SEROPOSITIVITY FOR ASCARIOSIS AND TOXOCARIOSIS AND CYTOKINE EXPRESSION AMONG THE INDIGENOUS PEOPLE IN THE VENEZUELAN DELTA REGION

The present study aimed at measuring seropositivities for infection by Ascaris suum and Toxocara canis using the excretory/secretory (E/S) antigens from Ascaris suum (AES) and Toxocara canis (TES) within an indigenous population. In addition, quantification of cytokine expressions in peripheral blood cells was determined. A total of 50 Warao indigenous were included; of which 43 were adults and seven children. In adults, 44.1% were seropositive for both parasites; whereas children had only seropositivity to one or the other helminth. For ascariosis, the percentage of AES seropositivity in adults and children was high; 23.3% and 57.1%, respectively. While that for toxocariosis, the percentage of TES seropositivity in adults and children was low; 9.3% and 14.3%, respectively. The percentage of seronegativity was comparable for AES and TES antigens in adults (27.9%) and children (28.6%). When positive sera were analyzed by Western blotting technique using AES antigens; three bands of 97.2, 193.6 and 200.2 kDas were mostly recognized. When the TES antigens were used, nine major bands were mostly identified; 47.4, 52.2, 84.9, 98.2, 119.1, 131.3, 175.6, 184.4 and 193.6 kDas. Stool examinations showed that Blastocystis hominis, Hymenolepis nana and Entamoeba coli were the most commonly observed intestinal parasites. Quantification of cytokines IFN-&#947;, IL-2, IL-6, TGF-β, TNF-α, IL-10 and IL-4 expressions showed that there was only a significant increased expression of IL-4 in indigenous with TES seropositivity (p < 0.002). Ascaris and Toxocara seropositivity was prevalent among Warao indigenous.

Implicações clínicas e imunológicas da associação entre o HTLV-1 e a estrongiloidíase

A estrongiloidíase é uma das mais importantes helmintíases em países tropicais e estudos epidemiológicos têm demonstrado associação desta parasitose com o vírus HTLV-1. Em regiões onde estes dois agentes são endêmicos a coinfecção pode resultar no desenvolvimento de formas disseminadas da estrongiloidíase assim como em estrongiloidíase recorrente. Enquanto que o vírus HTLV-1 está relacionado com uma alta produção de IFN-&#947; e desvio da resposta imune para o tipo Th1, a proteção contra helmintos está associada a uma resposta Th2. Devido a este viés da resposta imune, indivíduos infectados pelo HTLV-1 apresentam redução na produção de IL-4, IL-5, IL-13 e IgE, componentes participantes dos mecanismos de defesa contra S. stercoralis. Estas anormalidades constituem a base para a ocorrência de maior freqüência e de formas mais graves da estrongiloidíase em pacientes infectados pelo HTLV-1.

Desafios na era pós genômica para o desenvolvimento de testes laboratoriais para o diagnóstico da hanseníase

O diagnóstico da hanseníase se baseia em manifestações clínicas e não existe teste laboratorial para diagnosticar casos assintomáticos ou para prever progressão da doença entre indivíduos expostos. Novas análises genômicas comparativas in silico e ferramentas de biologia molecular têm sido empregadas para revelar proteínas exclusivas do Mycobacterium leprae que apresentem potencial aplicação diagnóstica. A hanseníase tuberculóide paucibacilar (PB) apresenta baixo nível de anticorpos e forte resposta imune celular (RIC) tipo Th1/interferon gamma (IFN-&#947;). A doença lepromatosa multibacilar (MB) apresenta sorologia positiva e fraca RIC. Portanto, testes laboratoriais para diagnosticar hanseníase PB e MB devem contemplar testes de RIC e sorologia. Proteínas recombinantes do Mycobacterium leprae sorologicamente reativas podem ser incorporadas ao antígeno PGLI para melhorar o diagnóstico sorológico de pacientes MB. Proteínas recombinantes e peptídeos sintéticos do Mycobacterium leprae têm sido testados em ensaios de RIC/IFN-&#947; para diagnosticar casos PB. Sorologia anti-PGLI modificada incorporando novos antígenos do Mycobacterium leprae e ensaios baseados na RIC/produção de IFN-&#947; devem permitir a detecção precoce de casos MB e PB em países endêmicos.

