Protocolo clínico de atendimento de pneumonia adquirida na comunidade para Pronto-Socorro Infantil do Hospital do Servidor Público Municipal de São Paulo / Clinical protocol for community-acquired pneumonia care for the Children's Emergency Room of the Hospital do Servidor Público Municipal de São Paulo

A pneumonia adquirida na comunidade (PAC) é a infecção aguda do parênquima pulmonar que ocorre no meio comunitário. A PAC representa a maior causa de morbidade e mortalidade em todo o mundo em crianças abaixo de cinco anos. Nesta faixa etária, a etiologia viral é a mais comum; porém, dentre as causas bacterianas, o Streptoccocus pneumoniae é o mais prevalente. As manifestações clínicas variam de acordo com o patógeno, hospedeiro e da gravidade da doença, sendo geralmente descrita com tosse, febre e desconforto respiratório. A PAC complicada é a pneumonia que, apesar do uso de antibióticos, evolui com complicações locais ou sistêmicas. Nos pacientes hospitalizados, as hemoculturas devem ser consideradas para auxiliar no diagnóstico etiológico e planejamento terapêutico. O tratamento inicial deve ser iniciado empiricamente com antibióticos. Caso haja necessidade de hospitalização, hemoculturas devem ser consideradas para auxiliar na propedêutica. Após implementação das vacinas pneumocócicas, principalmente após introdução da vacina pneumocócica 13 valente (PCV 13), houve redução significativa dos casos de pneumonia bacteriana e também da necessidade hospitalização. Diante de tal realidade, a elaboração do trabalho possui como objetivo a melhora dos procedimentos e a padronização dos atendimentos da população pediátrica com um quadro clínico sugestivo pneumonia adquirida na comunidade, que procura o serviço de Pronto Atendimento Infantil do Hospital do Servidor Público Municipal de São Paulo (HSPM), ao construir um protocolo clínico de atendimento específico para a doença. O presente trabalho objetiva elaborar um protocolo clínico de atendimento de pneumonia adquirida na comunidade no Hospital do Servidor Público Municipal de São Paulo, contribuindo na assistência médica dos pacientes pediátricos. Apesar do grande avanço com a introdução das vacinas pneumocócicas, a PAC ainda representa uma importante causa de mortalidade na população infantil, sendo fundamental a elaboração de protocolos clínicos para abordar corretamente os pacientes que recorrem a um Pronto Socorro Infantil. Protocolos clínicos são diretrizes fundamentadas nas melhores práticas para a abordagem e tratamento de determinadas doenças, baseadas em evidência científica. O presente trabalho objetiva a melhora dos procedimentos e a uniformização dos atendimentos da população pediátrica com pneumonia, que procura o serviço de Pronto Atendimento Infantil do Hospital do Servidor Público Municipal de São Paulo (HSPM), com a construção de um protocolo clínico de atendimento específico para a doença, a partir da revisão de literatura atualizada, cujo período de vigência seguirá os progressos científicos sobre o tema. Palavras-chave: Pneumonia Adquirida da Comunidade. Protocolos clínicos. Pediatria. Serviços Médicos de Emergência. Vacinas Pneumocócicas

Human Milk Concentrations of Minerals, Essential and Toxic Trace Elements and Association with Selective Medical, Social, Demographic and Environmental Factors.

This study aims to quantify concentrations of minerals and trace elements in human milk (HM) and infant formula (IF) and evaluate associations with medical, social, environmental, and demographic variables. A prospective, case series study of 170 nursing mothers was made. HM samples were obtained from full-term (colostrum, intermediate and mature HM) and preterm (mature HM) mothers. Variables of interest were assessed by a questionnaire. For comparison, IF samples (n = 30) were analyzed in a cross-sectional study. Concentrations of 35 minerals, essential and toxic trace elements were quantified, 5 for the first time: thallium in HM and IF; strontium in preterm HM; and gallium, lithium and uranium in IF. In preterm and full-term HM, levels of selenium (p < 0.001) were significantly lower than recommended and were associated with low birth weight (p < 0.002). Cesium and strontium concentrations were significantly higher than recommended (p < 0.001). Associations were observed between arsenic and residence in an urban area (p = 0.013), and between lead and smoking (p = 0.024) and well-water consumption (p = 0.046). In IF, aluminum, vanadium, and uranium levels were higher than in HM (p < 0.001); uranium, quantified for the first time, was 100 times higher in all types of IF than in HM. Our results indicate that concentrations of most trace elements were within internationally accepted ranges for HM and IF. However, preterm infants are at increased risk of nutritional deficiencies and toxicity. IF manufacturers should reduce the content of toxic trace elements.

