Differential expression of the costimulatory molecules CD86, CD28, CD152 and PD-1 correlates with the host-parasite outcome in leprosy

Leprosy is a spectral disease exhibiting two polar sides, namely, lepromatous leprosy (LL) characterised by impaired T-cell responses and tuberculoid leprosy in which T-cell responses are strong. Proper T-cell activation requires signalling through costimulatory molecules expressed by antigen presenting cells and their ligands on T-cells. We studied the influence of costimulatory molecules on the immune responses of subjects along the leprosy spectrum. The expression of the costimulatory molecules was evaluated in in vitro-stimulated peripheral blood mononuclear cells of lepromatous and tuberculoid patients and healthy exposed individuals (contacts). We show that LL patients have defective monocyte CD86 expression, which likely contributes to the impairment of the antigen presentation process and to patients anergy. Accordingly, CD86 but not CD80 blockade inhibited the lymphoproliferative response to Mycobacterium leprae. Consistent with the LL anergy, there was reduced expression of the positive signalling costimulatory molecules CD28 and CD86 on the T-cells in these patients. In contrast, tuberculoid leprosy patients displayed increased expression of the negative signalling molecules CD152 and programmed death-1 (PD-1), which represents a probable means of modulating an exacerbated immune response and avoiding immunopathology. Notably, the contacts exhibited proper CD86 and CD28 expression but not exacerbated CD152 or PD-1 expression, suggesting that they tend to develop a balanced immunity without requiring immunosuppressive costimulatory signalling.

Cross-reactive anti-PfCLAG9 antibodies in the sera of asymptomatic parasite carriers of Plasmodium vivax

The PfCLAG9 has been extensively studied because their immunogenicity. Thereby, the gene product is important for therapeutics interventions and a potential vaccine candidate. Antibodies against synthetic peptides corresponding to selected sequences of the Plasmodium falciparum antigen PfCLAG9 were found in sera of falciparum malaria patients from Rondônia, in the Brazilian Amazon. Much higher antibody titres were found in semi-immune and immune asymptomatic parasite carriers than in subjects suffering clinical infections, corroborating original findings in Papua Guinea. However, sera of Plasmodium vivax patients from the same Amazon area, in particular from asymptomatic vivax parasite carriers, reacted strongly with the same peptides. Bioinformatic analyses revealed regions of similarity between P. falciparum Pfclag9 and the P. vivax ortholog Pvclag7. Indirect fluorescent microscopy analysis showed that antibodies against PfCLAG9 peptides elicited in BALB/c mice react with human red blood cells (RBCs) infected with both P. falciparum and P. vivax parasites. The patterns of reactivity on the surface of the parasitised RBCs are very similar. The present observations support previous findings that PfCLAG9 may be a target of protective immune responses and raises the possibility that the cross reactive antibodies to PvCLAG7 in mixed infections play a role in regulate the fate of Plasmodium mixed infections.

Ultrastructural description of Ceratomyxa microlepis sp. nov. (Phylum Myxozoa): a parasite infecting the gall bladder of Hemiodus microlepis, a freshwater teleost from the Amazon River

A new ceratomyxid parasite was examined for taxonomic identification, upon being found infecting the gall bladder of Hemiodus microlepis (Teleostei: Hemiodontidae), a freshwater teleost collected from the Amazon River, Brazil. Light and transmission electron microscopy revealed elongated crescent-shaped spores constituted by two asymmetrical shell valves united along a straight sutural line, each possessing a lateral projection. The spores body measured 5.2 ± 0.4 µm (n = 25) in length and 35.5 ± 0.9 µm (n = 25) in total thickness. The lateral projections were asymmetric, one measuring 18.1 ± 0.5 µm (n = 25) in thickness and the other measuring 17.5 ± 0.5 µm (n = 25) in thickness. Two equal-sized subspherical polar capsules measuring 2.2 ± 0.3 µm in diameter were located at the same level, each possessing a polar filament with 5-6 coils. The sporoplasm was binucleate. Considering the morphometric data analyzed from the microscopic observations, as well as the host species and its geographical location, this paper describes a new myxosporean species, herein named Ceratomyxa microlepis sp. nov.; therefore representing the first description of a freshwater ceratomyxid from the South American region.

Submicroscopic malaria parasite carriage: how reproducible are polymerase chain reaction-based methods?

