In-house ELISA method to analyze anti-Trypanosoma cruzi IgG reactivity for differential diagnosis and evaluation of Chagas disease morbidity

INTRODUCTION: The goal was to develop an in-house serological method with high specificity and sensitivity for diagnosis and monitoring of Chagas disease morbidity. METHODS: With this purpose, the reactivities of anti-T. cruzi IgG and subclasses were tested in successive serum dilutions of patients from Berilo municipality, Jequitinhonha Valley, Minas Gerais, Brazil. The performance of the in-house ELISA was also evaluated in samples from other relevant infectious diseases, including HIV, hepatitis C (HCV), syphilis (SYP), visceral leishmaniasis (VL), and American tegumentary leishmaniasis (ATL), and noninfected controls (NI). Further analysis was performed to evaluate the applicability of this in-house methodology for monitoring Chagas disease morbidity into three groups of patients: indeterminate (IND), cardiac (CARD), and digestive/mixed (DIG/Mix), based on their clinical status. RESULTS: The analysis of total IgG reactivity at serum dilution 1:40 was an excellent approach to Chagas disease diagnosis (100% sensitivity and specificity). The analysis of IgG subclasses showed cross-reactivity, mainly with NI, VL, and ATL, at all selected serum dilutions. Based on the data analysis, the IND group displayed higher IgG3 levels and the DIG/Mix group presented higher levels of total IgG as compared with the IND and CARD groups. CONCLUSIONS: These findings demonstrated that methodology presents promising applicability in the analysis of anti-T. cruzi IgG reactivity for the differential diagnosis and evaluation of Chagas disease morbidity.

Production and diagnostic application of recombinant domain III of West Nile envelope protein in Brazil

INTRODUCTION: West Nile virus (WNV) is a flavivirus with a natural cycle involving mosquitoes and birds. Over the last 11 years, WNV has spread throughout the Americas with the imminent risk of its introduction in Brazil. METHODS: Envelope protein domain III of WNV (rDIII) was bacterially expressed and purified. An enzyme-linked immunosorbent assay with WNV rDIII antigen was standardized against mouse immune fluids (MIAFs) of different flavivirus. RESULTS: WNV rDIII reacted strongly with St. Louis encephalitis virus (SLEV) MIAF but not with other flaviviruses. CONCLUSIONS: This antigen may be a potentially useful tool for serologic diagnosis and may contribute in future epidemiological surveillance of WNV infections in Brazil.

Molecular and biological characterization of Trypanosoma cruzi strains isolated from children from Jequitinhonha Valley, State of Minas Gerais, Brazil

Introduction The biological diversity of Trypanosoma cruzi strains plays an important role in the clinical and epidemiological features of Chagas disease. Methods Eight T. cruzi strains isolated from children living in a Chagas disease vector-controlled area of Jequitinhonha Valley, State of Minas Gerais, Brazil, were genetically and biologically characterized. Results The characterizations demonstrated that all of the strains belonged to T. cruzi II, and showed high infectivity and a variable mean maximum peak of parasitemia. Six strains displayed low parasitemia, and two displayed moderate parasitemia. Later peaks of parasitemia and a predominance of intermediate and large trypomastigotes in all T. cruzi strains were observed. The mean pre-patent period was relatively short (4.2±0.25 to 13.7±3.08 days), whereas the patent period ranged from 3.3±1.08 to 34.5±3.52 days. Mortality was observed only in animals infected with strain 806 (62.5%). Histopathological analysis of the heart showed that strains 501 and 806 caused inflammation, but fibrosis was observed only in animals infected with strain 806. Conclusions The results indicate the presence of an association between the biological behavior in mice and the genetic characteristics of the parasites. The study also confirmed general data from Brazil where T. cruzi II lineage is the most prevalent in the domiciliary cycle and generally has low virulence, with some strains capable of inducing inflammatory processes and fibrosis.

Performance of a rapid diagnostic test for the detection of visceral leishmaniasis in a large urban setting

Introduction Rapid diagnostic tests (RDTs) may improve the early detection of visceral leishmaniasis (VL), but their real-world performance requires additional study. Therefore, we evaluated the performance of an rK39-based RDT (Kalazar Detect™) for the detection of VL in an endemic, large urban area. Methods Data were collected from a registry of rK39 RDT performed at 11 emergency care units in Belo Horizonte, Brazil, and from a national database of reportable communicable diseases of the Sistema de Informação de Agravos de Notificação (SINAN). Results The rapid rK39 test was performed in 476 patients, with 114 (23.9%) positive results. The analysis of rK39 RDT performance was based on 381 (80%) cases reported to the SINAN database, of which 145 (38.1%) were confirmed cases. Estimates for sensitivity and specificity were 72.4% (95% CI: 64.6-79%) and 99.6% (95%CI: 97.6-99.9%), respectively. Positive and negative predictive values were estimated at 99.1% (95%CI: 94.9-99.8%) and 85.5% (95%CI: 80.8-89.1%), respectively. In addition, close agreement between the rK39 RDT and indirect immunofluorescence was observed. Conclusions In summary, the rK39 RDT showed a high specificity but only moderate sensitivity. In endemic areas for VL, treatment may be considered in cases with clinical manifestations and a positive rK39 RDT, but those with a negative test should be subjected to further investigation.