Avaliação da atividade antimicrobiana e citotoxicidade hemolítica em diferentes extratos vegetais / Evaluation of antimicrobial activity and hemolytic cytotoxicity in different plant extracts

Introdução: A busca por novos agentes terapêuticos tem incentivado as pesquisas com plantas medicinais, pois muitas delas podem apresentar propriedade antimicrobiana e conhecer o potencial citotóxico dos extratos é fundamental para garantir a segurança durante o uso. Objetivo: Avaliar a atividade antimicrobiana e a citotoxicidade hemolítica de Arctium lappa (bardana), Equisetum arvense (cavalinha), Mikania glomerata (guaco), Morus nigra (amora) e Plantago major (tanchagem), amplamente consumidos pela população na forma de chás medicinais. Material e Métodos: Os extratos etanólicos fora preparados a 20% por percolação. Na avaliação antimicrobiana foi utilizada a técnica de difusão em disco, empregando as bactérias Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Acinetobactersp, Enterococcus sp e Salmonella sp. O ensaio de citotoxicidade baseou-se na exposição dos extratos a 5%, 25%, 50%, 75% e 100% em suspensão de hemácias a 37ºC por 30 minutos, seguido de centrifugação e visualização do grau de hemólise. Resultados: Todos os extratos apresentaram inibição de crescimento microbiano, principalmente sobre Acinetobacter sp (amora), Enterococcus sp (amora e cavalinha), K. pneumoniae(amora, bardana e guaco), P. aeruginosa (cavalinha, tanchagem, bardana e guaco) e Salmonellasp (amora e bardana). No ensaio de citotoxicidade, o grau de hemólise foi classificado como baixo para tanchagem e bardana (5%) e médio para cavalinha, guaco e amora (25%) nas concentrações testadas. Conclusão: Os resultados mostram o potencial antimicrobiano dos extratos de amora, bardana, cavalinha, guaco e tanchagem contra bactérias Gram negativas e a baixa citotoxicidade hemolítica confirma a segurança no uso dos mesmos como agentes terapêuticos.

Infectious Diseases in Free-Ranging Blonde Capuchins, Sapajus flavius, in Brazil

The main threats to primates worldwide are the degradation, fragmentation,and loss of their habitats; hunting (especially for bushmeat); and illegal trade.For many species, the most important threat is forest fragmentation, resulting in small populations that are restricted to isolated forest patches. In this situation, primates are particularly vulnerable to disease. The Endangered blonde capuchin (Sapajus flavius) is now restricted to a few forest patches in Northeast Brazil. We investigated the occurrence of parasites and bacterial diseases in one of three free-ranging groups of S. flavius in a small forest patch in Paraíba state, Northeast Brazil. We tested for antibodies against Leishmania spp., Trypanosoma cruzi, Toxoplasma gondii, Leptospira spp. (24 strains), and Brucella spp.. We used molecular analysis to detect Plasmodium spp., and evaluated blood smears for the presence of hemoparasites. All individuals tested negative for Leptospira spp. and B. abortus, but 8 of 48 (16%) presented antibodies for both Leishmania spp. and T. cruzi. We identified antibodies to T. gondii in 12% of the individuals tested. Plasmodium brasilianum infection was present in 4% of the individuals tested, and blood smears showed microfilariae parasites in 46% of the individuals tested...

Prevalencia de bacterias enteropatógenas en niños que asisten a un hospital pediátrico en Resistencia, Chaco, Argentina / Prevalence of enteropathogenic bacteria in children attending a pediatric hospital in Resistencia, Chaco, Argentina

La enfermedad diarreica aguda continúa siendo uno de los problemas de salud pública más serios en los países en desarrollo, en los que constituye una de las causas principales de enfermedad y muerte en los niños menores de 5 años. Su epidemiología es totalmente dependiente de la región geográfica, nivel socio económico, costumbres y hábitos de la población.El objetivo del presente trabajo fue determinar la prevalencia de agentes etiológicos bacterianos causantes de diarrea aguda, en niños atendidos en un Hospital Pediátrico de Resistencia, Chaco, en el año 2013.Se investigó la presencia de Shigella spp., Salmonella spp., Campylobacte rspp., Escherichia coli O157:H7 en muestras de materia fecal de niños con enfermedad diarreica aguda. Sobre 823 muestras de materia fecal analizadas en el período mencionado, 93 resultaron positivas para alguno de los enteropatógenos estudiados (Tasa de recuperación del 11,3%).Las frecuencias de aislamiento de los enteropatógenos fueron: Shigella spp (82,8%), Salmonella spp (9,7%), Campylobacter spp (6,5%), y E. coli O157:H7 (1%).Con respecto a las especies, dentro del género Shigella predominó S. flexneri (60/77) seguida de S. sonnei (13/77) y S. boydii (4/77). Con excepción de E. coli O157, en el presente trabajo no se estudiaron los diferentes tipos patogénicos.Como en el resto del país, S. flexneri continúa siendo el agente etiológico más frecuentemente aislado. Este es el primer informe sobre la presencia de Campylobacter en coprocultivos en la provincia del Chaco.

