Efeito da berinjela sobre os lípides plasmáticos, a peroxidação lipídica e a reversão da disfunção endotelial na hipercolesterolemia experimental

OBJETIVO: Estudar o efeito do suco da berinjela sobre os lípides plasmáticos, o colesterol tecidual, a peroxidação lipídica das LDL nativas, oxidadas e da parede arterial e o relaxamento dependente do endotélio, em coelhos hipercolesterolêmicos. MÉTODOS: Coelhos foram separados em grupos controle (GC), hipercolesterolêmico (GH) e berinjela (GB), (n=10). Os animais do GC foram alimentados com ração normal, o GH e o GB com ração acrescentada de colesterol (0,5%) e gordura de babaçu (10%) durante 30 dias. Ao GB acrescentou-se suco de berinjela, nos últimos 15 dias do experimento. Os lípides plasmáticos foram medidos através de kits enzimáticos, a peroxidação lipídica pela dosagem do malondialdeído (MDA) e o relaxamento dependente do endotélio, por curvas de concentração efeito pela acetilcolina e nitroprussiato. RESULTADOS: O peso dos animais foi menor no GB em relação ao GC e GH (p<0,05). O colesterol total plasmático, as LDL e os triglicérides que se elevaram no GH, reduziram-se em 19%, 29% e 38%, respectivamente no GB (p<0,05). O colesterol tecidual foi menor no GB (50%) em relação ao GH (p<0,05). Nos animais do GB ocorreu redução significante do teor de MDA nas LDL nativas (56%) e oxidadas (22%), assim como na parede arterial (30%) (p<0,05). O relaxamento máximo dependente do endotélio, , elevou-se significantemente em 28% no GB em relação ao GH (p<0,05). CONCLUSÃO: O suco de berinjela administrado a coelhos hipercolesterolêmicos reduziu significantemente o peso corpóreo, o colesterol total, as LDL plasmáticas, os triglicérides, o colesterol tecidual, a peroxidação lipídica das LDL nativas, oxidadas e da parede arterial, assim como aumentou o relaxamento dependente do endotélio.

Efeitos da atorvastatina, fluvastatina, pravastatina e simvastatina sobre a função endotelial, a peroxidação lipídica e a aterosclerose aórtica em coelhos hipercolesterolêmicos

OBJETIVO: Comparar os efeitos da atorvastatina, fluvastatina, pravastatina e simvastatina sobre a função endotelial, a aterosclerose aórtica e o teor de malonodialdeído (MDA) nas LDL nativas, oxidadas e na parede arterial de coelhos hipercolesterolêmicos, depois que as doses destas estatinas foram ajustadas para reduzir o colesterol total plasmático a valores similares. MÉTODOS: Coelhos machos, foram separados em grupos de 10 animais (n=10), chamados hipercolesterolêmico (controle), atorvastatina, fluvastatina, pravastatina e normal. A exceção do grupo normal, os animais foram alimentados com ração padrão acrescida de colesterol a 0,5% e óleo de coco a 2% durante 45 dias. As drogas foram administradas a partir do 15º dia do início do experimento e no 30º dia, as doses foram ajustadas, através do controle do colesterol plasmático, para obter valores semelhantes em cada grupo. Ao final do experimento foi dosado o colesterol plasmático e as lipoproteinas e retirado um segmento de aorta torácica para estudo da função endotelial, da peroxidação lipídica e exame histológico para medida da aterosclerose aórtica. RESULTADOS: As estatinas reduziram significantemente o colesterol total plasmático, as LDL-colesterol e a aterosclerose aórtica. O teor de MDA também foi significantemente reduzido nas LDL nativas e oxidadas, assim como na parede arterial. O relaxamento-dependente do endotélio foi significantemente maior no grupo tratado em comparação ao hipercolesterolêmico. CONCLUSÃO: As estatinas, em doses ajustadas, tiveram efeito significante e similar em reduzir a peroxidação lipídica nas LDL e na parede arterial, na regressão da aterosclerose aórtica e na reversão da disfunção endotelial.

