Evaluation of three recombinant proteins for the development of ELISA and immunochromatographic tests for visceral leishmaniasis serodiagnosis

BACKGROUND Visceral Leishmaniasis (VL) is an infectious disease that is a significant cause of death among infants aged under 1 year and the elderly in Brazil. Serodiagnosis is a mainstay of VL elimination programs; however, it has significant limitations due to low accuracy. OBJECTIVE This study aimed to evaluate three recombinant Leishmania infantum proteins (rFc, rC9, and rA2) selected from previous proteomics and genomics analyses to develop enzyme-linked immunosorbent assay (ELISA) and immunochromatographic tests (ICT) for the serodiagnosis of human VL (HVL) and canine VL (CVL). METHODS A total of 186 human (70 L. infantum-infected symptomatic, 20 other disease-infected, and 96 healthy) and 185 canine (82 L. infantum-infected symptomatic, 27 L. infantum-infected asymptomatic, and 76 healthy) sera samples were used for antibody detection. FINDINGS Of the three proteins, rA2 (91.5% sensitivity and 87% specificity) and rC9 (95.7% sensitivity and 87.5% specificity) displayed the best performance in ELISA-HVL and ELISA-CVL, respectively. ICT-rA2 also displayed the best performance for HVL diagnosis (92.3% sensitivity and 88.0% specificity) and had high concordance with immunofluorescence antibody tests (IFAT), ELISA-rK39, IT-LEISH®, and ELISAEXT. ICT-rFc, ICT-rC9, and ICT-rA2 had sensitivities of 88.6%, 86.5%, and 87.0%, respectively, with specificity values of 84.0%, 92.0%, and 100%, respectively for CVL diagnosis. MAIN CONCLUSIONS The three antigens selected by us are promising candidates for VL diagnosis regardless of the test format, although the antigen combinations and test parameters may warrant further optimisation.

Risk factors of helicobacter pylori infection in an urban community in northeast Brazil and the relationship between the infection and gastric diseases

Abstract INTRODUCTION: Helicobacter pylori, a water contaminant, is the primary pathogenic agent associated with gastric diseases in humans. Exposure to H. pylori is more likely higher in developing countries. This study aimed to evaluate the risk factors associated with H. pylori infection in patients undergoing endoscopy to validate the cause of dyspeptic symptoms in an urban population in northeast Brazil and to compare the urease test and polymerase chain reaction assay results with the histopathological findings. METHODS: We evaluated 200 of 759 individuals with dyspeptic complaints from Campina Grande, State of Paraiba, northeast Brazil. Patients underwent endoscopy, followed by gastric biopsies. Logistic regression analysis was performed to adjust for confounders and to determine significant risk factors of dyspeptic disorders. RESULTS: Women accounted for 72.5% (145/200) of the participants. Approximately 59.8% (120/200) of the samples tested positive for H. pylori based on histological examinations. The specificity of polymerase chain reaction assay was higher than that of the urease test (77% vs. 64%, p=0.034). City drinking water [odds ratio (OR): 2.6; 95% confidence interval (CI): 1.3-5.21; p=0.004] and smoking (OR: 4.0; 95% CI: 1.13-14.5; p=0.031) were the risk factors of H. pylori infection. Belching was the most common symptom associated with H. pylori infection (p=0.05). CONCLUSIONS: The increased risk of H. pylori infection associated with non-treated water consumption indicates the need for improvements in public water treatment and better sanitary conditions because these can be a source of not only H. pylori infections but also other water-borne pathogen infections.

Examining ERBB2 as a candidate gene for susceptibility to leprosy (Hansen’s disease) in Brazil

Leprosy remains prevalent in Brazil. ErbB2 is a receptor for leprosy bacilli entering Schwann cells, which mediates Mycobacterium leprae-induced demyelination and the ERBB2 gene lies within a leprosy susceptibility locus on chromosome 17q11-q21. To determine whether polymorphisms at the ERBB2 locus contribute to this linkage peak, three haplotype tagging single nucleotide polymorphisms (tag-SNPs) (rs2517956, rs2952156, rs1058808) were genotyped in 72 families (208 cases; 372 individuals) from the state of Pará (PA). All three tag-SNPs were associated with leprosy per se [best SNP rs2517959 odds ratio (OR) = 2.22; 95% confidence interval (CI) 1.37-3.59; p = 0.001]. Lepromatous (LL) (OR = 3.25; 95% CI 1.37-7.70; p = 0.007) and tuberculoid (TT) (OR = 1.79; 95% CI 1.04-3.05; p = 0.034) leprosy both contributed to the association, which is consistent with the previous linkage to chromosome 17q11-q21 in the population from PA and supports the functional role of ErbB2 in disease pathogenesis. To attempt to replicate these findings, six SNPs (rs2517955, rs2517956, rs1810132, rs2952156, rs1801200, rs1058808) were genotyped in a population-based sample of 570 leprosy cases and 370 controls from the state of Rio Grande do Norte (RN) and the results were analysed using logistic regression analysis. However, none of the associations were replicated in the RN sample, whether analysed for leprosy per se, LL leprosy, TT leprosy, erythema nodosum leprosum or reversal reaction conditions. The role of polymorphisms at ERBB2 in controlling susceptibility to leprosy in Brazil therefore remains unclear.