Occurrence of potentially pathogenic Vibrio in oysters (Crassostrea gigas) and waters from bivalve mollusk cultivations in the South Bay of Santa Catarina

Introduction This research aimed to identify and quantify potentially pathogenic Vibrio from different cultivations of bivalve shellfish in the State of Santa Catarina, Brazil, and water regions in the South Bay, as well as correlate the incidence of these microorganisms with the physicochemical parameters of marine waters. Methods Between October 2008 and March 2009, 60 oyster and seawater samples were collected from six regions of bivalve mollusk cultivation, and these samples were submitted for Vibrio counts. Results Twenty-nine (48.3%) oyster samples were revealed to be contaminated with one or more Vibrio species. The Vibrio parahaemolyticus and Vibrio vulnificus counts in the samples ranged from < 0.5 log10 Most Probable Number (MPN) g–1 to 2.3 log10 MPN g–1 oyster and from < 0.5 log10 MPN g–1 to 2.1 log10 MPN g–1 oyster, respectively. Of the 60 seawater samples analyzed, 44 (73.3%) showed signs of contamination with one or more vibrio species. The counts of V. parahaemolyticus and V. vulnificus in the samples ranged from < 0.3 log10 MPN·100mL–1 to 1.7 log10MPN·100mL–1 seawater and from < 0.3 log10 MPN·100mL–1 to 2.0 log10 MPN·100mL–1 seawater, respectively. A positive correlation between V. vulnificus counts and the seawater temperature as well as a negative correlation between the V. parahaemolyticus counts and salinity were observed. Conclusions The results suggest the need to implement strategies to prevent vibrio diseases from being transmitted by the consumption of contaminated bivalve shellfish.

Expression of annexin A1 in Leishmania-infected skin and its correlation with histopathological features

ABSTRACTINTRODUCTION:The aim of this study was quantify annexin A1 expression in macrophages and cluster of differentiation 4 (CD4) + and cluster of differentiation 8 (CD8)+ T cells from the skin of patients with cutaneous leishmaniasis (n=55) and correlate with histopathological aspects.METHODS:Infecting species were identified by polymerase chain reaction-restriction fragment length polymorphism, and expression of annexin A1 was analyzed by immunofluorescence.RESULTS:All patients (n = 55) were infected with Leishmania braziliensis . Annexin A1 was expressed more abundantly in CD163 + macrophages in infected skin (p < 0.0001) than in uninfected skin. In addition, macrophages in necrotic exudative reaction lesions expressed annexin A1 at higher levels than those observed in granulomatous (p < 0.01) and cellular lesions p < 0.05). This difference might be due to the need to clear both parasites and necrotic tissue from necrotic lesions. CD4 + cells in cellular lesions expressed annexin A1 more abundantly than did those in necrotic (p < 0.05) and granulomatous lesions (p < 0.01). Expression in CD8 + T cells followed the same trend. These differences might be due to the pervasiveness of lymphohistiocytic and plasmacytic infiltrate in cellular lesions.CONCLUSIONS:Annexin A1 is differentially expressed in CD163 + macrophages and T cells depending on the histopathological features of Leishmania -infected skin, which might affect cell activation.

Vanamea GEN. NOV. FOR Livoneca symmetricaVAN NAME,1925, (CRUSTACEA, ISOPODA, CYMOTHOIDAE) AND A REDESCRIPTION OF THE SPECIES BASED ON SPECIMENS FROM BRAZILIAN PIRANHAS

Studies of the cymothoid isopod Livoneca symmetricaVan Name, 1925, showed that this species has characters that preclude its inclusion in LivonecaLeach, 1818, or in any other known genus. The species is redescribed on the basis of male and female specimens from the mouth cavities of Amazonian piranhas (Serrasalmus spilopleura(Kner) and S. elongatusKner) and Vanameagen. nov. is proposed for it. The new genus is defined as having: a cephalon that is not immersed in pereonite 1; mandibles that are "foot-shaped" and without incisors, pereopods that are long and unequal in size and shape; a pleon that is not immersed in the pereon; and a pleotelson that is inflated anteriorly and medially.