Producción de conocimientos aplicables a una solución tecnológica, a escala de laboratorio o equivalente

Entre los problemas comerciales más importantes que el maní de origen argentino enfrenta en el mercado internacional se destaca la amplia variación en los volúmenes ofertados anualmente, debido a las oscilaciones en la producción y el rendimiento, lo cual dificulta satisfacer plenamente la demanda internacional creciente de maní confitería. Argentina tiene una capacidad exportadora de alrededor de 400.000 toneladas por año. Sin embargo, ante una oferta decreciente de los principales competidores en el mercado internacional, EE.UU y China, la demanda queda insatisfecha, por lo cual nuestro país podría aumentar sus exportaciones en no menos de un 25%, lo que representaría no menos de 150 millones de dólares, que actualmente se dejan de percibir. Problemas de producción y el avance de nuevas tecnologías de cultivo están cambiando el panorama en la producción cordobesa y nacional del maní. Por una parte, la principal región productora argentina se está ampliando hacia el sur de la Provincia de Córdoba y, también, se está desarrollando el cultivo en el noroeste argentino (NOA), ocupándose ambientes sustancialmente distintos a la de la zona núcleo tradicional. Soave (1997), puntualiza que la obtención de nuevas variedades, que amplíen el estrecho panorama varietal existente e incorporen caracteres que respondan a las condiciones locales debe ser un objetivo prioritario y permanente. La literatura proporciona evidencia clara que las especies silvestres del género Arachis son fuentes potenciales de altos niveles de resistencia/tolerancia a enfermedades y sequía. Por otro lado, Fávero (2004) ha demostrado la posibilidad cierta de transferencia de atributos genéticos presente en estas especies al maní cultivado, mediante la construcción de anfidiploides sintéticos de cruzamiento de una especie silvestre con genoma A y otra con genoma B. La cuantificación del contenido de MDA en hoja es un buen parámetro para estimar la resistencia o susceptibilidad de un genotipo a estrés hídrico. Asimismo, estudios recientes indicaron que el uso del área foliar (SLA), el contenido de clorofila (SCMR) y el índice de cosecha, los cuales son sencillos de medir, se correlacionan significativamente con la eficiencia de transpiración y tienen una considerable variación genética en maní. Estos atributos permiten seleccionar genotipos resistentes/tolerantes a sequía. Innovar en el campo de la biotecnología a partir de construcciones genéticas alternativas es una estrategia que permitiría obtener plantas resistentes/tolerantes con la introducción de genes que estén directamente involucrados con los eventos de interés y que ya han sido estudiados en otros organismos. La obtención de plantas con mayor tolerancia a la sequía y/o a enfermedades fúngicas no sólo aseguraría la estabilidad de los rindes en años de escasez hídrica y condiciones climáticas favorables para las enfermedades fúngicas, sino que permitiría extender la frontera productiva a regiones actualmente marginales. Se plantea como objetivo general caracterizar y evaluar fuentes de resistencia/tolerancia a factores bióticos y abióticos, particularmente a Sclerotinia minor, Sclerotinia sclerotiorum y sequía, en gentipos cultivados y silvestres, para transferir los genes de interés mediante técnicas de mejoramiento convencional; y ajustar las condiciones para la regeneración in vito y la transformación génica de variedades elite de maní cultivadas en Argentina.

Efecto de los ácidos grasos poliinsaturados y sus metabolitos en la modulación de factores de transcripción sobre la carcinogénesis. Effects of Polyunsaturatd fatty acids and their metabolites on transcriptional factors modulation in carcinogenesis.