Women with cardiovascular risk after preeclampsia: is there follow-up within the Unified Health System in Brazil? / Mulheres com risco cardiovascular apos pre-eclampsia: ha seguimento no Sistema Unico de Saude? / Mujeres con riesgo cardiovascular pos pre-eclampsia: Existe seguimiento en el sistema Unico de Salud?

OBJECTIVES: to identify women with cardiovascular risk, five years after a preeclampsic episode (PE), and identify the follow-up of these women within the Unified Health System (Sistema Único de Saúde - SUS), in the city of Natal/RN. METHODS: a quantitative and exploratory study conducted at the Januário Cicco University Maternity Ward/RN. The sample consisted of 130 women, 65 with a PE episode and 65 who were normotensive. RESULTS: we found statistical significance with regard to body mass index, weight, family history of cardiovascular disease (CVD) and cardiovascular complications when comparing women with previous PE to normotensive women. The groups were unaware of their cardiovascular risk factors and, in addition, they reported difficulties in accessing primary health care (PHC) services. CONCLUSIONS: women with a PE history are at increased risk of developing CVD, unaware of late PE complications, and lacked customized care when compared to normotensive patients. .

OBJETIVOS: identificar mulheres com risco cardiovascular, cinco anos após a pré-eclâmpsia, e averiguar o seguimento dessas mulheres no Sistema Único de Saúde, em Natal, Rio Grande do Norte. MÉTODOS: estudo exploratório, quantitativo, realizado na Maternidade Escola Januário Cicco, em Natal. A amostra foi de 130 mulheres, 65 com pré-eclâmpsia e 65 normotensas. RESULTADOS: constatou-se diferença estatística significativa no que se refere ao índice de massa corpórea, peso, histórico familiar de doenças cardiovasculares e complicação cardiovascular entre as mulheres com pré-eclâmpsia prévia e as normotensas. Os grupos desconheciam os fatores de risco cardiovasculares e, além disso, referiam dificuldades de acesso aos serviços de atenção primária. CONCLUSÕES: as mulheres com histórico de pré-eclâmpsia possuíam risco aumentado de desenvolver doenças cardiovasculares, desconheciam as complicações tardias dessa doença e não recebiam seguimento ambulatorial diferenciado em relação às normotensas. .

OBJETIVOS: identificar mujeres con riesgo cardiovascular, cinco años después de la pre-eclampsia (PE), y averiguar el seguimiento de esas mujeres en el Sistema Único de Salud en Natal/RN. MÉTODOS: estudio exploratorio, cuantitativo, realizado en la Maternidad Escuela Januario Cicco - RN. La muestra fue compuesta de 130 mujeres, 65 con PE y 65 normotensas. RESULTADOS: se constató diferencia estadística significativa en lo que se refiere al índice de masa corpórea, peso, histórico familiar de enfermedades cardiovasculares (ECV) y complicación cardiovascular entre las mujeres con PE previa y las normotensas. Los grupos desconocen los factores de riesgos cardiovasculares y, además de eso, refieren dificultades de acceso en los servicios de atención primaria. CONCLUSIONES: las mujeres con histórico de PE poseen riesgo aumentado de desarrollar ECV, desconocen las complicaciones tardías de esa enfermedad y en relación a las normotensas no reciben seguimiento en ambulatorio especializado. .

Experimental infection parameters in Galea spixii (Rodentia: Caviidae) with Leishmania infantum chagasi

In order to better understand the epidemiological transmission network of leishmaniasis, an endemic disease in Northeast Brazil, we investigated the susceptibility of Spix yellow-toothed cavies (Galea spixii) to the Leishmania infantum chagasi parasite. Nine cavies were experimentally infected, separated into three groups and monitored at 30, 90 and 180 days, respectively. Amastigotes were identified in the spleen slides of two cavies killed 180 days after infection. Antibodies against the L. i. chagasi were identified in one of the cavies. This demonstrates that G. spixii is in fact capable of maintaining a stable infection by L. i. chagasi without alterations in biochemical and hematological parameters of the host and without perceivable micro and macroscopic lesions.