Acute diarrheal disease remains one of the most serious problems of public health in developing countries, which is one of the leading causes of illness and death in children under 5 years. Its epidemiology is totally dependent on the geographic region, socioeconomic status, customs and habits of the population.The aim of this study was to determine the prevalence of bacterial etiologic agents causing acute diarrhea in children attending a Pediatric Hospital in the city of Resistencia, Chaco, during 2013.In this work Shigella spp, Salmonella sp, Campylobacter spp, Escherichia coli O157:H7 were investigatedAmong 823 stool samples analyzed, 93 were positive for any of the enteropathogens studied (recovery rate 11.3%).The frequency of isolation of enteric pathogens were: Shigella spp (82.8%), Salmonella spp (9.7%), Campylobacter spp (6.5%), and E. coli O157: H7 (1%).Respect to genus Shigella, Shigella flexneri was the prevalent (60/77) followed by S. sonnei (13/77) and S. boydii (4/77). With the exception of E. coli O157 in the present work the other pathogenic types were not studied.As in the rest of the country, S. flexneri remains the most frequently isolated etiologic agent. This is the first report about the presence of Campylobacter in stool cultures in the province of Chaco

Study on the kinetic characteristics of trace harmful gases for a two-person-30-day integrated CELSS test.

A two-person-30-day controlled ecological life support system (CELSS) integrated test was carried out, and more than 30 kinds of trace harmful gases including formaldehyde, benzene, and ammonia were measured and analyzed dynamically. The results showed that the kinds and quantities of the trace harmful gases presented a continuously fluctuating state during the experimental period, but none of them exceed the spacecraft maximum allowable concentration (SMAC). The results of the Pre-Test (with two persons without plants for 3 days) and the Test (with two persons and four kinds of plants for 30 days) showed that there are some notable differences for the compositions of the trace harmful gases; the volatile organic compounds (VOCs) such as toluene, hexane, and acetamide were searched out in the Pre-Test, but were not found in the Test. Moreover, the concentrations of the trace harmful gases such as acetic benzene, formaldehyde, and ammonia decreased greatly in the Test more than those in the Pre-Test, which means that the plants can purify these gases efficiently. In addition, the VOCs such as carbon monoxide, cyclopentane, and dichloroethylene were checked out in the Test but none in the Pre-Test, which indicates that these materials might be from the crew's metabolites or those devices in the platform. Additionally, the ethylene released specially by plants accumulated in the later period and its concentration reached nearly ten times of 0.05 mg m(-3) (maximum allowed concentration for plant growth, which must have promoted the later withering of plants). We hoped that the work can play a referring function for controlling VOCs effectively so that future more CELSS integrating tests can be implemented smoothly with more crew, longer period, and higher closure.

Levels of selected carcinogens and toxicants in vapour from electronic cigarettes.

SIGNIFICANCE: Electronic cigarettes, also known as e-cigarettes, are devices designed to imitate regular cigarettes and deliver nicotine via inhalation without combusting tobacco. They are purported to deliver nicotine without other toxicants and to be a safer alternative to regular cigarettes. However, little toxicity testing has been performed to evaluate the chemical nature of vapour generated from e-cigarettes. The aim of this study was to screen e-cigarette vapours for content of four groups of potentially toxic and carcinogenic compounds: carbonyls, volatile organic compounds, nitrosamines and heavy metals. MATERIALS AND METHODS: Vapours were generated from 12 brands of e-cigarettes and the reference product, the medicinal nicotine inhaler, in controlled conditions using a modified smoking machine. The selected toxic compounds were extracted from vapours into a solid or liquid phase and analysed with chromatographic and spectroscopy methods. RESULTS: We found that the e-cigarette vapours contained some toxic substances. The levels of the toxicants were 9-450 times lower than in cigarette smoke and were, in many cases, comparable with trace amounts found in the reference product. CONCLUSIONS: Our findings are consistent with the idea that substituting tobacco cigarettes with e-cigarettes may substantially reduce exposure to selected tobacco-specific toxicants. E-cigarettes as a harm reduction strategy among smokers unwilling to quit, warrants further study. (To view this abstract in Polish and German, please see the supplementary files online.).

Potencial probiótico in vitro de bactérias ácido-láticas isoladas de queijo-de-minas artesanal da Serra da Canastra, MG / Probiotic potential of lactic acid bacteria isolated from minas artisanal cheese from Serra da Canastra, MG

O potencial probiótico in vitro de 12 amostras de bactérias ácido-láticas (11 Lactobacillus spp. e uma Weissella paramesenteroides), isoladas de queijo-de-minas artesanal da Serra da Canastra, foi investigado. Essas amostras foram caracterizadas in vitro quanto à susceptibilidade a antimicrobianos, antagonismo contra microrganismos de referência patogênicos e não patogênicos e sensibilidade a pH gástrico e sais biliares. Apenas L. rhamnosus B4, W. paramesenteroides C10 e L. rhamnosus D1 apresentaram resistência a menor número de antimicrobianos (ceftazidima, oxacilina, estreptomicina e vancomicina). Todas as amostras apresentaram atividade antagonista frente a todas as bactérias patogênicas testadas e não inibiram as bactérias não patogênicas. Todas as amostras foram resistentes ao pH gástrico (2.0). Alguns microrganismos mostraram pouca inibição do crescimento em presença de sais biliares, enquanto outros foram moderadamente ou altamente inibidos. L. rhamnosus B4, W. paramesenteroides C10 e L.rhamnosus D1 apresentaram o melhor potencial probiótico de acordo com os testes in vitro realizados.