El cáncer se origina por mutaciones, competición y selección natural en células somáticas de tejidos de diferentes órganos,siendo un proceso complejo y multifactorial que ocurre en una secuencia de etapas: iniciación, promoción y progresión (1). Factores hereditarios,genéticos y epigenéticos como los lípidos dietarios, estrés oxidativo, hormonas, pesticidas y otros, influyen tanto en el desarrollo como en la inhibición de esta enfermedad (2). Datos epidemiológicos y experimentales tanto nuestros como de otros laboratorios han demostrado que el consumo de dietas ricas en ácidos grasos de la familia n-3, n-6 o n-9 cambian la fluidez, la actividad de enzimas, el nivel de proteínas y favorecen la formación de moléculas bioactivas derivadas de los lípidos como los eicosanoides y endocanabinoides que modulan el proceso carcinogénico (3-15). Estos derivados lípídicos activan vias de señalización produciendo cambios especificos en la expresión génica, un proceso fundamental durante la transformación neoplásica (1-2).También ha sido demostrado que estos cambios en la expresión génica inducidos por derivados lipídicos modulan funciones en células cancerosas como proliferación y muerte celular, migración y producción de matriz extracelular (16-17). A pesar de estos conocimientos, la identidad de los derivados lipídicos implicados en la modulación de la expresión génica durante la transformación neoplásica asi como los mecanismos utilizados por estas moléculas permanecen aun poco conocidos. HIPOTESIS: En los modelos a utilizar en el presente proyecto, la variación lipídica de las membranas que se induzca por manipulación dietaria deberán generar también variaciones en los eicosanoides . endocanabinoides y otros peróxidos que afecten factores de transcripción nucleares como el p53 y GLI incidiendo en los mecanismos responsables de la muerte y proliferación de células cancerosas. OBJETIVOS: Nos proponemos establecer el impacto de dietas enriquecidas con ácidos grasos de las familias n-3, n-6 o n-9 sobre modelos experimentales in-vivo e in-vitro. Se estudiarán los ácidos grasos de membrana plasmática, la generación de eicosanoides y endocanabinoides derivados de las vias COX y LOX Además se determinará el efecto de los peróxidos en la expresión y actividad de los factores nucleares de transcripción p53 y GLI como mecanismos responsables de la muerte y proliferación celular. MATERIALES Y MÉTODO: Se utilizará un modelo in-vivo de cáncer de mama empleando ratones C57BL6J inducidos con DMBA que se alimentarán con una dieta base semi-sintética suplemetada con diferentes PUFAs (Chia: n-3, Maíz: n-6 y Oleico: n-9 , empleada en estudios previos (8).Modelos in vitro: se utilizarán lineas celulares cancerígenas humanas de mama MCF-7 y MDA, las cuales se tratarán exógenamente con diferentes PUFAS (GLA:n-6,EPA:n-3, Oleico n-9)(9). Se determinarán ácidos grasos de membranas por Cromatografía de gas (CG)(10-11). El análisis de eicosanoides en células tumorales se realizará por HPLC (9-11). Los endocanabinoides por GC-Espectometría de Masa (18).La formación de peróxidos intracelulares se determinará por análisis de Glutation reducido (GSH)(16).La apoptosis se medirá por actividad caspasas y por Citometria de flujo usando Annexina V FICT (19).La expresión celular de Tp53 y GLI se realizará por Western Blot, PCR e inmunohistoquímica (20-21).RESULTADOS ESPERADOS: Se espera que los lípidos añadidos en las dietas de ratones inyectados con DMBA o al medio de cultivo de células tumorales de mama o páncreas modifiquen los ácidos grasos de membrana y sus derivados lipídicos los eicosanoides y endocanabionoides que suponemos afectarán la activación y expresión de factores de transcripción regulando la carcinogénesis. IMPORTANCIA: Diseñar nuevos modelos experimentales para implementar en terapias génicas y aplicar los resultados sobre factores nutricionales que pudieran actuar como inhibidores o promotores del desarrollo del cáncer en humanos.

Marcadores de desequilíbrio redox em sangue de pacientes hipertensos de uma comunidade no Nordeste do Brasil