Percepção materna com o nascimento prematuro e vivência da gravidez com pré-eclampsia / Maternal perception of premature birth and the experience of pre-eclampsia pregnancy

OBJETIVO: Analisar a vivência materna diante de gravidez com pré-eclampsia e com recém-nascido prematuro hospitalizado em unidade de terapia intensiva neonatal. MÉTODOS: Estudo qualitativo com a utilização da técnica do grupo focal, realizado com 28 mulheres em maternidade referência para gestações de risco no Rio Grande do Norte, em 2004. As mães incluídas tiveram gravidez com pré-eclampsia e parto pré-termo e conseqüente internação do recém-nascido em Unidade de Terapia Intensiva Neonatal. Os dados foram analisados por meio da técnica análise de conteúdo temática categorial, abordando três núcleos temáticos: informações sobre a pré-eclampsia no pré-natal; vivências com o recém-nascido pré-termo; e a percepção das usuárias sobre o atendimento dos profissionais. RESULTADOS: Os relatos maternos evidenciaram o desconhecimento das participantes sobre a pré-eclampsia e sua associação com a prematuridade. Foram detectadas dificuldades inerentes à definição do papel materno nos cuidados com o filho, acentuadas pelas falhas na relação entre os profissionais de saúde e os usuários. CONCLUSÕES: Parte das dificuldades vividas pelas mães no contexto da pré-eclampsia e da prematuridade foram agravadas pela ausência de informação ou inadequação para o entendimento das usuárias. Os resultados mostram que dificuldades vividas por mães no contexto da pré-eclampsia e da prematuridade são agravadas pela ausência de informação ou inadequação para o seu entendimento.

OBJECTIVE: To analyze maternal experiences of preeclampsia pregnancy with premature birth at a neonatal intensive care unit. METHODS: A qualitative study using the focus group technique was conducted with 28 women in a facility specialized in high-risk pregnancies in the state of Rio Grande do Norte, Northeastern Brazil, in 2004. Mothers included had had preeclampsia during pregnancy and a preterm delivery with consequent hospitalization of their baby at a neonatal intensive care unit. The data were analyzed using thematic content analysis of three thematic nuclei subjects: information about preeclampsia during prenatal care; experiences with the preterm newborn, and their perception of neonatal intensive care unit professionals' attitudes. RESULTS: Maternal reports showed subjects' lack of knowledge with regard to preeclampsia and its association with prematurity. Difficulties inherent to the maternal role of caring for the child in the neonatal intensive care unit were identified, accentuated by communication flaws between health professionals and users. CONCLUSIONS: Some difficulties experienced by the mothers, in the context of preeclampsia and prematurity, were aggravated by lack or inadequacy of information provided to the users.

Síndrome HELLP: considerações acerca de diagnóstico e conduta / Hellp syndrome: considerations about diagnosis and management

A síndrome HELLP é uma grave complicação da pré-eclampsia, cursando com elevada morbimortalidade materna e perinatal. Controvérsias ocorrem com relação à incidência, critérios de diagnóstico e conduta nesta síndrome. As mortes perinatais estão relacionadas com o descolamento prematuro da placenta, asfixia intra-útero e prematuridade. A utilização de altas doses de corticóide no tratamento parece proporcionar melhores resultados maternos e perinatais, no entanto, são necessários estudos randomizados para sua completa e segura utilização na síndrome HELLP. O maior desafio diante desta síndrome é o diagnóstico precoce, intervenção oportuna e a prevenção das complicações. Sendo assim, as pacientes com suspeita de síndrome HELLP deverão ser encaminhadas para um serviço de assistência terciária para que sejam assistidas por uma equipe especializada, e tão logo possível, promova-se a interrupção da gestação.

