Platelet-derived exosomes induce endothelial cell apoptosis through peroxynitrite generation: experimental evidence for a novel mechanism of septic vascular dysfunction

Abstract Introduction Several studies link hematological dysfunction to severity of sepsis. Previously we showed that platelet-derived microparticles from septic patients induce vascular cell apoptosis through the NADPH oxidase-dependent release of superoxide. We sought to further characterize the microparticle-dependent vascular injury pathway. Methods During septic shock there is increased generation of thrombin, TNF-α and nitric oxide (NO). Human platelets were exposed for 1 hour to the NO donor diethylamine-NONOate (0.5 μM), lipopolysaccharide (LPS; 100 ng/ml), TNF-α (40 ng/ml), or thrombin (5 IU/ml). Microparticles were recovered through filtration and ultracentrifugation and analyzed by electron microscopy, flow cytometry or Western blotting for protein identification. Redox activity was characterized by lucigenin (5 μM) or coelenterazine (5 μM) luminescence and by 4,5-diaminofluorescein (10 mM) and 2',7'-dichlorofluorescein (10 mM) fluorescence. Endothelial cell apoptosis was detected by phosphatidylserine exposure and by measurement of caspase-3 activity with an enzyme-linked immunoassay. Results Size, morphology, high exposure of the tetraspanins CD9, CD63, and CD81, together with low phosphatidylserine, showed that platelets exposed to NONOate and LPS, but not to TNF-α or thrombin, generate microparticles similar to those recovered from septic patients, and characterize them as exosomes. Luminescence and fluorescence studies, and the use of specific inhibitors, revealed concomitant superoxide and NO generation. Western blots showed the presence of NO synthase II (but not isoforms I or III) and of the NADPH oxidase subunits p22phox, protein disulfide isomerase and Nox. Endothelial cells exposed to the exosomes underwent apoptosis and caspase-3 activation, which were inhibited by NO synthase inhibitors or by a superoxide dismutase mimetic and totally blocked by urate (1 mM), suggesting a role for the peroxynitrite radical. None of these redox properties and proapoptotic effects was evident in microparticles recovered from platelets exposed to thrombin or TNF-α. Conclusion We showed that, in sepsis, NO and bacterial elements are responsible for type-specific platelet-derived exosome generation. Those exosomes have an active role in vascular signaling as redox-active particles that can induce endothelial cell caspase-3 activation and apoptosis by generating superoxide, NO and peroxynitrite. Thus, exosomes must be considered for further developments in understanding and treating vascular dysfunction in sepsis.

Accuracy of endoscopic ultrasound-guided fine-needle aspiration in the suspicion of pancreatic metastases

Abstract Background Metastases to the pancreas are rare, and usually mistaken for primary pancreatic cancers. This study aimed to describe the histology results of solid pancreatic tumours obtained by endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) for diagnosis of metastases to the pancreas. Methods In a retrospective review, patients with pancreatic solid tumours and history of previous extrapancreatic cancer underwent EUS-FNA from January/1997 to December/2010. Most patients were followed-up until death and some of them were still alive at the end of the study. The performance of EUS-FNA for diagnosis of pancreatic metastases was analyzed. Symptoms, time frame between primary tumour diagnosis and the finding of metastases, and survival after diagnosis were also analyzed. Results 37 patients underwent EUS-FNA for probable pancreas metastases. Most cases (65%) presented with symptoms, especially upper abdominal pain (46%). Median time between detection of the first tumour and the finding of pancreatic metastases was 36 months. Metastases were confirmed in 32 (1.6%) cases, 30 of them by EUS-FNA, and 2 by surgery. Other 5 cases were non-metastatic. Most metastases were from lymphoma, colon, lung, and kidney. Twelve (32%) patients were submitted to surgery. Median survival after diagnosis of pancreatic metastases was 9 months, with no difference of survival between surgical and non-surgical cases. Sensitivity, specificity, positive and negative predictive values, and accuracy of EUS-FNA with histology analysis of the specimens for diagnosis of pancreatic metastases were, respectively, 93.8%, 60%, 93.8%, 60% and 89%. Conclusion EUS-FNA with histology of the specimens is a sensitive and accurate method for definitive diagnosis of metastatic disease in patients with a previous history of extrapancreatic malignancies.