Cytokine expression in the duodenal mucosa of patients with visceral leishmaniasis

INTRODUCTION: Visceral leishmaniasis (VL) is a neglected tropical disease with a complex immune response in different organs. This pattern of organ-specific immune response has never been evaluated in the gastrointestinal tract. The aim of this study was to determine the in situ immune response in duodenal biopsies on patients with VL. METHODS: A case-control study was conducted on 13 patients with VL in comparison with nine controls. The immune response was evaluated using immunohistochemistry, for CD4, CD8, CD68, IL-4, IFN-&#947;, TNF-α and IL-10. Histological findings from the villi, crypts and inflammatory process were analyzed. RESULTS: All the cases of VL presented Leishmania antigens. No antigen was detected in the control group. The villus size was greater in the VL patients (p < 0.05). CD68 (macrophages) and CD4 levels were higher in the VL patients (p < 0.05). No differences in the expression of CD8, TNF-α, IL-10 or IL-4 were demonstrated. The number of cells expressing IFN-&#947; was lower in the VL patients (p < 0.05). CONCLUSIONS: Low levels of cytokines were found in the gastrointestinal tract of patients with VL. This pattern was not found in other organs affected by the disease. Immunotolerance of this tissue against Leishmania could explain these findings, as occurs with intestinal bacteria.

Atypical lymphocytosis in leptospirosis: a cohort of hospitalized cases between 1996 and 2009 in State of Rio de Janeiro, Brazil

INTRODUCTION: Leptospirosis is a zoonotic disease found in tropical and temperate countries, and its clinical diagnostic confusion with arboviruses (dengue fever, oropouche fever and yellow fever), Brazilian spotted fever, viral hepatitis and hantaviruses has been an ongoing public health concern. The aim of this observational study was to demonstrate an association between findings of atypical lymphocytosis and the progression of endemic leptospirosis. METHODS: A retrospective analysis was performed on the demographic, epidemiological, clinical and laboratory aspects of 27 human leptospirosis cases that occurred over a period of 13 years (1996-2009) with no reported epidemic outbreaks in Rio de Janeiro, Brazil. RESULTS: The overall mortality rate was 11.1% in our cohort of hospitalized cases. However, there was no mortality among patients with atypical lymphocytosis (OR = 11.1; 95% CI = 1.12-110.9; p = 0.04). Two patients who were in the septicemic phase showed signs of expansion of &#947;δ T cell responses in peripheral blood. CONCLUSIONS: Atypical lymphocytosis may be observed in patients with leptospirosis. Our observations suggest that these atypical leukocyte subsets are associated with partial protection during the disease course of leptospirosis.

Oral coinfection can stress peripheral lymphocyte to inflammatory activity in leprosy

INTRODUCTION: This study evaluated the intracellular profile of interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10) and interferon-&#947; (IFN-&#947;) in peripheral blood mononuclear cells (PBMCs) from leprosy patients based on oral infections presence to determine whether these coinfections could be associated with pro-inflammatory activity in leprosy. METHODS: Leprosy patients regardless of clinical form and specific leprosy treatment (n=38) were divided into two groups: Group I - leprosy patients with oral infections (n=19), and Group II - leprosy patients without oral infections (n=19). Non-leprosy patients presenting oral infections were assigned to the control Group (n=10). Intracellular IL-2, IL-4, IL-10 and IFN-&#947; production was evaluated by flow cytometry (FACS) before and 7 days after controlling the oral infection in the Group I, before and 7 days after dental prophylaxis in the Group II, and during oral infection process in control Group. RESULTS: Low percentages of CD3+ lymphocytes bearing IL-2, IL-10 and IFN-&#947; were observed in the Group I and Group II at baseline and 7 days after therapy or prophylaxis compared to controls. Group I showed reduced percentages of IL-4 at baseline and 7 days after therapy compared to controls, or at baseline of Group II, and the Group II showed reduced percentages of CD3+ cells bearing IL-4 compared to control. An increase of the percentages of CD3+cells bearing IL-4 was observed in the Group I after the oral infections treatment. CONCLUSIONS: The occurrence of oral infections favors the intracellular cytokines expression and, probably, the inflammatory reaction operating as a stimulatory signal triggering the leprosy reactions.

