Cardiac-Oxidized Antigens Are Targets of Immune Recognition by Antibodies and Potential Molecular Determinants in Chagas Disease Pathogenesis

Trypanosoma cruzi elicits reactive oxygen species (ROS) of inflammatory and mitochondrial origin in infected hosts. In this study, we examined ROS-induced oxidative modifications in the heart and determined whether the resultant oxidized cardiac proteins are targets of immune response and of pathological significance in Chagas disease. Heart biopsies from chagasic mice, rats and human patients exhibited, when compared to those from normal controls, a substantial increase in protein 4-hydroxynonenal (4-HNE), malondialdehyde (MDA), carbonyl, and 3-nitrotyrosine (3-NT) adducts. To evaluate whether oxidized proteins gain antigenic properties, heart homogenates or isolated cardiomyocytes were oxidized in vitro and one- or two-dimensional gel electrophoresis (2D-GE)/Western blotting (WB) was performed to investigate the proteomic oxidative changes and recognition of oxidized proteins by sera antibodies in chagasic rodents (mice, rats) and human patients. Human cardiomyocytes exhibited LD50 sensitivity to 30 µM 4-HNE and 100 µM H2O2 at 6 h and 12 h, respectively. In vitro oxidation with 4-HNE or H2O2 resulted in a substantial increase in 4-HNE- and carbonyl-modified proteins that correlated with increased recognition of cardiac (cardiomyocytes) proteins by sera antibodies of chagasic rodents and human patients. 2D-GE/Western blotting followed by MALDI-TOF-MS/MS analysis to identify cardiac proteins that were oxidized and recognized by human chagasic sera yielded 82 unique proteins. We validated the 2D-GE results by enzyme-linked immunosorbent assay (ELISA) and WB and demonstrated that oxidation of recombinant titin enhanced its immunogenicity and recognition by sera antibodies from chagasic hosts (rats and humans). Treatment of infected rats with phenyl-α-tert-butyl nitrone (PBN, antioxidant) resulted in normalized immune detection of cardiac proteins associated with control of cardiac pathology and preservation of heart contractile function in chagasic rats. We conclude that ROS-induced, cardiac-oxidized antigens are targets of immune recognition by antibodies and molecular determinants for pathogenesis during Chagas disease

Assessment of predictive models for chlorophyll-a concentration of a tropical lake

<p>Abstract</p> <p>Background</p> <p>This study assesses four predictive ecological models; Fuzzy Logic (FL), Recurrent Artificial Neural Network (RANN), Hybrid Evolutionary Algorithm (HEA) and multiple linear regressions (MLR) to forecast chlorophyll- a concentration using limnological data from 2001 through 2004 of unstratified shallow, oligotrophic to mesotrophic tropical Putrajaya Lake (Malaysia). Performances of the models are assessed using Root Mean Square Error (RMSE), correlation coefficient (r), and Area under the Receiving Operating Characteristic (ROC) curve (AUC). Chlorophyll-a have been used to estimate algal biomass in aquatic ecosystem as it is common in most algae. Algal biomass indicates of the trophic status of a water body. Chlorophyll- a therefore, is an effective indicator for monitoring eutrophication which is a common problem of lakes and reservoirs all over the world. Assessments of these predictive models are necessary towards developing a reliable algorithm to estimate chlorophyll- a concentration for eutrophication management of tropical lakes.</p> <p>Results</p> <p>Same data set was used for models development and the data was divided into two sets; training and testing to avoid biasness in results. FL and RANN models were developed using parameters selected through sensitivity analysis. The selected variables were water temperature, pH, dissolved oxygen, ammonia nitrogen, nitrate nitrogen and Secchi depth. Dissolved oxygen, selected through stepwise procedure, was used to develop the MLR model. HEA model used parameters selected using genetic algorithm (GA). The selected parameters were pH, Secchi depth, dissolved oxygen and nitrate nitrogen. RMSE, r, and AUC values for MLR model were (4.60, 0.5, and 0.76), FL model were (4.49, 0.6, and 0.84), RANN model were (4.28, 0.7, and 0.79) and HEA model were (4.27, 0.7, and 0.82) respectively. Performance inconsistencies between four models in terms of performance criteria in this study resulted from the methodology used in measuring the performance. RMSE is based on the level of error of prediction whereas AUC is based on binary classification task.</p> <p>Conclusions</p> <p>Overall, HEA produced the best performance in terms of RMSE, r, and AUC values. This was followed by FL, RANN, and MLR.</p

Propriedades Mecânicas de Epóxis Utilizadas no Recobrimento Interno de Oleodutos e Gasodutos Mechanical Properties of Epoxy for Internal Coatings of Pipelines

Propriedades mecânicas de recobrimentos a base de epóxi para aplicação in situ em dutos de transporte na indústria petrolífera foram avaliadas. Os recobrimentos foram aplicados sobre substratos de aço carbono submetidos a dois diferentes padrões de tratamento superficial (ST3 e SA2&frac12;) e avaliou-se a influência do mesmo em propriedades tais como adesão, impacto, dureza e desgaste do material. Observou-se que a falha de adesão ocorre por diferentes mecanismos dependendo do material do recobrimento e da preparação da superfície. Desgaste ocorre basicamente por mecanismo abrasivo embora se tenha observado desgaste adesivo significativo em alguns casos, dependendo do material e do padrão de tratamento da superfície. Resultados de microdureza indicaram, como esperado, ser esta uma propriedade inerente ao material, não sendo afetada pelo padrão de limpeza da superfície<br>Mechanical properties of epoxy based coating materials suitable for in-situ internal application in gas/oil pipelines were studied. Coatings were applied on carbon steel substrates submitted to two different surface preparation procedures (ST3 and SA2&frac12;) and the effect from surface treatment on coating adhesion, impact, hardness and wear was evaluated. It was found that adhesion failure occurs through distinct mechanisms depending on coating material and surface preparation. Wear occurs mostly by abrasive mechanism, although adhesive wear was found to be significant in some cases also depending on material and surface. Microhardness, as expected, was found to be a coating property not unaffected by surface treatment