Prevalence and risk factors of Hepatitis C virus infection in Brazil, 2005 through 2009: a cross-sectional study

Abstract Background Hepatitis C chronic liver disease is a major cause of liver transplant in developed countries. This article reports the first nationwide population-based survey conducted to estimate the seroprevalence of HCV antibodies and associated risk factors in the urban population of Brazil. Methods The cross sectional study was conducted in all Brazilian macro-regions from 2005 to 2009, as a stratified multistage cluster sample of 19,503 inhabitants aged between 10 and 69 years, representing individuals living in all 26 State capitals and the Federal District. Hepatitis C antibodies were detected by a third-generation enzyme immunoassay. Seropositive individuals were retested by Polymerase Chain Reaction and genotyped. Adjusted prevalence was estimated by macro-regions. Potential risk factors associated with HCV infection were assessed by calculating the crude and adjusted odds ratios, 95% confidence intervals (95% CI) and p values. Population attributable risk was estimated for multiple factors using a case–control approach. Results The overall weighted prevalence of hepatitis C antibodies was 1.38% (95% CI: 1.12%–1.64%). Prevalence of infection increased in older groups but was similar for both sexes. The multivariate model showed the following to be predictors of HCV infection: age, injected drug use (OR = 6.65), sniffed drug use (OR = 2.59), hospitalization (OR = 1.90), groups socially deprived by the lack of sewage disposal (OR = 2.53), and injection with glass syringe (OR = 1.52, with a borderline p value). The genotypes 1 (subtypes 1a, 1b), 2b and 3a were identified. The estimated population attributable risk for the ensemble of risk factors was 40%. Approximately 1.3 million individuals would be expected to be anti-HCV-positive in the country. Conclusions The large estimated absolute numbers of infected individuals reveals the burden of the disease in the near future, giving rise to costs for the health care system and society at large. The known risk factors explain less than 50% of the infected cases, limiting the prevention strategies. Our findings regarding risk behaviors associated with HCV infection showed that there is still room for improving strategies for reducing transmission among drug users and nosocomial infection, as well as a need for specific prevention and control strategies targeting individuals living in poverty.

Management of diabetes mellitus and associated cardiovascular risk factors in Brazil – the Brazilian study on the practice of diabetes care

Abstract Background The Brazilian Study on the Practice of Diabetes Care main objective was to provide an epidemiological profile of individuals with type 1 and 2 diabetes mellitus (DM) in Brazil, concerning therapy and adherence to international guidelines in the medical practice. Methods This observational, cross-sectional, multicenter study collected and analyzed data from individuals with type 1 and 2 DM attending public or private clinics in Brazil. Each investigator included the first 10 patients with type 2 DM who visited his/her office, and the first 5 patients with type 1 DM. Results A total of 1,358 patients were analyzed; 375 (27.6%) had type 1 and 983 (72.4%) had type 2 DM. Most individuals were women, Caucasian, and private health care users. High prevalence rates of hypertension, dyslipidemia and central obesity were observed, particularly in type 2 DM. Only 7.3% and 5.1% of the individuals with types 1 and 2 DM, respectively, had optimal control of blood pressure, plasma glucose and lipids. The absence of hypertension and female sex were associated with better control of type 1 DM and other cardiovascular risk factors. In type 2 DM, older age was also associated with better control. Conclusions Female sex, older age, and absence of hypertension were associated with better metabolic control. An optimal control of plasma glucose and other cardiovascular risk factors are obtained only in a minority of individuals with diabetes. Local numbers, compared to those from other countries are worse.

