Assessment of PCR in the detection of Leishmania spp in experimentally infected individual phlebotomine sandflies (Diptera: Psychodidae: Phlebotominae)

DNA amplification by the polymerase chain reaction (PCR) was applied in the investigation of the presence of Leishmania (Kinetoplastida: Trypanosomatidae) parasites in single phlebotomine sandflies. Three phlebotomine/parasite pairs were used: Lutzomyia longipalpis/Leishmania chagasi, Lutzomyia migonei/Leishmania amazonensis and Lutzomyia migonei/Leishmania braziliensis, all of them incriminated in the transmission of visceral or cutaneous leishmaniasis. DNA extraction was performed with whole insects, with no need of previous digestive tract dissection or pooling specimens. The presence of either mouse blood in the digestive tract of the sandflies or the digestive tract itself did not interfere in the PCR. Infection by as few as 10 Leishmania sp. per individual were sufficient for DNA amplification with genus-specific primers. Using primers for L. braziliensis and L. mexicana complexes, respectively, it was possible to discriminate between L. braziliensis and L. amazonensis in experimentally infected vectors (L. migonei).

Specific treatment of hepatosplenic schistosomiasis can increase T-lymphocyte reactivity

It has been recognized that Schistosoma mansoni infection causes depression of T-cell responsiveness. In this study we have evaluated whether immunodepression associated to schistosomiasis could be reverted by specific treatment. T-cell immune response was assessed by means of intradermal tests using recall antigens in a group of 22 patients with hepatosplenic schistosomiasis, one year after treatment with oxamniquine and compared with a group of untreated hepatosplenic patients. Only 27% of treated patients presented complete anergy to all tested antigens, in marked contrast to 80% unresponsiveness showed by hepatosplenic patients without treatment. Although most of the treated individuals showed some response to the tested antigens, in some individuals this unresponsiveness still persisted after treatment. Anergy was not found in any normal individual of the control group. It was concluded that Schistosoma mansoni infected patients may recover their normal immune responsiveness after the elimination of the worm by treatment

Antigens of worms and eggs showed a differentiated detection of specific IgG according to the time of Schistosoma mansoni infection in mice

INTRODUCTION: The correlation between the immunological assay and the antibody titer can offer a tool for the experimental analysis of different phases of the disease. METHODS: Two simple immunological assays for Schistosoma mansoni in mice sera samples based on specific IgG detection for worms soluble antigens and eggs soluble antigens were standardized and evaluated in our laboratory. Fifty mice were used in negative and positive groups and the results obtained by enzyme-linked immunosorbent assays (ELISA) assays were compared with the number of worms counted and the IgG titers at different times of infection. RESULTS: Data showed that ELISA using adult worm antigens (ELISA-SWAP) presented a satisfactory correlation between the absorbance value of IgG titers and the individual number of worms counted after perfusion technique (R²=0.62). In addition, ELISA-SWAP differentially detected positive samples with 30 and 60 days post infection (p=0.011 and 0.003, respectively), whereas ELISA using egg antigens (ELISA-SEA) detected samples after 140 days (p=0.03). CONCLUSIONS: These data show that the use of different antigens in immunological methods can be used as potential tools for the analysis of the chronological evolution of S. mansoni infection in murine schistosomiasis. Correlations with human schistosomiasis are discussed.