New records of Fredius denticulatus (H. Milne-Edwards, 1853) and F. reflexifrons (Ortmann, 1897), and the eastern limits of the distribution of pseudothelphusid crabs (Crustacea: Decapoda) in Brazil

The occurence of Fredius reflexifrons (Ortmann, 1897), a pseudothelphusid crab widely distributed in the Amazon region and the Atlantic Guianas, is recorded from the state of Ceará, northeastern Brazil. Other records of this species and Fredius denticulatus (H. Milne-Edwards, 1853) from the Amazon region are also presented. A discussion is made on the eastern limits of the distribution of the family Pseudothelphusidae in Brazil.

Inventário de Depressão de Beck (BDI) e Escala de Avaliação de Depressão de Hamilton (HAM-D) em pacientes com epilepsia

OBJETIVO: Determinar os pontos de melhor sensibilidade e especificidade do Inventário de Depressão de Beck (BDI) e da Escala de Avaliação de Depressão de Hamilton (HAM-D) no diagnóstico de depressão associada à epilepsia. MÉTODOS: Setenta e três pacientes de um centro de referência no tratamento da epilepsia foram submetidos à avaliação neuropsiquiátrica. Foram colhidos dados clínicos e sociodemográficos, sendo utilizados os seguintes instrumentos: entrevista clínica estruturada (MINI-PLUS) para diagnóstico psiquiátrico conforme o DSM-IV, HAM-D e BDI. RESULTADOS: No momento da entrevista, 27,4% dos pacientes estavam deprimidos e 37% preenchiam critérios para diagnóstico de depressão maior ao longo da vida. A análise da curva ROC indicou que o ponto de corte em 16 (> 16) para o BDI (sensibilidade de 94,4%, especificidade de 90,6%) e em 16 (> 16) para a HAM-D (sensibilidade de 95%, especificidade de 75,5%) representou dicotomização ótima entre deprimidos e não deprimidos. Ambos os instrumentos apresentaram um valor preditivo negativo superior a 95%. CONCLUSÃO: A frequência de depressão maior é elevada em pacientes com epilepsia. BDI e a HAM-D podem auxiliar o clínico na identificação da depressão associada à epilepsia, diminuindo seu subdiagnóstico.

Escala Hamilton: estudo das características psicométricas em uma amostra do sul do Brasil

Objetivo Investigar as características psicométricas de uma versão traduzida da escala, propondo uma Versão Revisada que atenda aos critérios de adaptação transcultural para o contexto brasileiro. Métodos Este estudo incluiu 231 sujeitos – deprimidos (45,5%), bipolares (7,8%) e saudáveis (46,7%) – que participaram de uma pesquisa epidemiológica no sul do Brasil. A avaliação de transtornos mentais foi realizada por meio da Clinical Interview for DSM-IV (SCID) e uma versão traduzida da Escala de Avaliação de Depressão de Hamilton (HAM-D), que habitualmente vem sendo utilizada no país sem estudos de adaptação. Resultados Identificou-se o ponto de corte (9 pontos) para discriminar a presença ou não de sintomas de depressão pela análise da curva ROC, resultando em uma sensibilidade e especificidade de 90 e 91%, respectivamente. A validade interna foi investigada pela análise fatorial e consistência dos itens. Dos 17 itens originais, apenas o item que avalia a “consciência do transtorno” não apresentou carga fatorial satisfatória para avaliar depressão geral e foi eliminado; os 16 restantes agruparam-se em cinco dimensões, denominadas: Humor deprimido, Anorexia, Insônia, Somatização e Ansiedade, as quais, com exceção da última, mostraram homogeneidade nos seus construtos (coeficientes alfa entre 0,66 e 0,78). Na análise de conteúdo dos itens, cinco especialistas sugeriram alterações redacionais em sete itens. Conclusão O estudo determina um ponto de corte diferente do original e evidencia características psicométricas favoráveis para a utilização da escala no Brasil.

Narrative of the expedition of an American squadron to the China Seas and Japan : performed in the years 1852, 1853, and 1854, under the command of Commodore M.C. Perry, United States Navy, by order of the Government of the United States /

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Anatomia histológica de alguns órgãos da lagarta e do adulto da Brassolis sophorae laurentii Stichel, 1925

O autor continuando os seus estudos sôbre a Brassolis sophorae laurentii Stichel, estende-o a anatomia histológica de alguns órgãos, do adulto e da lagarta, especialmente do aparelho digestivo e do reprodutor.

Anatomia morfológica, externa e interna da lagarta, e do adulto da Brassolis sophorae laurentii Stichel, 1925

O autor neste trabalho faz a descrição morfológica externa e interna da lagarta e do adulto de Brassolis sophorae Laurentii Stichel, inseto praga das palmeiras. Descreve o aspecto externo e após dissecação em líquido Ringer, estuda a morfologia dos diversos aparelhos, inclusive a genitalia do macho e ovos.