Epigenetic Markers Associated with Schistosomiasis.

It is important to consider the use of the epigenome as source of complementary data for genome knowledge, which is suitable for the diagnosis of schistosomiasis. Usually, a laboratory diagnosis of schistosomiasis is performed by means of 1. Egg detection in the stool or urine by microscopy remains with limited sensitivity; 2. Immunological screening, in which positivity persists after treatment, and 3. Molecular appraisals prevail over the disadvantages of the currently used methods. In this sense, molecular methodologies are being developed based on epigenetic biomarkers, aiming to improve the diagnosis of the disease and clinical treatment as early as possible to prevent the occurrence of serious liver damage.

Epididymis tuberculosis: Case report and brief review of the literature.

Tuberculosis (TB) is a secular disease caused by a bacillus, highly prevalent in Brazil. The genito-urinary tract involvement is rare, with the epididymis the most affected location. Treatment usually involves the combination of 3-4 drugs for TB for 6 months and surgery can be useful in complications.

Potential immunological markers for diagnosis of human strongyloidiasis using heterologous antigens.

Strongyloides venezuelensis is a parasitic nematode of rodents that is frequently used to obtain heterologous antigens for immunological diagnosis of human strongyloidiasis. The aim of this study was to identify antigens from filariform larvae of S. venezuelensis for immunodiagnosis of human strongyloidiasis. Soluble and membrane fractions from filariform larvae of S. venezuelensis were obtained in phosphate saline (SS and SM) and in Tris-HCl buffer (TS and TM), and were analysed by Western blotting. Different antigenic components were recognized by IgG antibodies from the sera of strongyloidiasis patients. Highest recognition was observed for a 30-40 kDa mass range present in all antigenic fractions. The band encompassing this mass range was then excised and subjected to mass spectrometry for protein identification. Immunoreactive proteins identified in the soluble fractions corresponded to metabolic enzymes, whereas cytoskeletal proteins and galectins were more abundant in the membrane fractions. These results represent the first approach towards identification of S. venezuelensis antigens for use in immunodiagnostic assays for human strongyloidiasis.

Evaluation of EPS extraction protocols from anaerobic sludge for gel-based proteomic studies.

Despite the importance of anaerobic sludge extracellular polymeric substances (EPSs), their characterization is limited to information regarding their chemical classes and molecular size. This work explores the possibility of using proteomic techniques to study the proteins present in this matrix. Thus, this paper compares eight EPS extraction methods regarding extraction yield, protein/carbohydrate ratio, size distribution profile and suitability to sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses. Despite the differences found in quantification and size exclusion chromatography assays, the band profile found for all methods was very similar. Considering the band pattern, extraction time and background level, heating method followed by ammonium sulfate precipitation proved to be the most appropriate method for gel-based analyses of anaerobic sludge EPS proteins.

Protocolo gerenciado de tratamento do potencial doador falecido reduz incidência de parada cardíaca antes do explante dos órgãos / A managed protocol for treatment of deceased potential donors reduces the incidence of cardiac arrest before organ explant

OBJETIVO: Avaliar o efeito da aplicação de um protocolo gerenciado de manutenção de potenciais doadores falecidos de múltiplos órgãos em duas unidades hospitalares. MÉTODOS: Estudo antes (Fase 1)/depois (Fase 2) realizado em dois hospitais gerais que incluiu, consecutivamente, os potenciais doadores ingressados em duas unidades de terapia intensiva. Na Fase 1 (16 meses), os dados foram coletados retrospectivamente e as medidas de manutenção do potencial doador foram instituídas a critério do intensivista. Na Fase 2 (12 meses), a coleta de dados foi prospectiva e a manutenção foi guiada por um protocolo gerenciado. As duas fases foram comparadas entre si de acordo com variáveis demográficas, variáveis fisiológicas no diagnóstico da morte encefálica e ao final do processo, tempo necessário para realização do exame confirmatório de morte encefálica e final do processo, aderência aos conjuntos de medidas essenciais de manutenção (pacotes), perdas por parada cardíaca, perdas por negativa familiar, perdas por contraindicação e taxa de conversão de potenciais doadores em doadores reais. Foram aplicados os testes de t-Student e do qui-quadrado, e o valor de p<0,05 foi considerado significativo. RESULTADOS: Identificaram-se 42 potenciais doadores (18 na Fase 1 e 24 na Fase 2). Houve diminuição do tempo entre a primeira exploração clínica e o explante (Fase 1: 35,0±15,5 horas versus Fase 2: 24,6±6,2 horas; p=0,023). Houve aumento na aderência em 10 dos 19 itens essenciais de manutenção, e redução nas perdas por parada cardíaca (Fase 1: 27,8 versus 0% na Fase 2; p=0,006) com aumento de doadores reais (Fase 1: 44,4 versus 75% na Fase 2; p=0,044). Não houve mudança nas perdas por negativa familiar ou por contraindicação médica. CONCLUSÃO: A adoção de um protocolo gerenciado promove a aplicação de medidas essenciais no cuidado do potencial doador falecido e pode reduzir as perdas de potenciais doadores por parada cardíaca.