Sex-Specific Equations to Estimate Maximum Oxygen Uptake in Cycle Ergometry

AbstractBackground:Aerobic fitness, assessed by measuring VO2max in maximum cardiopulmonary exercise testing (CPX) or by estimating VO2max through the use of equations in exercise testing, is a predictor of mortality. However, the error resulting from this estimate in a given individual can be high, affecting clinical decisions.Objective:To determine the error of estimate of VO2max in cycle ergometry in a population attending clinical exercise testing laboratories, and to propose sex-specific equations to minimize that error.Methods:This study assessed 1715 adults (18 to 91 years, 68% men) undertaking maximum CPX in a lower limbs cycle ergometer (LLCE) with ramp protocol. The percentage error (E%) between measured VO2max and that estimated from the modified ACSM equation (Lang et al. MSSE, 1992) was calculated. Then, estimation equations were developed: 1) for all the population tested (C-GENERAL); and 2) separately by sex (C-MEN and C-WOMEN).Results:Measured VO2max was higher in men than in WOMEN: -29.4 ± 10.5 and 24.2 ± 9.2 mL.(kg.min)-1 (p < 0.01). The equations for estimating VO2max [in mL.(kg.min)-1] were: C-GENERAL = [final workload (W)/body weight (kg)] x 10.483 + 7; C-MEN = [final workload (W)/body weight (kg)] x 10.791 + 7; and C-WOMEN = [final workload (W)/body weight (kg)] x 9.820 + 7. The E% for MEN was: -3.4 ± 13.4% (modified ACSM); 1.2 ± 13.2% (C-GENERAL); and -0.9 ± 13.4% (C-MEN) (p < 0.01). For WOMEN: -14.7 ± 17.4% (modified ACSM); -6.3 ± 16.5% (C-GENERAL); and -1.7 ± 16.2% (C-WOMEN) (p < 0.01).Conclusion:The error of estimate of VO2max by use of sex-specific equations was reduced, but not eliminated, in exercise tests on LLCE.

Schistosoma mansoni in a low-prevalence area in Brazil: the importance of additional methods for the diagnosis of hard-to-detect individual carriers by low-cost immunological assays

Schistosomiasis diagnosis is based on the detection of eggs in the faeces, which is laborious and lacks sensitivity, especially for patients with a low parasite burden. Immunological assays for specific antibody detection are available, but they usually demonstrate low sensitivity and/or specificity. In this study, two simple immunological assays were evaluated for the detection of soluble Schistosoma mansoni adult worm preparation (SWAP) and egg-specific IgGs. These studies have not yet been evaluated for patients with low parasite burdens. Residents of an endemic area in Brazil donated sera and faecal samples for our study. The patients were initially diagnosed by a rigorous Kato-Katz analysis of 18 thick smears from four different stool samples. The ELISA-SWAP was successful for human diagnosis with 90% sensitivity and specificity, confirming the Kato-Katz diagnosis with nearly perfect agreement, as seen by the Kappa index (0.85). Although the ELISA-soluble S. mansoni egg antigen was 85% sensitive, it exhibited low specificity (80%; Kappa index: 0.75) and was more susceptible to cross-reactivity. We believe that immunological assays should be used in conjunction with Kato-Katz analysis as a supplementary tool for the diagnosis of schistosomiasis for patients with low infection burdens, which are usually hard to detect.

Mycobacterium tuberculosis epitope-specific interferon-g production in healthy Brazilians reactive and non-reactive to tuberculin skin test

The interferon (IFN)-γ response to peptides can be a useful diagnostic marker of Mycobacterium tuberculosis (MTB) latent infection. We identified promiscuous and potentially protective CD4+ T-cell epitopes from the most conserved regions of MTB antigenic proteins by scanning the MTB antigenic proteins GroEL2, phosphate-binding protein 1 precursor and 19 kDa antigen with the TEPITOPE algorithm. Seven peptide sequences predicted to bind to multiple human leukocyte antigen (HLA)-DR molecules were synthesised and tested with IFN-γ enzyme-linked immunospot (ELISPOT) assays using peripheral blood mononuclear cells (PBMCs) from 16 Mantoux tuberculin skin test (TST)-positive and 16 TST-negative healthy donors. Eighty-eight percent of TST-positive donors responded to at least one of the peptides, compared to 25% of TST-negative donors. Each individual peptide induced IFN-γ production by PBMCs from at least 31% of the TST-positive donors. The magnitude of the response against all peptides was 182 ± 230 x 106 IFN-γ spot forming cells (SFC) among TST-positive donors and 36 ± 62 x 106 SFC among TST-negative donors (p = 0.007). The response to GroEL2 (463-477) was only observed in the TST-positive group. This combination of novel MTB CD4 T-cell epitopes should be tested in a larger cohort of individuals with latent tuberculosis (TB) to evaluate its potential to diagnose latent TB and it may be included in ELISPOT-based IFN-γ assays to identify individuals with this condition.