The polymerase chain reaction (PCR)-based methods for the diagnosis of malaria infection are expected to accurately identify submicroscopic parasite carriers. Although a significant number of PCR protocols have been described, few studies have addressed the performance of PCR amplification in cases of field samples with submicroscopic malaria infection. Here, the reproducibility of two well-established PCR protocols (nested-PCR and real-time PCR for the Plasmodium 18 small subunit rRNA gene) were evaluated in a panel of 34 blood field samples from individuals that are potential reservoirs of malaria infection, but were negative for malaria by optical microscopy. Regardless of the PCR protocol, a large variation between the PCR replicates was observed, leading to alternating positive and negative results in 38% (13 out of 34) of the samples. These findings were quite different from those obtained from the microscopy-positive patients or the unexposed individuals; the diagnosis of these individuals could be confirmed based on the high reproducibility and specificity of the PCR-based protocols. The limitation of PCR amplification was restricted to the field samples with very low levels of parasitaemia because titrations of the DNA templates were able to detect < 3 parasites/µL in the blood. In conclusion, conventional PCR protocols require careful interpretation in cases of submicroscopic malaria infection, as inconsistent and false-negative results can occur.

New records of Mansonella ozzardi: a parasite that is spreading from the state of Amazonas to previously uninfected areas of the state of Acre in the Purus River region

Mansonella ozzardi infections are common in the riverside communities along the Solimões, Negro and Purus Rivers in the state of Amazonas (AM). However, little is known about the presence of this parasite in communities located in regions bordering AM and the state of Acre. The prevalence rate of M. ozzardi infections was determined in blood samples from volunteers according to the Knott method. A total of 355 volunteers from six riverine communities were enrolled in the study and 65 (18.3%) were found to be infected with M. ozzardi. As expected, most of the infections (25%) occurred in individuals involved in agriculture, cattle rearing and fishing and an age/sex group analysis revealed that the prevalence increased beginning in the 40-50-years-of-age group and reached 33% in both sexes in individuals over 50 years of age. Based on the described symptomatology, articular pain and headache were found to be significantly higher among infected individuals (56 and 65% prevalence, respectively, p < 0.05). Sera from volunteers were subjected to ELISA using a cocktail of recombinant proteins from Onchocerca volvulus to evaluate the specificity of the test in an endemic M. ozzardi region. No cross-reactions between M. ozzardi-infected individuals and recombinant O. volvulusproteins were detected, thus providing information on the secure use of this particular cocktail in areas where these parasites are sympatric.

Parasite load and risk factors for poor outcome among children with visceral leishmaniasis. A cohort study in Belo Horizonte, Brazil, 2010-2011

Clinical and laboratory risk factors for death from visceral leishmaniasis (VL) are relatively known, but quantitative real-time polymerase chain reaction (qPCR) might assess the role of parasite load in determining clinical outcome. The aim of this study was to identify risk factors, including parasite load in peripheral blood, for VL poor outcome among children. This prospective cohort study evaluated children aged ≤ 12 years old with VL diagnosis at three times: pre-treatment (T0), during treatment (T1) and post-treatment (T2). Forty-eight patients were included and 16 (33.3%) met the criteria for poor outcome. Age ≤ 12 months [relative risk (RR) 3.51; 95% confidence interval (CI) 1.89-6.52], tachydyspnoea (RR 3.46; 95% CI 2.19-5.47), bacterial infection (RR 3.08; 95% CI 1.27-7.48), liver enlargement (RR 3.00; 95% CI 1.44-6.23) and low serum albumin (RR 7.00; 95% CI 1.80-27.24) were identified as risk factors. qPCR was positive in all patients at T0 and the parasite DNA was undetectable in 76.1% of them at T1 and in 90.7% at T2. There was no statistical association between parasite load at T0 and poor outcome.

A comparative cross-sectional study on the prevalence and morbidity of schistosomiasis in a community in northeastern Brazil (1979-2010)

A cross-sectional study on the prevalence and morbidity of schistosomiasis was conducted in the main settlement of the municipality of Alhandra, in the southern coastal region of the state of Paraíba, in 2010. The results of this study were compared with the results of a previous study conducted in the same area in 1979. The systematic sampling per family conglomerate included approximately 10% of the resident population in the urban area of Alhandra. Faecal examinations were performed using the Kato-Katz method. The clinical forms of the disease were classified in accordance with FS Barbosa as Type I - intestinal form, Type II - hepatointestinal form and Type III - hepatosplenic form. The prevalence of the infection in 2010 was 10.05%, whereas in 1979 it was 46.6% among untreated patients. The percentages of the three clinical forms in 2010 were as follows: 95.3% Type I, 4.6% Type II and 0% Type III; in 1979, the percentages were 94.4%, 3% and 2.6% for Types I, II and III, respectively. In 1979, 6.07% of the Biomphalaria glabrata specimens (the intermediate host in this area) excreted cercariae, where in 2010 only 1.27% of the specimens caught excreted the parasite.

