snakebite victims. However, further studies are necessary for standardizing bedside coagulation tests in snakebite cases.

GrlR, a negative regulator in enteropathogenic E. coli, also represses the expression of LEE virulence genes independently of its interaction with its cognate partner GrlA.

Introduction: Enteropathogenic Escherichia coli (EPEC), enterohemorrhagic E. coli (EHEC) and Citrobacter rodentium (CR) belong to a group of pathogens that share the ability to form "attaching and effacing" (A/E) lesions on the intestinal epithelia. A pathogenicity island known as the locus of enterocyte effacement (LEE) contains the genes required for A/E lesion formation. The specific regulation of LEE genes relies on three LEE-encoded regulators: Ler activates the expression of the LEE operons by antagonizing the silencing effect mediated by the global regulator H-NS, GrlA activates ler expression and GrlR represses the expression of the LEE by interacting with GrlA. However, despite the existing knowledge of LEE regulation, the interplay between GrlR and GrlA and their independent roles in gene regulation in A/E pathogens are still not fully understood. Methods: To further explore the role that GrlR and GrlA in the regulation of the LEE, we used different EPEC regulatory mutants and cat transcriptional fusions, and performed protein secretion and expression assays, western blotting and native polyacrylamide gel electrophoresis. Results and discussion: We showed that the transcriptional activity of LEE operons increased under LEE-repressing growth conditions in the absence of GrlR. Interestingly, GrlR overexpression exerted a strong repression effect over LEE genes in wild-type EPEC and, unexpectedly, even in the absence of H-NS, suggesting that GrlR plays an alternative repressor role. Moreover, GrlR repressed the expression of LEE promoters in a non-EPEC background. Experiments with single and double mutants showed that GrlR and H-NS negatively regulate the expression of LEE operons at two cooperative yet independent levels. In addition to the notion that GrlR acts as a repressor by inactivating GrlA through protein-protein interactions, here we showed that a DNA-binding defective GrlA mutant that still interacts with GrlR prevented GrlR-mediated repression, suggesting that GrlA has a dual role as a positive regulator by antagonizing GrlR's alternative repressor role. In line with the importance of the GrlR-GrlA complex in modulating LEE gene expression, we showed that GrlR and GrlA are expressed and interact under both inducing and repressing conditions. Further studies will be required to determine whether the GrlR alternative repressor function depends on its interaction with DNA, RNA, or another protein. These findings provide insight into an alternative regulatory pathway that GrlR employs to function as a negative regulator of LEE genes.

Comparing modified Lee and White method against 20-minute whole blood clotting test as bedside coagulation screening test in snake envenomation victims

Abstract Background: Twenty-minute whole blood clotting test (20WBCT) and Modified Lee and White (MLW) method are the most routinely employed bedside tests for detecting coagulopathic snake envenomation. Our study compared the diagnostic utility of MLW and 20WBCT for snakebite victims at a tertiary care hospital in Central Kerala, South India. Methods: This single-center study recruited 267 patients admitted with snake bites. 20WBCT and MLW were performed simultaneously at admission along with the measurement of Prothrombin Time (PT). The diagnostic utility of 20WBCT and MLW was determined by comparing the sensitivity (Sn), specificity (Sp), positive and negative predictive values, likelihood ratios, and accuracy at admission with an INR value > 1.4. Results: Out of 267 patients, 20 (7.5%) patients had VICC. Amongst those who had venom-induced consumption coagulopathy (VICC), MLW was prolonged for 17 patients, (Sn 85% 95% confidence interval [CI]: 61.1-96.0) whereas 20WBCT was abnormal for 11 patients (Sn 55%, 95% CI: 32.04-76.17). MLW and 20WBCT were falsely positive for the same patient (Sp 99.6%, 95% CI: 97.4-99.9%). Conclusion: MLW is more sensitive than 20WBCT to detect coagulopathy at the bedside amongst snakebite victims. However, further studies are necessary for standardizing bedside coagulation tests in snakebite cases.

Whole-genome sequencing analysis of Shiga toxin-producing Escherichia coli O22:H8 isolated from cattle prediction pathogenesis and colonization factors and position in STEC universe phylogeny.