DNA encoding individual mycobacterial antigens protects mice against tuberculosis

Over the last few years, some of our experiments in which mycobacterial antigens were presented to the immune system as if they were viral antigens have had a significant impact on our understanding of protective immunity against tuberculosis. They have also markedly enhanced the prospects for new vaccines. We now know that individual mycobacterial protein antigens can confer protection equal to that from live BCG vaccine in mice. A critical determinant of the outcome of immunization appears to be the degree to which antigen-specific cytotoxic T cells are generated by the immune response. Our most recent studies indicate that DNA vaccination is an effective way to establish long-lasting cytotoxic T cell memory and protection against tuberculosis.

Avaliação dos limiares auditivos com e sem equipamento de proteção individual

Os autores realizaram estudo caso-controle audiométrico em indivíduos com e sem protetor auricular auditivo. OBJETIVOS: O objetivo do estudo foi avaliar a real atenuação individual dado pelos protetores. MATERIAL E MÉTODO: Foram avaliados 30 indivíduos (ou 60 orelhas) de diferentes atividades profissionais, de ambos os sexos, com idades entre 20 e 58 anos, apresentando audição normal e tendo realizado repouso auditivo de 10 horas, submetidos a exame audiométrico com e sem protetor auricular auditivo, no período de fevereiro a julho de 2003, utilizando protetor tipo plugue. Avaliou-se as audiometrias nas vias aérea e óssea em freqüências de 500 a 4000Hz. RESULTADOS: Os resultados foram analisados estatisticamente e comparados aos dados fornecidos pelo fabricante. Assim se observou em ouvido real os níveis de atenuação auditiva obtidos com o uso destes produtos. CONCLUSÃO: Os resultados permitiram chegar à conclusão de que os índices fornecidos pelos fabricantes foram compatíveis com os que obtive nos testes.

Efeitos de treinamento nos desempenhos individual e organizacional

O objetivo deste estudo foi desenvolver um método para identificar efeitos de treinamento nos desempenhos dos indivíduos e da organização, combinação ainda rara na literatura científica. Foi realizado no Banco do Brasil, com 218 profissionais que fizeram um MBA em Marketing. Os dados foram coletados por meio de entrevistas e questionários de auto e heteroavaliação. Nos questionários foi avaliada a contribuição do treinamento para a melhoria do desempenho individual e organizacional, e o suporte à transferência de treinamento. Nas entrevistas foram levantados indicadores de melhoria do desempenho organizacional para os maiores impactos percebidos pelos respondentes. Os resultados indicam que, para a maioria dos desempenhos individuais e organizacionais, o impacto percebido do treinamento foi avaliado entre bom e ótimo. A metodologia utilizada foi considerada adequada para a identificação de melhorias nos desempenhos organizacionais, percebidos como os mais afetados pelo treinamento. Também foram discutidas implicações metodológicas, teóricas e perspectivas futuras.

Fatores críticos no comportamento do gestor público responsável por compras sustentáveis: diferenças entre consumo individual e organizacional

O presente artigo investiga o comportamento de gestores públicos, identificando os fatores críticos que determinam o processo decisório de compra sustentável, seja no contexto de compras individuais (ou domésticas) ou no contexto de compras organizacionais (nesse caso, refere-se às compras públicas sustentáveis, conceito recentemente inserido na legislação brasileira). Em seguida à aplicação de questionários a um grupo de gestores públicos encarregados da elaboração de editais e condução de certames licitatórios, foram realizadas inferências estatísticas que determinaram as similitudes entre os dois contextos. A partir dos resultados, observou-se diferenças relacionadas com o contexto de compra envolvido, em especial naquelas que envolvem preço, determinações da legislação e tempo gasto nas especificações dos produtos a serem adquiridos. Chamam atenção, ademais, o fato de que alguns padrões pró-ambientais se mostram bem estabelecidos, evidenciado o resultado de campanhas desenvolvidas no âmbito da instituição.