Automated system of seedling image analysis (SVIS) and electrical conductivity to assess sun hemp seed vigor

The development of new procedures for quickly obtaining accurate information on the physiological potential of seed lots is essential for developing quality control programs for the seed industry. In this study, the effectiveness of an automated system of seedling image analysis (Seed Vigor Imaging System - SVIS) in determining the physiological potential of sun hemp seeds and its relationship with electrical conductivity tests, were evaluated. SVIS evaluations were performed three and four days after sowing and data on the vigor index and the length and uniformity of seedling growth were collected. The electrical conductivity test was made on 50 seed replicates placed in containers with 75 mL of deionised water at 25 ºC and readings were taken after 1, 2, 4, 8 and 16 hours of imbibition. Electrical conductivity measurements at 4 or 8 hours and the use of the SVIS on 3-day old seedlings can effectively detect differences in vigor between different sun hemp seed lots.

Percepção dos pais sobre a habilidade de atenção auditiva de seu filho com fissura labiopalatina: estudo retrospectivo

INTRODUÇÃO: Para processar e decodificar o estímulo acústico são necessários mecanismos cognitivos e neurofisiológicos. O estímulo auditivo sofre influências de fatores cognitivos de nível mais alto, tais como a memória, atenção e aprendizagem. A privação sensorial ocasionada por perda auditiva do tipo condutiva, frequente na população com fissura labiopalatina, pode afetar várias funções cognitivas - dentre elas a atenção, além de prejudicar os desempenhos escolares, linguísticos e interpessoais. OBJETIVO: Verificar a percepção dos pais de crianças com fissura labiopalatina sobre a atenção auditiva de seus filhos. MÉTODO: Estudo retrospectivo de crianças com qualquer tipo de fissura labiopalatina, sem qualquer síndrome genética associada cujos pais responderam a um questionário pertinente sobre a habilidade de atenção auditiva. RESULTADOS: 44 são do gênero masculino e 26 do gênero feminino, 35,71% das respostas foram afirmativas para a presença de perda auditiva e 71,43% para infecções otológicas. CONCLUSÃO: A maioria dos pais entrevistados apontou pelo menos um dos comportamentos relacionados à atenção contidos no questionário, indicando que a presença de fissura labiopalatina pode estar relacionada com dificuldades quanto à atenção auditiva.

Desfechos intra-hospitalares e preditores de mortalidade no infarto agudo do miocárdio com choque cardiogênico tratados por angioplastia primária: dados do registro InCor

INTRODUÇÃO: O choque cardiogênico é a maior causa de morte em pacientes com infarto agudo do miocárdio com supradesnivelamento do segmento de ST (IAMCSST). O presente estudo avaliou pacientes com IAMCSST e choque cardiogênico submetidos a intervenção coronária percutânea primária com o objetivo de estabelecer seu perfil e os preditores de mortalidade hospitalar. MÉTODOS: Registro unicêntrico, incluindo 100 pacientes avaliados no período de 2001 a 2009 quanto a características clínicas, angiográficas e do procedimento, e a desfechos intra-hospitalares. Por análise multivariada foram determinados preditores independentes da mortalidade hospitalar. RESULTADOS: Com relação às características clínicas, foi observada alta prevalência de fatores de risco, sendo a taxa de sucesso angiográfico de 92%, apesar da complexidade das lesões (83,1% do tipo B2/C). A artéria mais acometida foi a descendente anterior (45%), tendo o padrão multiarterial ocorrido em 73% dos casos. A taxa de mortalidade foi de 45%, sendo seus preditores independentes o padrão multiarterial [odds ratio (OR) 2,62; intervalo de confiança de 95% (IC 95%) 1,16-5,90] e o fluxo coronário TIMI < 3 ao final do procedimento (OR 2,11, IC 95% 1,48-3,02). CONCLUSÕES: Os pacientes com IAMCSST complicado por choque cardiogênico apresentaram características clínicas e angiográficas de alto risco e, apesar do alto sucesso angiográfico do procedimento, altas taxas de mortalidade. Foram preditores independentes de mortalidade o padrão multiarterial e fluxo TIMI < 3 ao final do procedimento.