The in vitro probiotic potential of twelve samples of lactic acid bacteria (eleven Lactobacillus spp. and one Weissella paramesenteroides) isolated from Minas artisanal cheese from Serra da Canastra was evaluated for their antimicrobial susceptibility, antagonistic activity against pathogenic and lactic acid bacteria, as well as for their tolerance to gastric pH and biliary salts. Only L. rhamnosus B4, W. paramesenteroides C10, and L. rhamnosus D1 showed resistance to tested antimicrobials (ceftazidime, oxacillin, streptomycin, and vancomycin). All samples showed antagonism against all pathogenic bacteria tested and did not inhibit the growth of nonpathogenic samples. Regarding gastric pH tolerance, all samples showed low inhibition at pH 2. The tolerance to biliary salts varied from low to great among tested samples. L. rhamnosus B4, W. paramesenteroides C10, and L. rhamnosus D1 showed the best probiotic potential revealed by their in vitro test results.

Biowarfare and bioterrorism.

Bioterrorism is not only a reality of the times in which we live but bioweapons have been used for centuries. Critical care physicians play a major role in the recognition of and response to a bioterrorism attack. Critical care clinicians must be familiar with the diagnosis and management of the most likely bioterrorism agents, and also be adequately prepared to manage a mass casualty situation. This article reviews the epidemiology, diagnosis, and treatment of the most likely agents of biowarfare and bioterrorism.

A single-blinded, single-centre, controlled study in healthy adult smokers to identify the effects of a reduced toxicant prototype cigarette on biomarkers of exposure and of biological effect versus commercial cigarettes.

BACKGROUND: Despite universal acceptance that smoking is harmful, a substantial number of adults continue to smoke. The development of potential reduced exposure products (more recently termed modified risk tobacco products) has been suggested as a way to reduce the risks of tobacco smoking. This trial is designed to investigate whether changes in toxicant exposure after switching from a commercial to reduced toxicant prototype (RTP) cigarette (7 mg International Organisation for Standardisation (ISO) tar yield) can be assessed by measurement of biomarkers and other factors. The primary objective is to descriptively assess changes in selected biomarkers of exposure (BoE) and biomarkers of biological effect (BoBE) within participants and within and between groups after switching. Secondary objectives are to assess similarly changes in other biomarkers, quality of life, smoking behaviours, physiological measures, mouth-level exposure to toxicants and sensory perception. METHODS/DESIGN: This trial will assess current smokers, ex-smokers and never-smokers in a single-centre single-blind, controlled clinical trial with a forced-switching design and in-clinic (residential) and ambulatory (non-residential) periods. Smokers will be aged 23-55 years (minimum legal smoking age plus 5 years) and non-smokers 28-55 years (minimum legal smoking age plus 5 years, plus minimum 5 years since last smoked). Smokers will be allowed to smoke freely at all times. We will assess changes in selected BoE and BoBE and effective dose in urine and blood after switching. Creatinine concentrations in serum, creatinine clearance in urine, cotinine concentration in saliva, diaries and collection of spent cigarette filters will be used to assess compliance with the study protocol. Mouth-level exposure to toxins will be assessed by filter analysis. DISCUSSION: Data from this study are expected to improve scientific understanding of the effects of RTP cigarettes on BoE and BoBE, and give insights into study design for clinical assessment of potential MRTPs. TRIAL REGISTRATION: The study was registered in the Current Controlled Trials database under the reference ISRCTN81286286.

Methodologies for the quantitative estimation of toxicant dose to cigarette smokers using physical, chemical and bioanalytical data.

Methodologies have been developed, described and demonstrated that convert mouth exposure estimates of cigarette smoke constituents to dose by accounting for smoke spilled from the mouth prior to inhalation (mouth-spill (MS)) and the respiratory retention (RR) during the inhalation cycle. The methodologies are applicable to just about any chemical compound in cigarette smoke that can be measured analytically and can be used with ambulatory population studies. Conversion of exposure to dose improves the relevancy for risk assessment paradigms. Except for urinary nicotine plus metabolites, biomarkers generally do not provide quantitative exposure or dose estimates. In addition, many smoke constituents have no reliable biomarkers. We describe methods to estimate the RR of chemical compounds in smoke based on their vapor pressure (VP) and to estimate the MS for a given subject. Data from two clinical studies were used to demonstrate dose estimation for 13 compounds, of which only 3 have urinary biomarkers. Compounds with VP > 10(-5) Pa generally have RRs of 88% or greater, which do not vary appreciably with inhalation volume (IV). Compounds with VP < 10(-7) Pa generally have RRs dependent on IV and lung exposure time. For MS, mean subject values from both studies were slightly greater than 30%. For constituents with urinary biomarkers, correlations with the calculated dose were significantly improved over correlations with mouth exposure. Of toxicological importance is that the dose correlations provide an estimate of the metabolic conversion of a constituent to its respective biomarker.