FUNDAMENTO: Estudos recentes descrevem a participação de espécies reativas de oxigênio e nitrogênio na hipertensão. OBJETIVO: Identificar o desbalanço redox em sangue de hipertensos. MÉTODOS: Superóxido dismutase (SOD), catalase (CAT), glutationa peroxidase (GPx), glutationa (GSH), vitamina C, transferrina, ceruloplasmina, malondialdeído (MDA) e o grupo carbonila, foram quantificados no sangue de 20 hipertensos e 21 controles. Os indivíduos tinham um Índice de Massa Corporal de > 18,5 e < 30 kg/m², glicemia < 100 mg/dL, colesterol sérico < 200 mg/dL, e eram mulheres não fumantes, não grávidas e não lactantes, não usuárias de alopurinol e probucol, e hipertensos em medicação anti-hipertensiva. Todos os indivíduos foram submetidos a um período preparatório de quatro semanas sem álcool, suplementos vitamínicos, dexametasona e paracetamol. RESULTADOS: Níveis reduzidos de CAT (p = 0,013), GSH ( p = 0,003) e MDA (p = 0,014), e altos níveis de GPx (p = 0,001) e ceruloplasmina (p = 0,015) foram obtidos no grupo de hipertensos, em comparação com os controles. Foi verificada uma correlação positiva entre a pressão sistólica e o MDA nos hipertensos e diastólica e CAT nos controles. CONCLUSÃO: Os dados obtidos são sugestivos de que os hipertensos apresentavam desequilíbrio em reações redox, a despeito do possível efeito atenuante de sua medicação anti-hipertensiva.

Tramadol Alleviates Myocardial Injury Induced by Acute Hindlimb Ischemia Reperfusion in Rats

AbstractBackground:Organ injury occurs not only during periods of ischemia but also during reperfusion. It is known that ischemia reperfusion (IR) causes both remote organ and local injuries.Objective:This study evaluated the effects of tramadol on the heart as a remote organ after acute hindlimb IR.Methods:Thirty healthy mature male Wistar rats were allocated randomly into three groups: Group I (sham), Group II (IR), and Group III (IR + tramadol). Ischemia was induced in anesthetized rats by left femoral artery clamping for 3 h, followed by 3 h of reperfusion. Tramadol (20 mg/kg, intravenous) was administered immediately prior to reperfusion. At the end of the reperfusion, animals were euthanized, and hearts were harvested for histological and biochemical examination.Results:The levels of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were higher in Groups I and III than those in Group II (p < 0.05). In comparison with other groups, tissue malondialdehyde (MDA) levels in Group II were significantly increased (p < 0.05), and this increase was prevented by tramadol. Histopathological changes, including microscopic bleeding, edema, neutrophil infiltration, and necrosis, were scored. The total injuryscore in Group III was significantly decreased (p < 0.05) compared with Group II.Conclusion:From the histological and biochemical perspectives, treatment with tramadol alleviated the myocardial injuries induced by skeletal muscle IR in this experimental model.

Using information from telephone panel surveys to predict reasons for refusal

One of the key problems in conducting surveys is convincing people to participate.¦However, it is often difficult or impossible to determine why people refuse. Panel surveys¦provide information from previous waves that can offer valuable clues as to why people¦refuse to participate. If we are able to anticipate the reasons for refusal, then we¦may be able to take appropriate measures to encourage potential respondents to participate¦in the survey. For example, special training could be provided for interviewers¦on how to convince potential participants to participate.¦This study examines different influences, as determined from the previous wave,¦on refusal reasons that were given by the respondents in the subsequent wave of the¦telephone Swiss Household Panel. These influences include socio-demography, social¦inclusion, answer quality, and interviewer assessment of question understanding and¦of future participation. Generally, coefficients are similar across reasons, and¦between-respondents effects rather than within-respondents effects are significant.¦While 'No interest' reasons are easier to predict, the other reasons are more situational. Survey-specific issues are able to distinguish¦different reasons to some extent.

Particulate formulations for the delivery of poly(I:C) as vaccine adjuvant.

