Rapid and Highly Sensitive DNA Flow Technology Platform to Detect Tick-Borne Bacterial Pathogens in Clinical Samples.

Zoonotic diseases represent a significant public health concern worldwide due to the emergence/re-emergence of vector-borne diseases in the last decade. Ticks are the most important vectors in the northern hemisphere and can transmit diseases such as Lyme disease, human granulocytic anaplasmosis, and spotted fever rickettsioses, among others. Therefore, there is a growing need to develop better and faster diagnostic tools that can detect zoonotic human pathogens in clinical samples. In this study, we present the results for a new kit tick-borne bacteria flow chip (TBFC), which allows the simultaneous screening of seven different bacterial pathogens in human samples using a DNA flow technology platform (hybriSpot system). The analytical sensitivity and specificity of the TBFC were calculated spiking bacterial DNA in human DNA samples, and the results were compared with an in-house single PCR-reverse line blot (RLB) routinely used for diagnosis at the National Center for Microbiology in Spain. The analytical sensitivity and specificity of the TBFC were almost identical to the PCR-RLBs used in diagnosis. In addition, samples from patients (n = 212) with a wide range of clinical signs/symptoms consistent with multisystem disorders suggestive of a tick-borne infection were tested using the TBFC, and the results were compared with those obtained by PCR-RLB. The concordance of both methods using patient samples was 97.2%. The TBFC kit is a rapid new and cost-efficient diagnostic molecular tool capable of detecting tick-borne pathogens in clinical samples.

Distribution of Bartonella henselae variants in patients, reservoir hosts and vectors in Spain.

We have studied the diversity of B. henselae circulating in patients, reservoir hosts and vectors in Spain. In total, we have fully characterized 53 clinical samples from 46 patients, as well as 78 B. henselae isolates obtained from 35 cats from La Rioja and Catalonia (northeastern Spain), four positive cat blood samples from which no isolates were obtained, and three positive fleas by Multiple Locus Sequence Typing and Multiple Locus Variable Number Tandem Repeats Analysis. This study represents the largest series of human cases characterized with these methods, with 10 different sequence types and 41 MLVA profiles. Two of the sequence types and 35 of the profiles were not described previously. Most of the B. henselae variants belonged to ST5. Also, we have identified a common profile (72) which is well distributed in Spain and was found to persist over time. Indeed, this profile seems to be the origin from which most of the variants identified in this study have been generated. In addition, ST5, ST6 and ST9 were found associated with felines, whereas ST1, ST5 and ST8 were the most frequent sequence types found infecting humans. Interestingly, some of the feline associated variants never found on patients were located in a separate clade, which could represent a group of strains less pathogenic for humans.

Molecular method for the characterization of Coxiella burnetii from clinical and environmental samples: variability of genotypes in Spain.

BACKGROUND: Coxiella burnetii is a highly clonal microorganism which is difficult to culture, requiring BSL3 conditions for its propagation. This leads to a scarce availability of isolates worldwide. On the other hand, published methods of characterization have delineated up to 8 different genomic groups and 36 genotypes. However, all these methodologies, with the exception of one that exhibited limited discriminatory power (3 genotypes), rely on performing between 10 and 20 PCR amplifications or sequencing long fragments of DNA, which make their direct application to clinical samples impracticable and leads to a scarce accessibility of data on the circulation of C. burnetii genotypes. RESULTS: To assess the variability of this organism in Spain, we have developed a novel method that consists of a multiplex (8 targets) PCR and hybridization with specific probes that reproduce the previous classification of this organism into 8 genomic groups, and up to 16 genotypes. It allows for a direct characterization from clinical and environmental samples in a single run, which will help in the study of the different genotypes circulating in wild and domestic cycles as well as from sporadic human cases and outbreaks. The method has been validated with reference isolates. A high variability of C. burnetii has been found in Spain among 90 samples tested, detecting 10 different genotypes, being those adaA negative associated with acute Q fever cases presenting as fever of intermediate duration with liver involvement and with chronic cases. Genotypes infecting humans are also found in sheep, goats, rats, wild boar and ticks, and the only genotype found in cattle has never been found among our clinical samples. CONCLUSIONS: This newly developed methodology has permitted to demonstrate that C. burnetii is highly variable in Spain. With the data presented here, cattle seem not to participate in the transmission of C. burnetii to humans in the samples studied, while sheep, goats, wild boar, rats and ticks share genotypes with the human population.

Case report: melioidosis imported from West Africa to Europe.