Isolation and identification of main mastitis pathogens in Mexico / Isolação e identificação dos micróbios patogênicos principais da mastite em México

The present work is a large epidemiological study aiming to detect the prevalence of subclinical mastitis and to investigate the major udder pathogens in Jalisco State, western Mexico. For this purpose, 2205 dairy cows, representing 33 Mexican dairy herds, were involved. Of 2205 cows, 752 mastitic animals were diagnosed and only 2,979 milk samples could be obtained for further investigation. All 2979 milk samples were subjected to California Mastitis Test (CMT) to differentiate clinical cases from subclinical ones where 1996 samples (67 %) reacted positively. Of these, 1087 samples (54.5%) came from cows suffering from clinical cases of mastitis. Bacteriological identification of the causative agents revealed the presence of a major group of pathogens including the Coagulase negative staphylococci (CNS), S.aureus, S.agalactiae, Corynebacterium spp. and Coliform bacteria which were detected in 464 (15.6%), 175 (5.9%), 200 (6.8%), 417 (14%) and 123 (4.1%) of the 2927 investigated quarters, 295 (15.4%), 118 (15.7%), 111 (14.8%), 227 (30.2%) and 109 (14.5%) of the 752 examined cows and in 33 (100%), 22 (66.7%), 19 (57.6%), 30 (90.1%) and 27 (81.8%) of the 33 herds involved, respectively. Other pathogens could be detected in the investigated milk samples such as S. dysgalactiae (0.4%), S.uberis (0.37%), Bacillus spp. (1%), Nocardia spp. (0.6%) und Candida spp. (0.1%). Meanwhile, others were present in a negligible ratio; including the Aerococcus viridans, and Enterococcus spp., Lactococcus lactis, S. bovis.

O trabalho atual é um estudo epidemiológico que objetiva detectar a predominância da mastite subclínica e investigar os micróbios patogênicos principais do úbere no México ocidental. Com esta finalidade, foram utilizadas 2205 vacas leiteiras, representando 33 rebanhos de leiteiras mexicanas. Além dessas 2205 vacas, 752 animais com mastite foram diagnosticados, considerando-se que somente 2979 amostras do leite poderiam ser obtidas para a posterior investigação. Todas as 2979 amostras do leite foram submetidas ao teste da mastite de Califórnia (CMT) para diferenciar casos clínicos dos subclínicos, visto que 1996 amostras (67%) reagiram positivamente. Além dessas, 1087 amostras (54.5%) vieram das vacas que sofrem de casos clínicos de mastite. A identificação bacteriológica dos agentes causais revelou a presença dos Staphylococcus negativos para coagulase (CNS), S. aureus, S. agalactiae, outros spp. Streptococcal, Corynebacterium spp., e as bactérias de coliformes foram detectadas em 464 (15.6%), 175 (5.9%), 200 (6.8%), 109 (3.9%), 417 (14%) e em 123 (4.1%) dos 2927 quartos investigados; em 295 (15.4%), 118 (15.7%), 111 (14.8%), 95 (12.6%), 227 (30.2%) e em 109 (14.5%) das 752 vacas examinadas e, finalmente, em 33 (100%), 22 (66.7%), 19 (57.6%), 30 (90.1%), 30 (90.1%) e em 27 (81.8%) dos 33 rebanhos envolvidos, respectivamente.

Changes in levels of biomarkers of exposure observed in a controlled study of smokers switched from conventional to reduced toxicant prototype cigarettes.

UNLABELLED: Reduced toxicant prototype (RTP) cigarettes with substantially reduced levels of tobacco smoke toxicants have been developed. Evaluation of these prototype cigarettes included measurement of biomarkers of exposure (BoE) to toxicants in smokers switched from conventional cigarettes to the RTPs. A 6-week single-blinded randomised controlled study with occasional clinical confinement was conducted ( TRIAL REGISTRATION: ISRCTN7215735). All smoking subjects smoked a conventional cigarette for 2-weeks. Control groups continued to smoke the conventional cigarette while test groups switched to one of three RTP designs. Clinical confinement and additional assessments were performed for all smoking groups after 2 and 4-weeks. A non-smoker group provided background levels of BoE. On average, smokers switched to RTPs with reduced machine yields of toxicants had reduced levels of corresponding BoEs. For vapour phase toxicants such as acrolein and 1,3-butadiene reductions of ⩾70% were observed both in smoke chemistry and BoEs. Reductions in particulate phase toxicants such as tobacco-specific nitrosamines, aromatic amines and polyaromatic hydrocarbons depended upon the technologies used, but were in some cases ⩾80% although some increases in other particulate phase toxicants were observed. However, reductions in BoEs demonstrate that it is possible to produce prototype cigarettes that reduce exposure to toxicants in short-term use.