Bacterial translocation in rats nonfunctioning diverted distal colon

PURPOSE: To investigate whether the alterations of the diverted colon segment mucosa, evidenced in fecal colitis, would be able to alter Bacterial Translocation (BT). METHODS: Sixty-two Wistar male rats ranging from 220 to 320 grams of weight, were divided in two groups: A (Colostomy) and B (Control), with 31 animals each one. In group A, all animals underwent end colostomy, one stoma, in ascending colon; and in the 70th POD was injected in five rats, by rectal route diverted segment - 2ml of a 0.9 percent saline solution in animals (A1 subgroup); in eight it was inoculated, by rectal route, 2ml of a solution containing Escherichia coli ATCC 25922 (American Type Culture Collection), in a concentration of 10(8) Colony Forming Unit for milliliters (CFU/ml) - A2 Subgroup; in ten animals the same solution of E. coli was inoculated, in a concentration of 10(11) CFU/ml (A3 Subgroup); and in eight it was collected part of the mucus found in the diverted distal colonic segment for neutral sugars and total proteins dosage (A4 subgroup). The animals from the group B underwent the same procedures of group A, but with differences in the colostomy confection. In rats from subgroups A1, A2, A3, B1, B2, and B3 2ml of blood were aspirated from the heart, and fragments from mesenteric lymphatic nodule, liver, spleen, lung and kidney taken for microbiological analysis, after their death. This analysis consisted of evidencing the presence of E. coli ATCC 25922 CFU. Mann-Whitney and ANOVA Tests were applied as analytic techniques for association of variables. RESULTS: The occurrence of BT was evidenced only in those animals in which inoculated concentration of E. coli ATCC 25922, reached levels of 10(11)CFU/ml, i.e. in Subgroups A3 and B3, although, being significantly greater (80 percent) in those animals without colostomy (subgroup B3) when compared to the ones with colostomy (20 percent) from the subgroup A3 (P <0.05). Lung, liver and mesenteric lymphatic...

OBJETIVO: Investigar se as alterações do cólon desfuncionalizado, evidenciadas na colite de derivação fecal, seriam capazes de permitir Translocação Bacteriana (TB), ou se a mucosa intestinal atrofiada permitiria a passagem de bactérias para órgãos à distância. MÉTODOS: Foram utilizados 62 ratos Wistar, machos, pesando entre 220 e 320 gramas, divididos em dois grupos: A (Colostomia) e B (Controle), contendo cada um 31 animais. No grupo A, os animais foram submetidos à colostomia, terminal boca única, em cólon ascendente. A partir do 70° dia de observação os seguintes procedimentos foram adotados: em cinco ratos foi injetado por via retal no segmento desfuncionalizado - 2ml de uma solução salina 0,9 por cento nos animais (subgrupo A1); em oito inoculou-se, por via retal, uma solução de 2ml contendo Escherichia coli ATCC 25922 (American Type Culture Collection), na concentração de 10(8) Unidades Formadoras de Colônias por mililitros (UFC/ml) - Subgrupo A2; em dez animais inoculava-se a mesma solução de E. coli, na concentração de 10(11)UFC/ml (Subgrupo A3); e em oito colhia-se o muco do segmento colônico distal desfuncionalizado, para dosagens de açúcares neutros e proteínas totais (subgrupo A4). Os animais do grupo B foram submetidos aos mesmos procedimentos do grupo A, e não foram submetidos à colostomia. Nos animais dos subgrupos A1, A2, A3, B1, B2, e B3, após serem mortos, realizou-se punção cardíaca para coleta de 2ml de sangue e retiraram-se fragmentos de tecidos de linfonodo do mesocólon, fígado, baço, pulmão e rim, para análise microbiológica. Essa análise consistia em evidenciar a presença de UFC de Escherichia coli ATCC 25922. Os testes de Mann-Whitney e Anova foram aplicados como técnicas investigativas para associação das variáveis. RESULTADOS: A ocorrência de TB, só foi evidenciada nos animais em que a concentração inoculada de Escherichia coli ATCC 25922, atingia níveis de 10(11)UFC/ml, ou seja, nos Subgrupos A3 e B3, no entanto,...

Characteristics of the bological cycle of Lutzomyia evandroi Costa Lima & Antunes, 1936 (Diptera: Psychodidae) under experimental conditions

Lutzomyia evandroi Costa Lima and Antunes, 1936 is found in Rio Grande do Norte, northeastern Brazil, in areas of visceral and mucocutaneous leishmaniasis and follows the same geographic distribution of L. longipalpis. The biological cycle, oviposition, morphological and behavioral characteristics of the species were studied under experimental conditions. The average number of eggs per wild caught female varied from 21 to 50 eggs along the year, with a peak occurring between January and March and another in August, with oviposition lasting for 4 to 12 days. The mean larval phase was 24 days. Ovipositing rates were influenced by rainfall and temperature indexes, with an increase of eggs per oviposition at the beginning and at the end of the rainy season, and a decrease at the peak of the rainy season