Involvement and interactions of different immune cells and their cytokines in human visceral leishmaniasis

Visceral leishmaniasis (VL) or kala-azar, a disseminated infection of the lymphoreticular system of the body, is marked by severe defect in immune system of the host. Successful cure of VL depends on the immune status of the host in combination with the effects of the antileishmanial drugs. The rationale approach towards eradication of this disease would be to potentiate the immune functioning of the host in addition to parasite killing. This review deals with different aspects of adaptive and innate immune responses and explores their role in protection or pathogenesis of VL. IL-10 has emerged as the principal cytokine responsible for disease pathogenesis, although evidences regarding its source during active VL remain inconclusive. On the other hand, IFN&#947;, under the influence of IL-12, is mostly correlated with healing of the disease. Chemokines are important in mounting cell-mediated immune response as they can prevent parasite invasion in association with cytokines. Different types of T cells like CD4, CD8 and NK T cells also contribute to the immunology of this disease. In spite of conflicting reports, the role of regulatory T cells in VL pathogenesis is important. Recently discovered Th17 subset and its different members have been reported to perform diverse functions in the course of VL and leishmaniasis as a whole. Innate immune responses, depending on the cell types, are essential in early parasite detection and subsequent development of an efficient NK cell response. Immunotherapy targeting IL-10 could be looked upon as an interesting option for the treatment of VL.

Severe visceral leishmaniasis in children: the relationship between cytokine patterns and clinical features

Introduction The relationship between severe clinical manifestations of visceral leishmaniasis (VL) and immune response profiles has not yet been clarified, despite numerous studies on the subject. This study aimed to investigate the relationship between cytokine profiles and the presence of immunological markers associated with clinical manifestations and, particularly, signs of severity, as defined in a protocol drafted by the Ministry of Health (Brazil). Methods We conducted a prospective, descriptive study between May 2008 and December 2009. This study was based on an assessment of all pediatric patients with VL who were observed in a reference hospital in Maranhão. Results Among 27 children, 55.5% presented with more than one sign of severity or warning sign. Patients without signs of severity or warning signs and patients with only one warning sign had the highest interferon-gamma (IFN-&#947;) levels, although their interleukin 10 (IL-10) levels were also elevated. In contrast, patients with the features of severe disease had the lowest IFN-&#947; levels. Three patients who presented with more than two signs of severe disease died; these patients had undetectable interleukin 2 (IL-2) and IFN-&#947; levels and low IL-10 levels, which varied between 0 and 36.8pg/mL. Conclusions Our results showed that disease severity was associated with low IFN-&#947; levels and elevated IL-10 levels. However, further studies with larger samples are needed to better characterize the relationship between disease severity and cytokine levels, with the aim of identifying immunological markers of active-disease severity.

Comparative study of lymphocytes from individuals that were vaccinated and unvaccinated against the pandemic 2009-2011 H1N1 influenza virus in Southern Brazil

ABSTRACTINTRODUCTION:While no single factor is sufficient to guarantee the success of influenza vaccine programs, knowledge of the levels of immunity in local populations is critical. Here, we analyzed influenza immunity in a population from Southern Brazil, a region with weather conditions that are distinct from those in the rest of country, where influenza infections are endemic, and where greater than 50% of the population is vaccinated annually.METHODS:Peripheral blood mononuclear cells were isolated from 40 individuals. Of these, 20 had received the H1N1 vaccine, while the remaining 20 were unvaccinated against the disease. Cells were stimulated in vitro with the trivalent post-pandemic influenza vaccine or with conserved major histocompatibility complex I (MHC I) peptides derived from hemagglutinin and neuraminidase. Cell viability was then analyzed by [3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide)]-based colorimetric assay (MTT), and culture supernatants were assayed for helper T type 1 (Th1) and Th2-specific cytokine levels.RESULTS:Peripheral blood lymphocytes from vaccinated, but not unvaccinated, individuals exhibited significant proliferation in vitro in the presence of a cognate influenza antigen. After culturing with vaccine antigens, cells from vaccinated individuals produced similar levels of interleukin (IL)-10 and interferon (IFN)-&#947;, while those from unvaccinated individuals produced higher levels of IFN-&#947; than of IL-10.CONCLUSIONS:Our data indicate that peripheral blood lymphocytes from vaccinated individuals are stimulated upon encountering a cognate antigen, but did not support the hypothesis that cross-reactive responses related to previous infections can ameliorate the immune response. Moreover, monitoring IL-10 production in vaccinated individuals could comprise a valuable tool for predicting disease evolution.