Molecular epidemiology of Listeria monocytogenes isolated from different sources in Brazil / Epidemiologia molecular de Listeria monocytogenes isoladas de diferentes fontes no Brasil

Listeria monocytogenes is an important foodborne pathogen that primarily affects pregnant women, neonates, the elderly and immune-compromised individuals, and it may cause abortion, septicemia, and meningitis. From the 13 capsular groups described, serotypes 4b, 1/2b and 1/2a are most closely related to human infection. For this reason, serotyping has limited value as an epidemiological tool; thus, improved discriminatory typing methods are required to enhance knowledge of L. monocytogenes contamination and infection. The aim of this study was to characterize the genetic diversity of L. monocytogenes isolates in the pork processing industry in Sao Paulo, Brazil and human infection isolates by ERICPCR and single enzyme AFLP. Serotypes 1/2c and 4b were frequent among isolates from pork and slaughterhouse/market environments, whereas serotypes 4b and 1/2a were observed among human isolates. ERIC-PCR and AFLP revealed 34 and 31 distinct profiles, respectively, which had tendencies of separation according to serogroup and isolate origin. The genetic profiles from slaughterhouse and market environments suggest the possibility of different sources of Listeria contamination in the environment, although in certain cases, continuous contamination caused by the persistence of clonal strains is also a possibility.

Listeria monocytogenes é um importante patógeno de origem alimentar que afeta principalmente grávidas, neonatos, idosos e indivíduos imunocomprometidos, e pode causar abortamento, septicemia e meningite. Dos 13 grupos capsulares descritos, os sorotipos 4b, 1/2b e 1/2a são os mais relacionados à infecção humana. Por esta razão, a sorotipagem possui valor limitado como ferramenta epidemiológica e, dessa forma, métodos mais discriminatórios são necessários para melhorar o conhecimento sobre a contaminação e a infecção por L. monocytogenes. O objetivo deste estudo foi caracterizar a diversidade genética de isolados de L. monocytogenes da indústria de processamento de carne suína noEstado de São Paulo, Brasil, e compará-los a isolados de casos de infecção humana através do ERIC-PCR e AFLP com uma única enzima. Os sorotipos 1/2c e 4b foram frequentes em carne suína e ambientes de abatedouros e mercados, enquanto os sorotipos 4b e 1/2a foram observados nos isolados de humanos. ERIC-PCR e AFLP resultaram em 34 e 31 perfis distintos, respectivamente, com uma tendência a separar de acordo com o sorogrupo e a origem do isolado. Os perfis genéticos de ambiente dos abatedouros e mercados sugerem a possibilidade de diferentes origens de contaminação por Listeria nos ambientes estudados, porém, em alguns casos, é possível que ocorra a persistência de cepas clonais causando contaminação contínua.

Development and evaluation of a novel subunit vaccine for Mycoplasma gallisepticum / Desenvolvimento e avaliação de uma nova vacina para Mycoplasma gallisepticum

Adhesion proteins from Mycoplasma gallisepticum (MG) encoded by cytadhesion genes mgc1 and mgc2 were cloned into plasmid vectors and transformed into E. coli. Seventeen groups of specific-pathogen free (SPF), birds at four weeks of age were used to inoculate these two proteins (MGC1 and MGC2) mixed into an oil emulsion creating a novel MG vaccine. Six different protein concentrations (50, 100, 200, 400, 800, and 1000µg/bird) were tested with two equal concentration doses at four and seven weeks of age. In addition, many control groups were needed such as bacterin, membrane, no vaccine or challenge, oil emulsion alone, and no vaccine but challenged. Three weeks following the second vaccination, 50% of the birds in each treatment group were challenged with MG strain S6. The remaining birds were left as contacts to verify protection against horizontal transmission. All birds were bled before vaccinations, challenge and euthanasia. Birds were negative for MG at the first vaccination, as shown by serum plate agglutination test. At necropsy, tissue samples (trachea, lungs, and air sacs) were collected for histopathological examination. Swabs from trachea were used for PCR analysis. ELISA results showed a strong immune response to both protein preparations and almost the same response level for different doses tested, proving the immunogenic features of MGC1 and MGC2. However, humoral responses failed to prevent MG infection and disease when challenged as demonstrated by PCR and histopathology. MGC1 contact birds showed some degree of infection by PCR analysis. In addition, histopathological and ELISA results suggest that contact birds did not have enough time to develop lesions and to mount an immune response.