Current research and development of antigens for vaccination often center on purified recombinant proteins, viral subunits, synthetic oligopeptides or oligosaccharides, most of them suffering from being poorly immunogenic and subject to degradation. Hence, they call for efficient delivery systems and potent immunostimulants, jointly denoted as adjuvants. Particulate delivery systems like emulsions, liposomes, nanoparticles and microspheres may provide protection from degradation and facilitate the co-formulation of both the antigen and the immunostimulant. Synthetic double-stranded (ds) RNA, such as polyriboinosinic acid-polyribocytidylic acid, poly(I:C), is a mimic of viral dsRNA and, as such, a promising immunostimulant candidate for vaccines directed against intracellular pathogens. Poly(I:C) signaling is primarily dependent on Toll-like receptor 3 (TLR3), and on melanoma differentiation-associated gene-5 (MDA-5), and strongly drives cell-mediated immunity and a potent type I interferon response. However, stability and toxicity issues so far prevented the clinical application of dsRNAs as they undergo rapid enzymatic degradation and bear the potential to trigger undue immune stimulation as well as autoimmune disorders. This review addresses these concerns and suggests strategies to improve the safety and efficacy of immunostimulatory dsRNA formulations. The focus is on technological means required to lower the necessary dosage of poly(I:C), to target surface-modified microspheres passively or actively to antigen-presenting cells (APCs), to control their interaction with non-professional phagocytes and to modulate the resulting cytokine secretion profile.

Lufaxin, a novel factor Xa inhibitor from the salivary gland of the sand fly Lutzomyia longipalpis blocks protease-activated receptor 2 activation and inhibits inflammation and thrombosis in vivo.

OBJECTIVE: Blood-sucking arthropods' salivary glands contain a remarkable diversity of antihemostatics. The aim of the present study was to identify the unique salivary anticoagulant of the sand fly Lutzomyia longipalpis, which remained elusive for decades. METHODS AND RESULTS: Several L. longipalpis salivary proteins were expressed in human embryonic kidney 293 cells and screened for inhibition of blood coagulation. A novel 32.4-kDa molecule, named Lufaxin, was identified as a slow, tight, noncompetitive, and reversible inhibitor of factor Xa (FXa). Notably, Lufaxin's primary sequence does not share similarity to any physiological or salivary inhibitors of coagulation reported to date. Lufaxin is specific for FXa and does not interact with FX, Dansyl-Glu-Gly-Arg-FXa, or 15 other enzymes. In addition, Lufaxin blocks prothrombinase and increases both prothrombin time and activated partial thromboplastin time. Surface plasmon resonance experiments revealed that FXa binds Lufaxin with an equilibrium constant ≈3 nM, and isothermal titration calorimetry determined a stoichiometry of 1:1. Lufaxin also prevents protease-activated receptor 2 activation by FXa in the MDA-MB-231 cell line and abrogates edema formation triggered by injection of FXa in the paw of mice. Moreover, Lufaxin prevents FeCl(3)-induced carotid artery thrombus formation and prolongs activated partial thromboplastin time ex vivo, implying that it works as an anticoagulant in vivo. Finally, salivary gland of sand flies was found to inhibit FXa and to interact with the enzyme. CONCLUSIONS: Lufaxin belongs to a novel family of slow-tight FXa inhibitors, which display antithrombotic and anti-inflammatory activities. It is a useful tool to understand FXa structural features and its role in prohemostatic and proinflammatory events.

Selective and powerful stress gene expression in Arabidopsis in response to malondialdehyde.

The provenance, half-life and biological activity of malondialdehyde (MDA) were investigated in Arabidopsis thaliana. We provide genetic confirmation of the hypothesis that MDA originates from fatty acids containing more than two methylene-linked double bonds, showing that tri-unsaturated fatty acids are the in vivo source of up to 75% of MDA. The abundance of the combined pool of free and reversibly bound MDA did not change dramatically in stress, although a significant increase in the free MDA pool under oxidative conditions was observed. The half-life of infiltrated MDA indicated rapid metabolic turnover/sequestration. Exposure of plants to low levels of MDA using a recently developed protocol powerfully upregulated many genes on a cDNA microarray with a bias towards those implicated in abiotic/environmental stress (e.g. ROF1 and XERO2). Remarkably, and in contrast to the activities of other reactive electrophile species (i.e. small vinyl ketones), none of the pathogenesis-related (PR) genes tested responded to MDA. The use of structural mimics of MDA isomers suggested that the propensity of the molecule to act as a cross-linking/modifying reagent might contribute to the activation of gene expression. Changes in the concentration/localisation of unbound MDA in vivo could strongly affect stress-related transcription.