Shiga toxin-producing Escherichia coli (STEC) is a foodborne pathogen capable of causing illness in humans. In a previous study, our group showed that a STEC isolate belonging to O22:H8 serotype (strain 154) can interfere with STEC O157:H7 colonization both in vitro and in vivo. Using whole-genome sequencing and genomic comparative, we predicted a subset of genes acquired by O22:H8 strain 154 through horizontal gene transfer that might be responsible for the phenotype previously described by our group. Among them were identified genes related to the pathogenesis of non-LEE (locus of enterocyte effacement) STEC, specific metabolic processes, antibiotic resistance and genes encoding for the T6SS-1 that is related to inter-bacterial competition. In addition, we showed that this strain carries stx1c and stx2dact, a mucus-inducible variant. The results obtained in this study provide insights into STEC genomic plasticity and the importance of genomic islands in the adaptation and pathogenesis of this pathogen.

Biofilm formation by LEE-negative Shiga Toxin-Producing Escherichia coli strains.

Shiga toxin-producing Escherichia coli (STEC) include several serotypes isolated from cases of hemorrhagic colitis and, hemolytic uremic syndrome. Although O157:H7 is the most predominant STEC serotype, more than 100 non-O157 serogroups cause diseases in humans. Some STEC carry a Locus of Enterocyte Effacement (LEE-positive); however, STEC that do not carry LEE (LEE-negative) have also been associated with illness, mainly those harbouring the Locus of Adhesion and Autoaggregation (LAA). LAA carry some genes such as hes, iha, tpsA, and agn43, related with pathogenicity. One of them is the ability to form biofilms on different environments, which can contaminate food and generate infections while protecting themselves against adverse conditions. Considering that LAA could be responsible for some adherence mechanisms, the aims of this study were to compare different serogroup of LEE-negative STEC strains in their ability to form biofilms and to evaluate the participation of some genes encoding in LAA. A total of 348 LEE-negative STEC strains was analyzed. The presence of hes, iha, tpsA and agn43 were determined by monoplex PCR. From them, 48 STEC strains belonging to serogroups O113, O130, O171, O174 and, O178 were assayed for their ability to form biofilm. The most prevalent genes detected were agn43 (72.1%) and tpsA (69.5%). The iha and hes genes were present in 63.7% and 54% of the strains, respectively. Although all STEC strains were able to form biofilm, it was found a high variability between them. The relation between the biofilm formation and the presence of each gene was not statistically significant, suggesting that biofilm formation is independent of the presence of those genes. Highlighting that there is no treatment for HUS, it is once again notable that prevention measures and control strategies to prevent biofilm formation are important factors in reducing STEC transmission.

Differential expression of the virulence gene nleB among Shiga toxin-producing Escherichia coli strains.

Shiga toxin-producing Escherichia coli (STEC) is a zoonotic foodborne pathogen associated with hemolytic uremic syndrome (HUS) that vary in their ability to cause disease in humans. STEC represents a serious problem for public health and Argentina is the country with the highest HUS incidence worldwide. Non-LEE effector (nle) genes, present on pathogenicity islands (PAIs), encode translocated substrates of the type III secretion system (T3SS), which could have an important role in STEC virulence. Particularly, nleB is one of the main effector genes proposed as a virulence marker that is involved in the action of T3SS during the STEC infection. NleB inhibits the inflammatory response of the host cell allowing the bacteria to persist in the first stage of the infection. In order to identify the potential risk of STEC strains for public health, the aim of this study was to evaluate and compare basal nleB transcription of 24 STEC strains belonging to 10 serotypes isolated from cattle, food and patients. The results showed differences in nleB transcription among strains. Some non-O157:H7 strains presented transcription levels above the control, an O157:H7 HUS-producing strain. On the other hand, no significant differences were found in basal transcription levels associated with origin or serotype but differences were found between HUS and non-HUS strains. These differences in nleB transcription may be of importance in STEC pathogenesis and could help to differentiate high and low virulence STEC strains.

ELF Magnetic Field Exposure System for In Vitro Studies Based on Lee-Whiting Coils.

In order to run a series of in vitro studies on the effect of extremely low-frequency magnetic fields on cell cultures, developing and characterizing an appropriate exposure system is required. The present design is based on a two-shielded Lee-Whiting coils system. The circular design was chosen because its axial symmetry allowed for both reducing simulation unknowns and measurement points during the characterization, and additionally made the machining of the parts easier. The system can generate magnetic flux densities (B fields) up to 1 mT root-mean-square amplitude (rms) with no active cooling system in the incubator, and up to 3 mTrms with it. The double-wrapped windings with twisted pairs allow for the use of each set of coils either as exposure or control with no detectable parasitic B field in the control. The artifacts have also been analyzed; the B field in the center of the sham control chamber is about 1 µTrms for a maximum of 3 mTrms in the exposure chamber, the parasitic incident electric fields are less than 1 V/m, the temperature difference between sham and exposure chamber is less than or equal to 0.2 °C, and the typical vibration difference between sham and exposure is less than 0.1 m/s2 . © 2020 Bioelectromagnetics Society.