Os genes mgc1 e mgc2, codificadores de duas proteínas de adesão (MGC1 e MGC2) da bactéria Mycoplasma gallisepticum, foram clonados em E. coli. Dezessete grupos de aves livres de patógenos específicos (SPF), com quatro semanas de idade, foram inoculados com uma emulsão oleosa contendo as proteínas MGC1 e MGC2 purificadas. Seis concentrações (50, 100, 200, 400, 800, e 1000µg/ave) foram testadas com duas doses idênticas, às quatro e sete semanas de vida, respectivamente. Além disso, grupos controles foram avaliados com uma vacina comercial contra micoplasmose aviária, membrana de MG, grupo sem vacina/sem desafio, grupo vacina oleosa de MGC1 sem desafio, grupo com vacina oleosa de MGC2 sem desafio, grupo desafiado mas sem vacina. Três semanas após a segunda e a última vacinação, 50% dos animais dos grupos tratamentos foram desafiados com a cepa S6 de MG. O restante dos animais foi deixado como contato para averiguar proteção contra a transmissão horizontal da doença. Amostras de sangue de todas as aves foram coletadas antes das vacinações, do desafio e da eutanásia. As aves eram negativas para MG às quatro semanas de vida, conforme visto na aglutinação em placa. Na necropsia, tecidos (traqueia, pulmão e sacos aéreos) foram coletados para exame histopatológico. Suabes da traqueia foram utilizados para a PCR. Os resultados do ELISA demonstraram forte resposta imune contra as duas proteínas testadas e resposta similar independentemente do número de doses, provando a sua capacidade imunogênica. Porém, esta resposta humoral gerada foi incapaz de prevenir a infecção e a doença após desafio, conforme demonstrado pelos exames PCR e histopatológico. Aves-contato, inoculadas com MGC1, demonstraram estar infectadas nas análises de PCR. Além disso, os resultados do histopatológico e ELISA sugerem que os animais-contato não tiveram tempo suficiente para demonstrar lesões ou resposta imune.

Adductomics: characterizing exposures to reactive electrophiles.

To understand environmental causes of disease, unbiased methods are needed to characterize the human exposome, which represents all toxicants to which people are exposed from both exogenous and endogenous sources. Because they directly modify DNA and important proteins, reactive electrophiles are probably the most important constituents of the exposome. Exposures to reactive electrophiles can be characterized by measuring adducts from reactions between circulating electrophiles and blood nucleophiles. We define an 'adductome' as the totality of such adducts with a given nucleophilic target. Because of their greater abundance and residence times in human blood, adducts of hemoglobin (Hb) and human serum albumin (HSA) are preferable to those of DNA and glutathione for characterizing adductomes. In fact, the nucleophilic hotspot represented by the only free sulfhydryl group in HSA (HSA-Cys(34)) offers particular advantages for adductomic experiments. Although targeted adducts of HSA-Cys(34) have been monitored for decades, an unbiased method has only recently been reported for visualizing the HSA-Cys(34) 'subadductome'. The method relies upon a novel mass spectrometry application, termed fixed-step selected reaction monitoring (FS-SRM), to profile Cys(34) adducts in tryptic digests of HSA. Here, we selectively review the literature regarding the potential of adductomics to partially elucidate the human exposome, with particular attention to the HSA-Cys(34) subadductome.

Atividade antimicrobiana de óleos essenciais em bactérias patogênicas de origem alimentar / Antimicrobial activity of essential oils against sessile and planktonic pathogens of food source

Objetivou-se identificar e quantificar os constituintes e avaliar a atividade antimicrobiana dos óleos essenciais de Mentha piperita, Cymbopogon citratus, Ocimum basilicum e Origanum majorana contra cepas de Escherichia coli enteropatogênica, Salmonella enterica Enteritidis, Listeria monocytogenes e Enterobacter sakazaki. A obtenção dos óleos essenciais foi realizada a partir de folhas secas, empregando-se a técnica de hidrodestilação e utilizando-se a aparelho de Clevenger modificado. A atividade antibacteriana dos óleos essenciais foi determinada pelo método de difusão em ágar. Observou-se que os óleos essenciais inibiram o crescimento bacteriano, mas a efetividade foi variada. Entre os óleos essenciais testados, M. piperita apresentou maior atividade antibacteriana para E. coli, (8.106 UA mL-1) quando comparada as demais bactérias, atividade moderada para Salmonella enterica Enteritidis e Enterobacter sakazakii (1.706 e 3.200 UA mL-1 respectivamente) e baixa atividade para Listeria monocytogenes (106,67 UA mL-1). Já óleo essencial de Cymbopogon citratus apresentou maior atividade antimicrobiana frente a E. coli (9.386 UA mL-1) e atividade moderada frente a Enterobacter sakazakii, Salmonella enterica Enteritidis e Listeria monocytogenes (2.773 UA mL-1 para ambas). Ocimum basilicum apresentou maior atividade antibacteriana frente E. coli e Enterobacter sakazakii (6.826 e 8.106 UA mL-1 respectivamente), moderada atividade frente a Salmonella enterica Enteritidis (1.600 UA mL-1) e não apresentou atividade frente a Listeria monocytogenes.Origanum majorana também foi testado neste estudo e apresentou maior atividade antimicrobiana frente E. coli (5.973 UA mL-1), atividade moderada para Salmonella enterica Enteritidis e Enterobacter sakazakii (1.706 e 2.346 UA mL-1 , respectivamente) e não apresentou atividade para Listeria monocytogenes.