The Plasmodium bottleneck: malaria parasite losses in the mosquito vector

Nearly one million people are killed every year by the malaria parasite Plasmodium. Although the disease-causing forms of the parasite exist only in the human blood, mosquitoes of the genus Anopheles are the obligate vector for transmission. Here, we review the parasite life cycle in the vector and highlight the human and mosquito contributions that limit malaria parasite development in the mosquito host. We address parasite killing in its mosquito host and bottlenecks in parasite numbers that might guide intervention strategies to prevent transmission.

The remarkable journey of adaptation of the Plasmodium falciparum malaria parasite to New World anopheline mosquitoes

Plasmodium falciparum originated in Africa, dispersed around the world as a result of human migration and had to adapt to several different indigenous anopheline mosquitoes. Anophelines from the New World are evolutionary distant form African ones and this probably resulted in a more stringent selection of Plasmodium as it adapted to these vectors. It is thought that Plasmodium has been genetically selected by some anopheline species through unknown mechanisms. The mosquito immune system can greatly limit infection and P. falciparum evolved a strategy to evade these responses, at least in part mediated by Pfs47, a highly polymorphic gene. We propose that adaptation of P. falciparum to new vectors may require evasion of their immune system. Parasites with a Pfs47 haplotype compatible with the indigenous mosquito vector would be able to survive and be transmitted. The mosquito antiplasmodial response could be an important determinant of P. falciparum population structure and could affect malaria transmission in the Americas.

Role of cyclooxygenase-2 in Trypanosoma cruzisurvival in the early stages of parasite host-cell interaction

Chagas disease, caused by the intracellular protozoan Trypanosoma cruzi, is a serious health problem in Latin America. During this parasitic infection, the heart is one of the major organs affected. The pathogenesis of tissue remodelling, particularly regarding cardiomyocyte behaviour after parasite infection and the molecular mechanisms that occur immediately following parasite entry into host cells are not yet completely understood. When cells are infected with T. cruzi, they develop an inflammatory response, in which cyclooxygenase-2 (COX-2) catalyses rate-limiting steps in the arachidonic acid pathway. However, how the parasite interaction modulates COX-2 activity is poorly understood. In this study, the H9c2 cell line was used as our model and we investigated cellular and biochemical aspects during the initial 48 h of parasitic infection. Oscillatory activity of COX-2 was observed, which correlated with the control of the pro-inflammatory environment in infected cells. Interestingly, subcellular trafficking was also verified, correlated with the control of Cox-2 mRNA or the activated COX-2 protein in cells, which is directly connected with the assemble of stress granules structures. Our collective findings suggest that in the very early stage of the T. cruzi-host cell interaction, the parasite is able to modulate the cellular metabolism in order to survives.

Angiostrongylus cantonensis infection in molluscs in the municipality of São Gonçalo, a metropolitan area of Rio de Janeiro, Brazil: role of the invasive species Achatina fulica in parasite transmission dynamics

The aim of this study was to analyse the infection dynamics ofAngiostrongylus cantonensisin its possible intermediate hosts over two years in an urban area in the state of Rio de Janeiro where the presence ofA. cantonensis had been previously recorded in molluscs. Four of the seven mollusc species found in the study were exotic.Bradybaena similariswas the most abundant, followed byAchatina fulica, Streptaxissp., Subulina octona, Bulimulus tenuissimus, Sarasinula linguaeformisand Leptinaria unilamellata. Only A. fulicaand B. similariswere parasitised by A. cantonensis and both presented co-infection with other helminths. The prevalence of A. cantonensisin A. fulicawas more than 50% throughout the study. There was an inverse correlation between the population size ofA. fulicaand the prevalence of A. cantonensisand abundance of the latter was negatively related to rainfall. The overall prevalence of A. cantonensisin B. similariswas 24.6%. A. fulicawas the most important intermediary host of A. cantonensisin the studied area andB. similariswas secondary in importance for A. cantonensistransmission dynamics.

Paralisia cerebral tetraplégica e constipação intestinal: avaliação da reeducação intestinal com uso de massagens e dieta laxante

A constipação intestinal afeta 74% dos indivíduos com paralisia cerebral. O objetivo deste estudo foi avaliar resultados das intervenções de enfermagem no tratamento da constipação intestinal associada à paralisia cerebral. Trata-se de um estudo quantitativo, prospectivo e comparativo (antes-depois). A amostra foi composta por 50 pacientes com paralisia cerebral tetraplégica e constipação intestinal. As principais orientações conservadoras foram: consumo diário de alimentos laxantes e óleos vegetais, aumento da ingestão hídrica e execução de manobras intestinais diárias. Houve alívio total ou parcial da constipação em 90% dos participantes, com melhora de aspectos da qualidade de vida, como sono, apetite e irritabilidade, além de diminuição significativa de sangramento retal, fissura anal, retenção voluntária de fezes, choro e dor ao evacuar. Apenas 10% necessitaram de medicações laxantes. Recomenda-se que medidas conservadoras sejam preferencialmente utilizadas para o tratamento da constipação intestinal associada à paralisia cerebral e que medicamentos sejam apenas adjuvantes, quando necessário.