Esterilidad individual en Rhodnius prolixus Stal. tratados con metepa (oxido de tris (1-(2 metil) aziridinil) fosfuro), y accion del mismo sobre Trypanosoma cruzi

Se analizan los factores que interfieren en la esterilidad inducida químicamente en R. Prolixus mediante aplicaciones tópicas en distintas dosis de metepa. Se sugiere profundizar el estudio de la susceptibilidad individual para conocer la composición de la población frente al quimioesterilizante y la probable aparición de resistencia. Se investiga además la acción del metepa sobre T. cruzi en triatominos infectados experimentalmente, obteniéndose resultados negativos.

Antinomias e "suturas" epistemológicas entre biológico-social e individual-coletivo no âmbito da epidemiologia social

A complexidade do objeto epidemiológico tem suscitado, ao longo do tempo, discussão sobre alguns elementos que o compõem, assumindo, muitas vezes, a forma de antinomias. Utilizando-se como substrato textos fundamentais, analisaram-se, no âmbito da epidemiologia social, a formulação e a proposta de solução para as antinomias biológico-social e individual-coletivo. Criticou-se a validade teórica das saídas apontadas pelo discurso epidemiológico-social para resolver o "conflito de leis" que permeia o referido objeto. Destacou-se o conceito marxista-helleriano de indivíduo para contribuir com a "sutura" dos três planos da realidade: o universal, o particular e o singular.

Human cryptosporidiosis: detection of specific antibodies in the serum by an indirect immunofluorescence

Cryptosporidium sp., a coccidian parasite usually found in the faeces of cattle, has been recently implicated as an agent of human intestinal disease, mainly in immunocompromised patients. In the study realized, by an indirect immunofluorescence technique, specific immunoglobulins (IgG and IgM) have been demonstrated in human serum against Cryptosporidium oocysts. Purified oocysts were used as antigens in the indirect immunofluorecence assay. After analyzing this test in sera from selected groups of patients, the frequency of both specific IgG and IgM of immunocompetent children who were excreting oocysts in their faeces was 62% and in children with negative excretion of oocysts was 20% and 40%, respectively. In adults infected with the human immunodeficiency virus (HIV) and who were excreting Cryptosporidium in their stools, the frequency was 57% for IgG but only 2% for IgM. Twenty three percent of immunocompromised adults with not determined excretion of oocysts in their stools had anti-Cryptosporidium IgG in their sera. Children infected with human immunodeficiency virus had no IgM and only 14% had IgG detectable in their sera. The indirect immunoflorescence assay, when used with other parasitological techniques appears to be useful for retrospective population studies and for diagnosis of acute infection. The humoral immune response of HIV positive patients to this protozoan agent needs clarification.

Multidimensional aspects related to shiftworkers' health and well-being

The impact of shift and night work on health shows a high inter- and intra-individual variability, both in terms of kind of troubles and temporal occurrence, related to various intervening factors dealing with individual characteristics, lifestyles, work demands, company organisation, family relations and social conditions. The way we define "health" and "well-being" can significantly influence appraisals, outcomes and interventions. As the goal is the optimisation of shiftworkers' health, it is necessary to go beyond the health protection and to act for health promotion. In this perspective, not only people related to medical sciences, but many other actors (ergonomists, psychologists, sociologists, educators, legislators), as well as shiftworkers themselves. Many models have been proposed aimed at describing the intervening variables mediating and/or moderating the effects; they try to define the interactions and the pathways connecting risk factors and outcomes through several human dimensions, which refer to physiology, psychology, pathology, sociology, ergonomics, economics, politics, and ethics. So, different criteria can be used to evaluate shiftworkers' health and well-being, starting from biological rhythms and ending in severe health disorders, passing through psychological strain, job dissatisfaction, family perturbation and social dis-adaptation, both in the short- and long-term. Consequently, it appears rather arbitrary to focus the problem of shiftworkers' health and tolerance only on specific aspects (e.g. individual characteristics), but a systemic approach appears more appropriate, able to match as many variables as possible, and aimed at defining which factors are the most relevant for those specific work and social conditions. This can support a more effective and profitable (for individuals, companies, and society) adoption of preventive and compensative measures, that must refer more to "countervalues" rather than to "counterweights".