ABSTRACT The objective of this work was to identify and quantify the constituents, and to evaluate the antimicrobial activity of the essential oils from Mentha piperita, Cymbopogon citratus, Ocimum basilicum and Origanum majorana, against enteropathogenic Escherichia coli, Salmonella enterica Enteritidis, Listeria monocytogenes and Enterobacter sakazakii. The essential oils were obtained from dried leaves by using the hydrodistillation technique and the modified Clevenger apparatus, and their bacterial activity was determined by using the agar diffusion technique. The essential oils inhibited bacterial growth, but their effectiveness was varied. Among the essential oils tested, that from M. piperita showed a greater antimicrobial activity against E. coli (8.106 UA mL-1), moderate activity for S. enterica Enteritidis and E. sakazakii (1.706 e 3.200 UA mL-1 respectively) and low activity for L. monocytogenes (106,67 UA mL-1). However, the essential oil from C. citratus presented a greater antimicrobial activity against E. coli (9.386 UA mL-1) and a moderate activity against E. sakazakii, S. enterica Enteritidis and L. monocytogenes (2.773 UA mL-1 for both). The essential oil from O. basilicum showed a greater antimicrobial activity against E. coli and E. sakazakii (6.826 e 8.106 UA mL-1 respectively),moderate activity against S. enterica Enteritidis (1.600 UA mL-1), and was inactive against L, monocytogenes. Origanum majorana, which was also tested in our work, showed a greater antibacterial activity against E. coli, (5.973 UA mL-1) moderate activity against S. enterica Enteritidis and E. sakazakii (1.706 e 2.346 UA mL-1 , respectively), and was inactive against L. monocytogenes.

Morfometría ultraestructural de la glándula adrenal de ratones infectados experimentalmente con dos aislados venezolanos de trypanosoma evansi / Ultrastructural morphometry of the adrenal gland of mice experimentally infected with two Venezuelan isolates of trypanosoma evansi

Este trabajo describe cualitativamente y analiza cuantitativamente las variaciones ultraestructurales de la glándula adrenal de ratones durante el desarrollo de infecciones murinas experimentales por dos aislados venezolanos de Trypanosoma evansi, uno proveniente de asno y otro de caballo. Las modificaciones submicroscópicas observadas incluyeron alteraciones en el retículo endoplasmático liso, mitocondrias, gotas lipídicas, aparato de Golgi y núcleo. También se apreciaron alteraciones en el número de vesículas de epinefrina, norepinefrina, gránulos de lipofucsina y presencia singular de figuras mielínicas. Estos cambios ultraestructurales estarían asociados a problemas metabólicos que inducirían la muerte de los animales infectados. El estudio cuantitativo demostró diferencias significativas en las características y las magnitudes del daño causado por ambos aislados, siendo el tripanosoma proveniente de caballo más patogénico. El análisis multivariante de los cambios submicroscópicos discriminó la acción patogénica en la glándula adrenal de tripanosomas de la misma especie que provienen de hábitats diferentes (Equus asinus y E. caballus) y de zonas geográficas distintas (estado Apure y estado Guárico).

This work describes qualitatively and analyzes quantitatively the ultrastructural variations of the mice adrenal gland during the development of murine experimental infections by two Venezuelan isolates of Trypanosoma evansi, one de rived from a donkey and another from a horse. The submicroscopic modifications included modifications in the smooth endoplasmic reticulum, mitochondria, lipid droplets, Golgi apparatus, and nucleus. Alterations in the number of epinephrine, norepinephrine vesicles, and lipofuscin granules and singular myelin-like figures were also observed. Such ultrastructural changes would be associated to metabolic problems including the infected animal’s death. The quantitative study demonstrated significant differences in the characteristics and magnitude of the damage caused by both isolates, being more pathogenic the horse derived trypanosome. The multivariate analysis of the submicroscopic changes discriminated the pathogenic action of trypanosomes of the same species coming from different habitats (Equus asinus and E. caballus) and different geographic zones (Apure state and Guarico state) in the adrenal gland.

Nanofabrication of bio-self assembled monolayer and its electrochemical property for toxicant detection.

Cyclic voltammetry (CV) has been used to investigate the electrochemical behavior of a glutathione (GSH) self assembled monolayer on modified gold electrodes (Bio-SAM). The GSH monolayer exhibits an influence on electrode surface activity. Electrochemically immobilized dsDNA onto a Cyt c/GSH-SAM/Au electrode, which is useful for the fabrication of a nanobiosensing device. The immobilized Cyt c followed by dsDNA immobilized films maintained its surface activity and finally dsDNA/Cyt c/GSH-SAM/Au electrode, targeted for the detection of toxicants. The films were characterized by CV, DPV, and AFM. The differential pulse voltammetry (DPV) technique was applied to detect three kinds of common toxins, 2-aminoanthracene (2-AA), 3-bromobenzanthrone (3-BBA) and bisphenol A (BPhA). The electrochemical signals showed good inverse relationship with the increase of concentrations of toxicants. Our proposed system based on electrochemical method with nanoscale film technology can be applied at highly sensitive biosensor for detecting various toxic chemicals.