Atuação fonoaudiológica na doença de Machado-Joseph: relato de caso / Effect of speech-language therapy in Machado-Joseph disease: case report

RESUMO A doença de Machado-Joseph é a forma de ataxia espinocerebelar de maior prevalência no Brasil e tem como alguns dos principais sinais clínicos a disfagia e a disartria. Este relato de caso objetivou verificar os efeitos da intervenção intensiva fonoaudiológica em um paciente com a doença de Machado-Joseph. A coleta de dados foi realizada a partir de protocolos de avaliação de fala e deglutição e protocolos de autoavaliação de qualidade de vida, em relação à deglutição e comunicação. Também foram realizadas avaliações quantitativas de parâmetros acústicos. A intervenção foi administrada por meio do método Lee Silverman, programa intensivo que visa ao aumento da intensidade vocal. A partir das avaliações clínicas e instrumentais, os resultados demonstraram melhora em todas as bases motoras de fala, respiratória, fonatória, ressonantal, articulatória e a prosódia, além da diminuição dos sinais disfágicos. Na qualidade vocal, houve diminuição de rouquidão e instabilidade, regularização de jitter e shimmer, aumento da intensidade vocal, melhora na coordenação de palavras e frases por expiração e, ainda, melhora discreta da diadococinesia. Após intervenção, a autoavaliação de qualidade de vida relacionada à deglutição apresentou valores iguais ou maiores nos domínios diretamente ligados à alimentação, porém, os domínios emocionais diminuíram. O paciente relatou satisfação em todos os domínios da qualidade de vida em voz e foram obtidos valores maiores em todos os domínios. Concluiu-se que a intervenção intensiva beneficiou o participante e impactou positivamente sua qualidade de vida.

ABSTRACT Machado-Joseph disease is the most prevalent form of spinocerebellar ataxia in Brazil, and has dysphagia and dysarthria among its main clinical signs. This case report aims to ascertain the effects of intensive speech-language intervention in a patient with Machado-Joseph disease. Data collection was performed based on speech and swallowing assessment protocols and self-assessment protocols specific to swallowing-related and communication-related quality of life. Quantitative assessments of acoustic parameters were also performed. The intervention was administered through the Lee Silverman method, which is an intensive program aimed at increasing vocal intensity. The results of clinical and instrumental evaluations showed improvement in all motor parameters of speech (respiration, phonation, resonance, articulation, and prosody), besides a reduction in dysphagic signs. Regarding vocal quality, there was a decrease in hoarseness and instability, regularization of jitter and shimmer, increased vocal intensity, and improved coordination of words and phrases by expiration, as well as slight improvement of diadochokinesis. After intervention, self-assessment of swallowing-related quality of life was unchanged or improved in the domains directly related to food, but reduced in emotional domains. The patient reported satisfaction in all domains of voice-related quality of life, and scores were increased in all domains. We conclude that intensive intervention was beneficial for the participant and positively impacted their quality of life.

Cumulative acquisition of pathogenicity islands has shaped virulence potential and contributed to the emergence of LEE-negative Shiga toxin-producing Escherichia coli strains.

Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens causing severe gastroenteritis, which may lead to hemolytic uremic syndrome. The Locus of Enterocyte Effacement (LEE), a Pathogenicity Island (PAI), is a major determinant of intestinal epithelium attachment of a group of STEC strains; however, the virulence repertoire of STEC strains lacking LEE, has not been fully characterized. The incidence of LEE-negative STEC strains has increased in several countries, highlighting the relevance of their study. In order to gain insights into the basis for the emergence of LEE-negative STEC strains, we performed a large-scale genomic analysis of 367 strains isolated worldwide from humans, animals, food and the environment. We identified uncharacterized genomic islands, including two PAIs and one Integrative Conjugative Element. Additionally, the Locus of Adhesion and Autoaggregation (LAA) was the most prevalent PAI among LEE-negative strains and we found that it contributes to colonization of the mice intestine. Our comprehensive and rigorous comparative genomic and phylogenetic analyses suggest that the accumulative acquisition of PAIs has played an important, but currently unappreciated role, in the evolution of virulence in these strains. This study provides new knowledge on the pathogenicity of LEE-negative STEC strains and identifies molecular markers for their epidemiological surveillance.