Sodium nitroprusside has leishmanicidal activity independent of iNOS

Abstract: INTRODUCTION: Leishmaniasis is a zoonotic disease caused by protozoa of the genus Leishmania . Cutaneous leishmaniasis is the most common form, with millions of new cases worldwide each year. Treatments are ineffective due to the toxicity of existing drugs and the resistance acquired by certain strains of the parasite. METHODS: We evaluated the activity of sodium nitroprusside in macrophages infected with Leishmania (Leishmania) amazonensis . Phagocytic and microbicidal activity were evaluated by phagocytosis assay and promastigote recovery, respectively, while cytokine production and nitrite levels were determined by ELISA and by the Griess method. Levels of iNOS and 3-nitrotyrosine were measured by immunocytochemistry. RESULTS: Sodium nitroprusside exhibited in vitro antileishmanial activity at both concentrations tested, reducing the number of amastigotes and recovered promastigotes in macrophages infected with L. amazonensis . At 1.5µg/mL, sodium nitroprusside stimulated levels of TNF-α and nitric oxide, but not IFN-&#947;. The compound also increased levels of 3-nitrotyrosine, but not expression of iNOS, suggesting that the drug acts as an exogenous source of nitric oxide. CONCLUSIONS: Sodium nitroprusside enhances microbicidal activity in Leishmania -infected macrophages by boosting nitric oxide and 3-nitrotyrosine.

Os Óleos Essenciais de Piper reticulatum L. e P. crassinervium H. B. K.

Os óleos essenciais das folhas e galhos finos de Piper reticulatum e de P. crassinervium, coletados na região norte do Brasil, foram obtidos por arraste à vapor e analisados através de GC/MS. O óleo de P. reticulatum é constituido principalmente por β-elemeno (24,6%) e β-cariofileno (16,7%). Os principais compostos identificados no óleo de P. crassinervium foramβ-cariofileno (17,7%), &#947;-elemeno (14,4%) e β-elemeno (10,9%).

Tocopherol profile of Brazil nut oil from different geographic areas of the Amazon region

The tocopherol content of Brazil nut oil from different Amazon regions (Manicoré-AM, Rio Preto da Eva-AM, São João da Baliza-RR, Caroebe-RR, Belém-PA, and Xapurí-AC) was investigated by normal-phase high-performance liquid chromatography. For all authentic oils, two isomers: α- and &#947;-tocopherols were observed (37.92-74.48 µg g-1, 106.88-171.80 µg g-1, respectively), and their levels were relatively constant among the oils having these geographic origins, which would enable to distinguish Brazil nut oil from other plant oils for authentication purposes. Commercial Brazil nut oils were also evaluated, and some of these oils demonstrated a tocopherol content that was very different from that of the authentic oils. Therefore, we suggest that the tocopherol profile of Brazil nut oil can be useful chemical marker for quality control and authentication.