Cardiac gene expression and systemic cytokine profile are complementary in a murine model of post-ischemic heart failure

After myocardial infarction (MI), activation of the immune system and inflammatory mechanisms, among others, can lead to ventricular remodeling and heart failure (HF). The interaction between these systemic alterations and corresponding changes in the heart has not been extensively examined in the setting of chronic ischemia. The main purpose of this study was to investigate alterations in cardiac gene and systemic cytokine profile in mice with post-ischemic HF. Plasma was tested for IgM and IgG anti-heart reactive repertoire and inflammatory cytokines. Heart samples were assayed for gene expression by analyzing hybridization to AECOM 32k mouse microarrays. Ischemic HF significantly increased the levels of total serum IgM (by 5.2-fold) and total IgG (by 3.6-fold) associated with a relatively high content of anti-heart specificity. A comparable increase was observed in the levels of circulating pro-inflammatory cytokines such as IL-1&#946; (3.8X) and TNF-&#945; (6.0X). IFN-&#947; was also increased by 3.1-fold in the MI group. However, IL-4 and IL-10 were not significantly different between the MI and sham-operated groups. Chemokines such as MCP-1 and IL-8 were 1.4- and 13-fold increased, respectively, in the plasma of infarcted mice. We identified 2079 well annotated unigenes that were significantly regulated by post-ischemic HF. Complement activation and immune response were among the most up-regulated processes. Interestingly, 21 of the 101 quantified unigenes involved in the inflammatory response were significantly up-regulated and none were down-regulated. These data indicate that post-ischemic heart remodeling is accompanied by immune-mediated mechanisms that act both systemically and locally

Cardiac gene expression and systemic cytokine profile are complementary in a murine model of post-ischemic heart failure

Submitted by Sandra Infurna ([email protected]) on 2018-12-09T17:53:33Z No. of bitstreams: 1 hugocc2_farianeto_etal_IOC_2010.pdf: 1188349 bytes, checksum: 8fd97f76f99ffad2ede3b0be771b7d34 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2018-12-09T18:10:01Z (GMT) No. of bitstreams: 1 hugocc2_farianeto_etal_IOC_2010.pdf: 1188349 bytes, checksum: 8fd97f76f99ffad2ede3b0be771b7d34 (MD5)Made available in DSpace on 2018-12-09T18:10:01Z (GMT). No. of bitstreams: 1 hugocc2_farianeto_etal_IOC_2010.pdf: 1188349 bytes, checksum: 8fd97f76f99ffad2ede3b0be771b7d34 (MD5) Previous issue date: 2010Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Rio de Janeiro, RJ, Brasil.Centro de Ensino e Pesquisa, Hospital Pró-Cardíaco, Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Fisiologia e Farmacodinâmica. Rio de Janeiro, RJ, Brasil.Universidade Federal Fluminense. Instituto de Imunologia. Departamento de Imunobiologia. Niterói, RJ. Brasil.Albert Einstein College of Medicine. Bronx, NY, USA.Albert Einstein College of Medicine. Bronx, NY, USA.Centro de Ensino e Pesquisa, Hospital Pró-Cardíaco, Rio de Janeiro, RJ, Brasil.Albert Einstein College of Medicine. Bronx, NY, USA.Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biofísica Carlos Chagas Filho. Rio de Janeiro, RJ, Brasil / Albert Einstein College of Medicine. Bronx, NY, USA.After myocardial infarction (MI), activation of the immune system and inflammatory mechanisms, among others, can lead to ventricular remodeling and heart failure (HF). The interaction between these systemic alterations and corresponding changes in the heart has not been extensively examined in the setting of chronic ischemia. The main purpose of this study was to investigate alterations in cardiac gene and systemic cytokine profile in mice with post-ischemic HF. Plasma was tested for IgM and IgG anti-heart reactive repertoire and inflammatory cytokines. Heart samples were assayed for gene expression by analyzing hybridization to AECOM 32k mouse microarrays. Ischemic HF significantly increased the levels of total serum IgM (by 5.2-fold) and total IgG (by 3.6-fold) associated with a relatively high content of anti-heart specificity. A comparable increase was observed in the levels of circulating pro-inflammatory cytokines such as IL-1β (3.8X) and TNF-α (6.0X). IFN-γ was also increased by 3.1-fold in the MI group. However, IL-4 and IL-10 were not significantly different between the MI and sham-operated groups. Chemokines such as MCP-1 and IL-8 were 1.4- and 13-fold increased, respectively, in the plasma of infarcted mice. We identified 2079 well annotated unigenes that were significantly regulated by post-ischemic HF. Complement activation and immune response were among the most up-regulated processes. Interestingly, 21 of the 101 quantified unigenes involved in the inflammatory response were significantly up-regulated and none were down-regulated. These data indicate that post-ischemic heart remodeling is accompanied by immune-mediated mechanisms that act both systemically and locally