We report the first case of imported melioidosis in Spain from a diabetic immigrant who visited West Africa during the rainy season. Because of the unusual presentation of this disease in Africa, clinical and microbiological diagnosis of imported melioidosis from this continent can be very elusive.

Análise das lesões intra-articulares na instabilidade anterior do ombro / Analysis of intra-articular lesions in anterior shoulder instability

Introdução: Instabilidade anterior do ombro é uma afecção frequente, na qual uma de suas consequências é o surgimento de lesões labrais, ósseas e do manguito rotador. Estas lesões que, muitas vezes, estão associadas, variam no seu tamanho de acordo com a causa e número de episódios de luxações. Objetivo: Descrever as alterações intra-articulares encontradas nos pacientes submetidos ao tratamento artroscópico para instabilidade anterior do ombro e correlacionar os achados com a causa da luxação e o número de episódios até o tratamento. Material e métodos: Entre agosto de 2006 e julho de 2007, 60 cirurgias para instabilidade anterior do ombro foram realizadas no Instituto Nacional de Traumatologia e Ortopedia (INTO). Protocolo pré-operatório específico identificando-se: idade, sexo, mecanismo do primeiro episódio de luxação (traumática ou atraumática) e número de luxações foi preenchido em todos os casos. Ao final do procedimento cirúrgico foi preenchido um protocolo descrevendo a eventual presença das seguintes lesões: Bankart, Hill-Sachs, SLAP, ALPSA, HAGL, ruptura do manguito rotador e lesão óssea da glenoide. As lesões da glenoide foram divididas em três tipos: lesão condral sem defeito ósseo, lesões com perda óssea menor e maior que 27%. Resultados: Dos 60 pacientes, 42 eram do sexo masculino e 18 do feminino. A média de idade foi de 30 anos (16-54), 45 casos traumáticos e 15 atraumáticos e a média de luxações foi de 20 episódios (1-100). A lesão de Bankart esteve presente em 57 pacientes (95%), Hill-Sachs em 53 (88%), a do manguito rotador em três casos (5%), SLAP em 11 (18%) e a lesão ALPSA em três (5%). Conclusão: Todos os casos traumáticos e a maioria dos atraumáticos apresentaram lesão do labrum anterior. Não houve associação entre o número de luxações e lesão óssea na glenoide ou entre o mecanismo do primeiro episódio e presença de lesão óssea.

Molecular method for discrimination between Francisella tularensis and Francisella-like endosymbionts.

Environmental studies on the distribution of Francisella spp. are hampered by the frequency of Francisella-like endosymbionts that can produce a misleading positive result. A new, efficient molecular method for detection of Francisella tularensis and its discrimination from Francisella-like endosymbionts, as well as two variants associated with human disease (unusual F. tularensis strain FnSp1 and F. tularensis subsp. novicida-like strain 3523), is described. The method is highly specific and sensitive, detecting up to one plasmid copy or 10 genome equivalents.

Molecular method for identification of Rickettsia species in clinical and environmental samples.

We present a PCR method targeting the 23S-5S internal transcribed spacer coupled with reverse line blotting that allows Rickettsia species detection and identification in a single step. The method is highly sensitive and specific in identifying Rickettsia species from both patient and environmental samples. The generic approach used allowed us to identify new pathogens.

Prevenção de hematoma subgaleal pós-operatório. técnica de curativo / New technique for postoperative craniotomy dressing

Uma das complicações que ocorrem no pós-operatório das craniotomias é a formação de coleções subgaleais por extravasamento de sangue ao nível da incisão. Apesar da utilização de curativos compressivos, esses fatos ocorrem em grande número de casos e, em alguns serviços, são utilizados drenos externos para tentar minimizar esse problema. Como, além de onerar o procedimento cirúrgico, essa técnica ainda possibilita o aparecimento de infecção, o Serviço de Neurocirurgia do Hospital Universitário do Norte do Paraná passou a utilizar uma nova técnica de curativo, em que são utilizadas luva e dreno de Penrose parcialmente cheios de soro, respectivamente sobre a craniotomia e na região supra-orbitária, conseguindo evitar essa complicação.