Validade discriminante do Teste de Aprendizagem Auditivo-Verbal de Rey: comparação entre idosos normais e idosos na fase inicial da doença de Alzheimer

Objetivo: Este estudo transversal visa avaliar a validade discriminante do Teste de Aprendizagem Auditivo-Verbal de Rey (RAVLT), ao comparar uma amostra de idosos normais com uma de pacientes na fase inicial da doença de Alzheimer (DA). Métodos: Pacientes na fase inicial da DA (n = 35) e controles saudáveis (n = 35) pareados de acordo com a idade e a escolaridade foram submetidos ao Teste de Aprendizagem Auditivo-Verbal de Rey, ao Miniexame do Estado Mental e à Escala de Depressão Geriátrica. O desempenho dos dois grupos foi comparado por meio do teste de Mann-Whitney em cada etapa do RAVLT e, mediante a análise ROC, foi avaliada a validade discriminante do teste nas duas populações estudadas. Resultados: O grupo na fase inicial da DA teve desempenho significativamente pior em todas as etapas do RAVLT quando comparado ao grupo controle, e as etapas demonstraram bom poder diagnóstico, com áreas sobre a curva ROC oscilando entre 0,806 e 0,989 (A1 = 0,806; A2 = 0,869; A3 = 0,958; A4 = 0,947; A5 = 0,989; A6 = 0,962; A7 = 0,985; TOTAL = 0,975; LOT = 0,895; REC = 0,915). Conclusão: Os resultados sugerem que o Teste de Aprendizagem Auditivo-Verbal de Rey é eficaz para discriminar idosos normais de idosos na fase inicial da doença de Alzheimer.

Estudo da distribuição dos pontos de máximo ou de mínimo de equações de regressão de segundo grau

Sabe-se que é comum usar-se a regressão quadrática (Y = a.r² + bX -f- c) para determinar a dose econômica de adubação. O ponto de máximo ou de mínimo será X = - - b/2c , onde b e c possuem distribuição normal. X Teste trabalho cogita-se de estudar a distribuição gerada pelo quociente de duas variáveis pertencentes a uma distribuição normal. Calcularam-se as estatísticas &#947;1 e &#947;2 de Fisher, e a elas se aplicou a prova de t. Também se obtiveram os momentos 3.° e 4.°. Os resultados obtidos mostram que na maioria dos casos a distribuição de X se afasta muito da normal.

Flutuação populacional circanual de coleópteros em granja avícola, em Pelotas, RS, Brasil

Com o objetivo de conhecer a flutuação populacional de coleópteros na granja de aves do Conjunto Agrotécnico Visconde da Graça, em Pelotas, RS, foram utilizados seis métodos de coleta: 1 (fezes de galinhas com 0 a 7 dias); 2 (7 a 14 dias); 3 (14 a 21 dias); 4 (0 a 21 dias); 5 (fezes acumuladas) e 6 (armadilhas de tubo). Análises de regressão polinomial foram realizadas independentemente dos métodos de coleta. O estudo foi realizado de agosto de 1998 a julho de 1999. Foram capturados 12.449 coleópteros representados pelas seguintes espécies Carcinops troglodytes (Paykull, 1811) (Histeridae) (6.444); Alphitobius diaperinus (Panzer, 1797) (Tenebrionidae) (2.896); Somotrichus unifasciatus (Dejean, 1792) (Carabidae) (1.190); Gnathocerus cornutus (Fabricius, 1798) (Tenebrionidae) (947); Euxestus sp. (Cerylonidae) (394); Euspilotus rubriculus (Marseul, 1855) (Histeridae) (213); Ontholestes sp. (Staphylinidae) (190); Dactylosternum sp. (Hydrophilidae) (93); Mezium americanum (Laporte, 1840) (Ptinidae) (43); Palorus subdepressus (Wollaston, 1864) (Tenebrionidae) (27); Tenebroides mauritanicus (L., 1758) (Trogossitidae) (8); Xyleborus ferrugineus (Fabricius, 1801) (Scolytidae) (1); e espécimens não identificados de Dermestidae (3). O maior número de espécies ocorreu nos meses de março e julho e o menor, no mês de setembro. Em março foi registrada a maior abundância de coleópteros (2.159), enquanto que o menor índice de captura foi assinalado em outubro (633). A flutuação populacional foi estimada para C. troglodytes, A. diaperinus, S. unifasciatus, G. cornutus, Euxestus sp., E. rubriculus, Ontholestes sp. e Dactylosternum sp.