Mortality forecasting in Colombia from abridged life tables by sex.

BACKGROUND: An adequate forecasting model of mortality that allows an analysis of different population changes is a topic of interest for countries in demographic transition. Phenomena such as the reduction of mortality, ageing, and the increase in life expectancy are extremely useful in the planning of public policies that seek to promote the economic and social development of countries. To our knowledge, this paper is one of the first to evaluate the performance of mortality forecasting models applied to abridged life tables. OBJECTIVE: Select a mortality model that best describes and forecasts the characteristics of mortality in Colombia when only abridged life tables are available. DATA AND METHOD: We used Colombian abridged life tables for the period 1973-2005 with data from the Latin American Human Mortality Database. Different mortality models to deal with modeling and forecasting probability of death are presented in this study. For the comparison of mortality models, two criteria were analyzed: graphical residuals analysis and the hold-out method to evaluate the predictive performance of the models, applying different goodness of fit measures. RESULTS: Only three models did not have convergence problems: Lee-Carter (LC), Lee-Carter with two terms (LC2), and Age-Period-Cohort (APC) models. All models fit better for women, the improvement of LC2 on LC is mostly for central ages for men, and the APC model's fit is worse than the other two. The analysis of the standardized deviance residuals allows us to deduce that the models that reasonably fit the Colombian mortality data are LC and LC2. The major residuals correspond to children's ages and later ages for both sexes. CONCLUSION: The LC and LC2 models present better goodness of fit, identifying the principal characteristics of mortality for Colombia.Mortality forecasting from abridged life tables by sex has clear added value for studying differences between developing countries and convergence/divergence of demographic changes.

First report of the distribution of Locus of Adhesion and Autoaggregation (LAA) pathogenicity island in LEE-negative Shiga toxin-producing Escherichia coli isolates from Argentina.

Shiga toxin-producing Escherichia coli (STEC) are important foodborne pathogens that can cause severe disease. The ability to adhere to epithelial cells is an important virulence trait and pathogenicity islands (PAIs) play an important role. Recently, researchers identified a member of the Heat-resistant agglutinin family and characterized this antigen named Hemagglutinin from Shiga toxin-producing E. coli (Hes). More importantly, they showed that hes and other genes such as iha, pagC and agn43 were integrated in each of the four modules present in the new PAI named Locus of Adhesion and Autoaggregation (LAA) whose presence is associated with severe disease linked to with LEE-negatives STEC. The distribution of LAA among STEC strains isolates from different origins between 2000 and 2015 from cattle, the farm environment, and food and harboring diverse virulence was investigated. The STEC strains were characterized by PCR to detect three modules of LAA and agn43 (as marker of module IV), and phylogenetic groups were determined. LAA was found in 46% of LEE-negative STEC corresponding to serogroups O91, O174, O113, O171, O178, O130 and others. The presence of this PAI is associated with strains harboring stx2 (56%) and belonging to phylogroup B1 (91%). LAA is a novel pathogenicity island associated with strains isolated from Hemolytic Uremic Syndrome cases. Therefore, the results of this study contribute to a better understanding regarding the pathogenicity of this emergent subset of STEC strains harboring LAA as a predictor of virulence of LEE-negative STEC strains.

Translation, Cross-Cultural Adaptation, and Validation of the Lee Chronic Graft-versus-Host Disease Symptom Scale in a Brazilian Population.