Gestão do empréstimo unificado nas bibliotecas da Universidade de São Paulo

Apresenta a implantação do empréstimo unificado nas Bibliotecas do Sistema Integrado de Bibliotecas da Universidade de São Paulo (SIBiUSP), coordenado pelo Grupo de Gestão do Empréstimo Unificado. Introduz o cenário inicial das bibliotecas da Universidade, destacando a autonomia para definir políticas locais. Descreve as iniciativas isoladas que culminaram com unificação da política do empréstimo utilizada por todas as bibliotecas. Estabelece comparação entre os procedimentos adotados antes e depois da implantação e conclui relatando os benefícios obtidos pelos 138 mil usuários das bibliotecas USP e na padronização de procedimentos e rotinas adotadas pelas bibliotecas em seus 1,7 milhões de empréstimos domiciliares.

Analysis of Electrostatic Turbulence Drive in Texas Helimak.

Plasma turbulence and particle transport in Texas Helimak change with the radial electric field profile modified by an external voltage bias. When the bias is positive, the turbulence shows enhanced level and broadband spectra with extreme events, similar to the turbulence in tokamak scrape-‐off layer. However, negative bias reduces the turbulence level and decreases the spectrum widths. Moreover, for negative biased shots, the particle transport is strongly affected by a wave particle resonant interaction. On the other hand, for positive bias values, the plasma presents a transport barrier in the reversed shear flow region.

Desenvolvimento de modelo matemático de otimização logística para o transporte multimodal de safras agrícolas pelo corredor centro-oeste

Este artigo apresenta um modelo matemático de otimização logística para o transporte multimodal de safras agrícolas pelo corredor Centro-Oeste. Tal ferramenta foi desenvolvida no contexto de três amplos projetos de pesquisa financiados pela FINEP e executados por um grupo de universidades. O modelo, conhecido genericamente como Modelo de fluxo de Custo Mínimo Multiproduto, considera a otimização de fluxos em rede, para os produtos açúcar, álcool, milho, soja, óleo de soja, farelo de soja e trigo. O modelo proposto para estimativa dos fluxos inter-regionais mostrou-se uma ferramenta factível para fins de avaliação do potencial de utilização da multimodalidade. A análise destes resultados gera importantes subsídios para a seleção dos locais com potencial para instalação de mecanismos e equipamentos de transferência de cargas, além de auxiliar no dimensionamento dessas infraestruturas. Também é um resultado importante do ferramental desenvolvido a identificação das zonas de cargas que apresentam potencial captável pelas ferrovias, hidrovias e dutovias, ou seja, possibilita a identificação das regiões que revelam potencial para uso da multimodalidade.

Membrane interactions of S100A12 (calgranulin C)

S100A12 (Calgranulin C) is a small acidic calcium-binding peripheral membrane protein with two EF-hand structural motifs. It is expressed in macrophages and lymphocytes and highly up-regulated in several human inflammatory diseases. In pigs, S100A12 is abundant in the cytosol of granulocytes, where it is believed to be involved in signal modulation of inflammatory process. In this study, we investigated the interaction of the porcine S100A12 with phospholipid bilayers and the effect that ions (Ca2+, Zn2+ or both together) have in modifying protein-lipid interactions. More specifically, we intended to address issues such as: (1) is the protein-membrane interaction modulated by the presence of ions? (2) is the protein overall structure affected by the presence of the ions and membrane models simultaneously? (3) what are the specific conformational changes taking place when ions and membranes are both present? (4) does the protein have any kind of molecular preferences for a specific lipid component? To provide insight into membrane interactions and answer those questions, synchrotron radiation circular dichroism spectroscopy, fluorescence spectroscopy, and surface plasmon resonance were used. The use of these combined techniques demonstrated that this protein was capable of interacting both with lipids and with ions in solution, and enabled examination of changes that occur at different levels of structure organization. The presence of both Ca2+ and Zn2+ ions modify the binding, conformation and thermal stability of the protein in the presence of lipids. Hence, these studies examining molecular interactions of porcine S100A12 in solution complement the previously determined crystal structure information on this family of proteins, enhancing our understanding of its dynamics of interaction with membranes.