Valores de referencia para plumbemia em uma populacao urbana do Sul do Brasil / Reference values for lead in blood in an urban population in southern Brazil

Objetive. To describe the reference values for lead in blood in an urban population in the city of Londrina, in the state of Paraná, Brasil. Materials and methods. The reference population was composed of 520 adult volunteers who were assessed from November 1994 to December 1996. Exclusion criteria were: occupational exposure to lead, exposure through personal habits or practice, smoking more than 10 cigarettes per day, and living near indistrial plants or other places that use lead in the production processes. Also excluded were individuals with abnormal clical or laboratory results or with chonic diseases or cardiovascular disorders. Lead blood levels were determined using air-acetylene flame atomic absorption spectrophotometry. The detectable limit was 1.23ug/dL. After the analyses of lead in blood, the following values were determined: minimum value, first quartile, median, third quartile, and maximum value; geometric mean, 95 confidence interval; experimental interval; and reference value. Results. The reference values for lead in blood ranger from 1.20ug/dL to 13.72ug/dL. The geometric mean was 5.5 ug/dL. Conclusions. In general, the values found in this study are lower than those that have been reported for other countries. Additional date should be gathered from Brazilian populations living in more-industrialized areas

Valores de referência para plumbemia em uma população urbana do Sul do Brasil / Reference values for lead in blood in an urban population in southern Brazil

Objetivo. Descrever os valores de referência para chumbo em sangue em uma população urbana do Município de Londrina, Estado do Paraná, Brasil. Métodos. A população de referência foi constituída por 520 voluntários adultos avaliados de novembro de 1994 a dezembro de 1996. Os critérios de exclusão foram: exposição ocupacional ao chumbo, exposição através de hábitos, fumar mais de 10 cigarros por dia e morar perto de indústrias ou de locais que utilizam o chumbo em seus processos produtivos. Também foram excluídos indivíduos com valores de exames clínicos e laboratoriais fora da normalidade, doenças crônicas e distúrbios cardiovasculares. Os teores de chumbo em sangue foram determinados por espectrofotometria de absorção atômica com chama de ar-acetileno. O limite de detecção obtido foi 1,23 mig/dL. Após as análises de chumbo em sangue, foram determinados valor mínimo, primeiro quartil, mediana, terceiro quartil e valor máximo; média geométrica; intervalo de confiança de 95 por cento; intervalo experimental; e valor de referência. Resultados. Os valores de referência para plumbemia variaram de 1,20 a 13,72 mig/dL. A média geométrica foi igual a 5,5 mig/dL. Conclusões. Em geral, os valores obtidos no presente estudo são mais baixos do que aqueles obtidos em outros países. Estudos adicionais deveriam enfocar a obtenção de valores de referência em outras populações brasileiras que vivem em regiões mais industrializadas.

Objective. To describe the reference values for lead in blood in an urban population in the city of Londrina, in the state of Paraná, Brazil. Materials and methods. The reference population was composed of 520 adult volunteers who were assessed from November 1994 to December 1996. Exclusion criteria were: occupational exposure to lead, exposure through personal habits or practices, smoking more than 10 cigarettes per day, and living near industrial plants or other places that use lead in their production processes. Also excluded were individuals with abnormal clinical or laboratory results or with chronic diseases or cardiovascular disorders. Lead blood levels were determined using air-acetylene flame atomic absorption spectrophotometry. The detectable limit was 1.23 mug/dL. After the analyses of lead in blood, the following values were determined: minimum value, first quartile, median, third quartile, and maximum value; geometric mean; 95 percent confidence interval; experimental interval; and reference value. Results. The reference values for lead in blood ranged from 1.20 mug/dL to 13.72 mug/ dL. The geometric mean was 5.5 mug/dL. Conclusions. In general, the values found in this study are lower than those that have been reported for other countries. Additional data should be gathered from Brazilian populations living in more-industrialized areas.

A Faculdade dos meus dias / The College of my time

Trata-se de um relato autobiográfico que tem por objetivo näo manter no esquecimento fatos que contribuíram para enaltecer uma casa de ensino - a Faculdade de Medicina do Rio de Janeiro - que tanto serviu ao país. Reflete os fatos e emoçöes retidas na mente do autor, salvo pelos dados referentes a antigos professores, que foram tirados do livro - O Centenário da Faculdade de Medicina', de autoria de Fernando de Magalhäes.(MAM)

A Faculdade dos meus dias / The College of my time

Trata-se de um relato autobiográfico que tem por objetivo näo manter no esquecimento fatos que contribuíram para enaltecer uma casa de ensino - a Faculdade de Medicina do Rio de Janeiro - que tanto serviu ao país. Reflete os fatos e emoçöes retidas na mente do autor, salvo pelos dados referentes a antigos professores, que foram tirados do livro - O Centenário da Faculdade de Medicina', de autoria de Fernando de Magalhäes.(MAM)