The Lee Chronic Graft-versus-Host Disease (GVHD) Symptom Scale is a patient-reported instrument developed and validated in English to measure the symptoms and functional impact of cGVHD. This tool has not yet been validated in a Latin American population, however. The Brazil-Seattle Chronic GVHD Consortium conducted a multicenter study at 5 Brazilian institutions to validate the Lee cGVHD Symptom Scale in adults with cGVHD. Study objectives included the translation and validation of the instrument in Brazilian Portuguese and evaluation of the correlation with other quality of life (QoL) tools, including the Medical Outcomes Study Short Form 36 (SF-36) and Functional Assessment of Chronic Illness Therapy with Bone Marrow Transplant subscale (FACT-BMT). Translation and validation were done according to the American Association of Orthopedic Surgeons Outcome Committee guidelines. Spearman's correlation coefficient was used to measure construct validity. Reliability was assessed using Cronbach's α and intraclass correlation coefficients. Between April 2011 and August 2012, 47 patients with cGVHD based on the 2005 National Institutes of Health criteria (29 males [62%], 18 females [38%]; median age, 48 years; range, 23 to 69 years) were enrolled in this study. The reliability of the Lee cGVHD Symptom Scale was adequate (Cronbach's α = 0.62 to 0.83). The correlations between similar domains of the Lee cGVHD Symptom Scale, SF-36, and FACT-BMT were moderate to high. Our data indicate that the Brazilian Portuguese version of the Lee cGVHD Symptom Scale is valid and reliable and can be used in clinical trials of cGVHD in Brazil.

Vaccines for viral and bacterial pathogens causing acute gastroenteritis: Part II: Vaccines for Shigella, Salmonella, enterotoxigenic E. coli (ETEC) enterohemorragic E. coli (EHEC) and Campylobacter jejuni.

In Part II we discuss the following bacterial pathogens: Shigella, Salmonella (non-typhoidal), diarrheogenic E. coli (enterotoxigenic and enterohemorragic) and Campylobacter jejuni. In contrast to the enteric viruses and Vibrio cholerae discussed in Part I of this series, for the bacterial pathogens described here there is only one licensed vaccine, developed primarily for Vibrio cholerae and which provides moderate protection against enterotoxigenic E. coli (ETEC) (Dukoral(®)), as well as a few additional candidates in advanced stages of development for ETEC and one candidate for Shigella spp. Numerous vaccine candidates in earlier stages of development are discussed.

Sensibilidad, especificidad y valores predictivos de los índices cardíacos de Goldman, Detsky y Lee / Sensitivity, specificity and predictive values of the Goldman, Detsky and Lee cardiac indices

Introducción: Reconocer oportunamente las variables de riesgo perioperatorio permite predecir la frecuencia de morbimortalidad, así como tomar medidas a fin de reducir complicaciones. Para ello se han creado varias escalas de riesgo en pacientes portadores de enfermedad cardiovascular. Objetivo: Determinar la sensibilidad, la especificidad y los valores predictivos de los índices de riesgo cardíaco de Goldman, Detsky y Lee para cirugía no cardíaca. Método: Se realizó un estudio observacional, analítico, longitudinal y prospectivo del total de pacientes portadores de enfermedad cardiovascular con enfermedad quirúrgica no cardíaca en el período comprendido entre enero de 2011 y enero de 2013 en el Hospital Universitario Manuel Ascunce Domenech de la ciudad de Camagüey. La muestra estuvo constituida por 88 pacientes comprendidos en el universo que cumplieron con los criterios de inclusión. Las variables estudiadas fueron: edad, sexo, tipo de cirugía, tipo de complicación, y la presencia o no de estas en relación con el riesgo catalogado según los índices de Goldman, Detsky y Lee. Se aplicó prueba de sensibilidad, especificidad y valores predictivos. Resultados: Predominaron los pacientes mayores de 70 años, el sexo masculino, la cirugía ortopédica; la arritmia cardíaca fue la principal complicación. Fue frecuente encontrar pacientes de alto riesgo, en los cuales la mayoría sufrieron complicaciones. Conclusiones: Los índices de Goldman y Detsky mostraron alta sensibilidad y especificidad, y el índice de Lee, mayor valor predictivo positivo. No obstante, deben aplicarse los 3 índices predictivos para lograr una óptima estratificación del riesgo cardíaco en cirugía no cardíaca.

Introduction: Timely recognition of perioperative risk variables helps predict morbidity and mortality frequency, as well as adopt measures to reduce complications. Several risk scores have been developed for this purpose in patients with cardiovascular disease. Objective: To determine the sensitivity, specificity and predictive values of the Goldman, Detsky and Lee cardiac risk indices for non-cardiac surgery. Methods: Observational, analytical, longitudinal prospective study of the total number of patients with cardiovascular disease undergoing non-cardiac surgery between January 2011 and January 2013 at Hospital Universitario Manuel Ascunce Domenech in Camagüey. The sample consisted of 88 patients included in the universe of patients who met the inclusion criteria. The variables studied were: age, gender, type of surgery, type of complication, and the presence or absence of complications in relation to the risk assessed on the basis of the Goldman, Detsky and Lee indices. The sensitivity, specificity and predictive value test was applied. Results: There was a predominance of males in patients over 70 years of age coming for orthopaedic surgery; cardiac arrhythmiawas the main complication. High-risk patients were a frequent finding and the majority suffered complications. Conclusions: The Goldman and Detsky indices showed high sensitivity and specificity, while the Lee index showed higher positive predictive value. However, the three predictive indices must be applied in order to optimize cardiac risk stratification in non-cardiac surgery.