OBJECTIVE: To assess the effect of the application of a managed protocol for the maintenance care of deceased potential multiple organ donors at two hospitals. METHODS: A before (Phase 1)/after (Phase 2) study conducted at two general hospitals, which included consecutively potential donors admitted to two intensive care units. In Phase 1 (16 months), the data were collected retrospectively, and the maintenance care measures of the potential donors were instituted by the intensivists. In Phase 2 (12 months), the data collection was prospective, and a managed protocol was used for maintenance care. The two phases were compared in terms of their demographic variables, physiological variables at diagnosis of brain death and the end of the process, time to performance of brain death confirmatory test and end of the process, adherence to bundles of maintenance care essential measures, losses due to cardiac arrest, family refusal, contraindications, and the conversion rate of potential into actual donors. Student's t- and chi-square tests were used, and p-value < 0.05 was considered to be significant. RESULTS: A total of 42 potential donors were identified (18 in Phase 1 and 24 in Phase 2). The time interval between the first clinical assessment and the recovery decreased in Phase 2 (Phase 1: 35.0±15.5 hours versus Phase 2: 24.6±6.2 hours; p = 0.023). Adherence increased to 10 out of the 19 essential items of maintenance care, and losses due to cardiac arrest also decreased in Phase 2 (Phase 1: 27.8 versus 0% in Phase 2; p = 0.006), while the convertion rate increased (Phase 1: 44.4 versus 75% in Phase 2; p = 0.044). The losses due to family refusal and medical contraindication did not vary. CONCLUSION: The adoption of a managed protocol focused on the application of essential measures for the care of potential deceased donors might reduce the loss of potential donors due to cardiac arrest.

A managed protocol for treatment of deceased potential donors reduces the incidence of cardiac arrest before organ explant.

OBJECTIVE: To assess the effect of the application of a managed protocol for the maintenance care of deceased potential multiple organ donors at two hospitals. METHODS: A before (Phase 1)/after (Phase 2) study conducted at two general hospitals, which included consecutively potential donors admitted to two intensive care units. In Phase 1 (16 months), the data were collected retrospectively, and the maintenance care measures of the potential donors were instituted by the intensivists. In Phase 2 (12 months), the data collection was prospective, and a managed protocol was used for maintenance care. The two phases were compared in terms of their demographic variables, physiological variables at diagnosis of brain death and the end of the process, time to performance of brain death confirmatory test and end of the process, adherence to bundles of maintenance care essential measures, losses due to cardiac arrest, family refusal, contraindications, and the conversion rate of potential into actual donors. Student's t- and chi-square tests were used, and p-value < 0.05 was considered to be significant. RESULTS: A total of 42 potential donors were identified (18 in Phase 1 and 24 in Phase 2). The time interval between the first clinical assessment and the recovery decreased in Phase 2 (Phase 1: 35.0±15.5 hours versus Phase 2: 24.6±6.2 hours; p = 0.023). Adherence increased to 10 out of the 19 essential items of maintenance care, and losses due to cardiac arrest also decreased in Phase 2 (Phase 1: 27.8 versus 0% in Phase 2; p = 0.006), while the convertion rate increased (Phase 1: 44.4 versus 75% in Phase 2; p = 0.044). The losses due to family refusal and medical contraindication did not vary. CONCLUSION: The adoption of a managed protocol focused on the application of essential measures for the care of potential deceased donors might reduce the loss of potential donors due to cardiac arrest.

Schistosoma mansoni: functional proteasomes are required for development in the vertebrate host.

Proteasomes are multi-subunit proteases involved in several mechanisms and thought to contribute to the regulation of cellular homeostasis. Here, we report for the first time biochemical evidence for the existence of a ubiquitin-proteasome proteolytic pathway in this parasite. Proteasomes from both cercariae and adult worms exhibited a high preference for hydrolysis of the substrate Suc-LLVY-AMC, although in the cercariae extract the rate of hydrolysis was 50% lower when compared to adult worms extracts. The same difference in proteasome activities was observed when endogenous proteins were broken down in the presence of ATP and ubiquitin. Additionally, accumulation of high molecular weight conjugates was observed when cercariae were pre-incubated with proteasome inhibitors. Finally, we present evidence that during experimental schistosomiasis, proteasome inhibitors were able to reduce the number of lung stage schistosomula, reduce the worm burden and consequently decrease the egg output in infected mice.