Utilização de bactérias ácido-lácticas isoladas do trato intestinal de tilápia-do-nilo como probiótico

O objetivo deste trabalho foi isolar bactérias ácido-lácticas do intestino de tilápias-do-nilo, e avaliar seu potencial probiótico. Foram isoladas cepas de bactérias ácido-lácticas, e foi avaliada a inibição aos patógenos in vitro. As cepas com os melhores resultados foram identificadas e utilizadas no experimento de colonização do trato intestinal de tilápias-do-nilo, via suplementação na dieta, em delineamento inteiramente ao acaso, com três tratamentos e quatro repetições. Foram avaliados: o total de bactérias, as bactérias ácido-lácticas, Vibrio ssp. e Pseudomonas ssp. A cepa com melhor resultado foi utilizada na infecção experimental, em delineamento inteiramente ao acaso, em esquema fatorial 2x3: dieta suplementada com a cepa e dieta-controle; e os peixes não submetidos à injeção, peixes submetidos à injeção de solução salina e à injeção de Enterococcus durans, com três repetições. Foram avaliados os parâmetros hematológicos. As duas cepas identificadas foram: Lactobacillus plantarum e Lactobacillus brevis, que colonizaram o trato intestinal de tilápias, contudo L. plantarum teve menor número total de bactérias e de Pseudomonas ssp. Foi observado maior número total de eritrócitos, trombócitos, leucócitos, linfócitos, neutrófilos e monócitos, em peixes alimentados com L. plantarum e submetidos à injeção de E. durans. O L. plantarum tem efeito probiótico e melhora o sistema imune das tilápias.

Obstrução intestinal pós-gastroplastia redutora pela técnica de higa para tratamento da obesidade mórbida: aspectos por imagem

OBJETIVO: O objetivo deste estudo foi descrever os principais aspectos radiológicos encontrados nas obstruções intestinais pós-operatórias em pacientes submetidos a derivação gastrintestinal em Y de Roux pela técnica de Higa. MATERIAIS E MÉTODOS: Foram estudados 10 pacientes com obstrução intestinal no pós-operatório de gastroplastia redutora, examinados entre novembro de 2001 e abril de 2006. Os casos foram obtidos em sete instituições hospitalares distintas. RESULTADOS: Nos 10 pacientes, a obstrução ocorreu em alça de delgado, sendo cinco por hérnia interna, três por brida, um por hérnia umbilical e um por intussuscepção gástrica. Em quatro pacientes a obstrução ocorreu precocemente (até o sétimo dia de pós-operatório) e em seis aconteceu tardiamente (entre o terceiro mês e cinco anos de pós-operatório). CONCLUSÃO: Todos os casos de obstrução intestinal ocorreram ao nível do intestino delgado. A hérnia interna foi a causa mais freqüente, seguida de brida. Outras causas foram hérnia umbilical e intussuscepção gástrica.

Avaliação da concordância entre a ultrassonografia transvaginal e a ressonância magnética da pelve na endometriose profunda, com ênfase para o comprometimento intestinal

OBJETIVO: Comparar achados ultrassonográficos e de ressonância magnética na endometriose profunda, com ênfase para o comprometimento intestinal. MATERIAIS E MÉTODOS: Dezoito pacientes entre 23 e 49 anos de idade, com suspeita clínica e exame ginecológico sugestivo de endometriose profunda, foram submetidas a ultrassonografia e ressonância magnética para correlação dos achados. RESULTADOS: A ultrassonografia detectou 40 lesões e a ressonância magnética detectou 53 lesões na pelve. O estudo comparativo entre ultrassonografia e ressonância magnética na detecção das lesões não mostrou diferença estatística significativa (p > 0,19 e p > 0,14, respectivamente). Considerando-se a junção retossigmoide, a ressonância magnética detectou uma lesão (5,6%) e a ultrassonografia apontou quatro lesões (22,2%). Nas lesões retais, a ultrassonografia apontou oito lesões (44,4%) e a ressonância magnética, sete lesões (38,9%). CONCLUSÃO: A concordância entre a ressonância magnética e a ultrassonografia não foi boa na junção retossigmoide e no reto, sendo que a ultrassonografia detectou um número maior de lesões nessas localizações, mas identificou número menor de lesões na pelve. Na análise comparativa global entre os dois métodos na detecção das lesões não houve diferença estatística significativa. O baixo custo, a boa tolerabilidade e o fácil acesso tornam a ultrassonografia instrumento diagnóstico valioso na endometriose profunda.