Avaliação da interação de Escherichia coli enterohemorrágica (EHEC) pertencente ao sorotipo O157: H7 isoladas de bovinos assintomáticos e de doença humana com células enterocíticas humanas (linhagem Caco-2) / Evaluation of interaction of Enterohaemorrhagic (EHEC) 0157: H7 isolated from asymptomatic cattle and human disease with human enterocitic cells

Reconhecida como agente de doença humana em 1982, E.coli enterohemorrágica (EHEC) pode causar diarréia sanguinolenta, colite hemorrágica e síndrome hemolítica urêmica (SHU). EHEC constitui um subgrupo especialmente virulento das E.coli produtoras de toxina de Shiga (Stx). O fator crítico da sua virulência é a toxina Shiga, capaz de interromper a síntese proteica da célula eucariótica. São conhecidos dois subgrupos de Stx, Stx1 e Stx2. Stx1 possui duas variantes Stx1c e Stx1d. Stx2 possui muitas variantes. Estudos epidemiológicos sugerem que cepas com os perfis toxigênicos Stx2 ou Stx2/Stx2c seriam mais frequentemente associadas a pacientes com SHU. Além da expressão de Stx, EHEC do sorotipo O157:H7 colonizam a mucosa intestinal induzindo a formação de lesões denominadas ‘attaching/effacing’ (A/E). Para a produção da lesão A/E, é necessária a presença de uma ilha de patogenicidade cromossômica denominada LEE, composta por cinco operons, LEE 1 a LEE5. Em LEE 5 são codificadas a adesina intimina e o seu receptor Tir, o qual é translocado por um sistema de secreção tipo III (SSTT) e em LEE 4 são codificadas as proteínas secretadas EspA,B e D. Em EHEC O157:H7 são descritos muitos fatores de virulência, codificados em ilhas de patogenicidade, no cromossomo e no megaplasmídio pO157. Bovinos são o principal reservatório deste patógeno e alimentos de origem bovina e produtos contaminados com fezes de bovinos são causadores de surtos epidêmicos. Em nosso país EHEC O157:H7 é isolada do reservatório animal mas é muito rara a sua ocorrência em doença humana. Notamos que nas cepas bovinas predomina Stx2c, enquanto nas cepas humanas predomina o perfil toxigenico Stx2/Stx2c...

Recognized in 1982 as a human pathogen, enterohemorrhagic Escherichia coli (EHEC) causes bloody diarrhea, hemorrhagic colitis and hemolytic uremic syndrome (HUS). EHEC belonging to serotype O157:H7 are mostly important in North America, United Kingdom and Japan. Shiga toxin (Stx) is the critical factor of STEC. Stx is capable to interrupt the protein synthesis of the eukaryotic cell. Two subgroups of Stx are known, Stx1 and Stx2. Two variants of Stx1 are known (Stx1c and Stx1d), but several Stx2 variants have been described. Epidemiological studies suggest that STEC/EHEC strains carrying the toxigenic profiles Stx2 or Stx2/Stx2c are more frequently associated to HUS. Besides the expression of Stx, EHEC O157:H7 colonize the intestinal mucosa inducing the formation of characteristic histopathological lesions denominated “attaching/effacing” (A/E). To the production of A/E lesions, it is necessary the presence of a pathogenicity island called LEE (locus of enterocyte effacement), composed by five operons, LEE 1 to LEE5. An outer membrane adhesin (intimin) and its receptor Tir, which is translocated by a type three secretion sytem (TTSS), are both codified in LEE5 while the secreted proteins EspA, B and D, that constitute part of the SSTT, are codified in LEE4. Cattle are the main reservoir of this pathogen and foods of bovine origin and products contamined with bovine feces are common causes of epidemic outbreaks. In Brazil, EHEC O157:H7 can be isolated from the animal reservoir . Stx2c prevails among the bovine strains, while the toxigenic profiles Stx2 or Stx2/Stx2c are found among the human strains...