ARCHITECTING USER-CENTRIC PRIVACY-AS-A-SET-OF-SERVICES: DIGITAL IDENTITY RELATED PRIVACY FRAMEWORK

AbstractDigitalization gives to the Internet the power by allowing several virtual representations of reality, including that of identity. We leave an increasingly digital footprint in cyberspace and this situation puts our identity at high risks. Privacy is a right and fundamental social value that could play a key role as a medium to secure digital identities. Identity functionality is increasingly delivered as sets of services, rather than monolithic applications. So, an identity layer in which identity and privacy management services are loosely coupled, publicly hosted and available to on-demand calls could be more realistic and an acceptable situation. Identity and privacy should be interoperable and distributed through the adoption of service-orientation and implementation based on open standards (technical interoperability). Ihe objective of this project is to provide a way to implement interoperable user-centric digital identity-related privacy to respond to the need of distributed nature of federated identity systems. It is recognized that technical initiatives, emerging standards and protocols are not enough to guarantee resolution for the concerns surrounding a multi-facets and complex issue of identity and privacy. For this reason they should be apprehended within a global perspective through an integrated and a multidisciplinary approach. The approach dictates that privacy law, policies, regulations and technologies are to be crafted together from the start, rather than attaching it to digital identity after the fact. Thus, we draw Digital Identity-Related Privacy (DigldeRP) requirements from global, domestic and business-specific privacy policies. The requirements take shape of business interoperability. We suggest a layered implementation framework (DigldeRP framework) in accordance to model-driven architecture (MDA) approach that would help organizations' security team to turn business interoperability into technical interoperability in the form of a set of services that could accommodate Service-Oriented Architecture (SOA): Privacy-as-a-set-of- services (PaaSS) system. DigldeRP Framework will serve as a basis for vital understanding between business management and technical managers on digital identity related privacy initiatives. The layered DigldeRP framework presents five practical layers as an ordered sequence as a basis of DigldeRP project roadmap, however, in practice, there is an iterative process to assure that each layer supports effectively and enforces requirements of the adjacent ones. Each layer is composed by a set of blocks, which determine a roadmap that security team could follow to successfully implement PaaSS. Several blocks' descriptions are based on OMG SoaML modeling language and BPMN processes description. We identified, designed and implemented seven services that form PaaSS and described their consumption. PaaSS Java QEE project), WSDL, and XSD codes are given and explained.

Inducible malondialdehyde pools in zones of cell proliferation and developing tissues in Arabidopsis.

Malondialdehyde (MDA) is a natural and widespread genotoxin. Given its potentially deleterious effects, it is of interest to establish the identities of the cell types containing this aldehyde. We used in situ chemical trapping with 2-thiobarbituric acid and mass spectrometry with a deuterated standard to characterize MDA pools in the vegetative phase in Arabidopsis thaliana. In leaves, MDA occurred predominantly in the intracellular compartment of mesophyll cells and was enriched in chloroplasts where it was derived primarily from triunsaturated fatty acids (TFAs). High levels of MDA (most of which was unbound) were found within dividing cells in the root tip cell proliferation zone. The bulk of this MDA did not originate from TFAs. We confirmed the localization of MDA in transversal root sections. In addition to MDA in proliferating cells near the root tip we found evidence for the presence of MDA in pericyle cells. Remodeling of non-TFA-derived MDA pools occurred when seedlings were infected with the fungus Botrytis cinerea. Treatment of uninfected seedlings with mediators of plant stress responses (jasmonic acid or salicylic acid) increased seedling MDA levels over 20-fold. In summary, major pools of MDA are associated with cell division foci containing stem cells. The aldehyde is pathogen-inducible in these regions and its levels are increased by cellular mediators that impact defense and growth.

Metodologies de Desenvolupament Dirigides per Models

En aquest treball, s'introduiran dos de les metodologies de desenvolupament dirigides per models més significatives: Model Driven Architecture (MDA) i Domain Specific Modeling (DSM). Així mateix, es presentarà un estudi comparatiu d'algunes de les diferents eines existents actualment al mercat que els hi donen suport.

Generació automàtica de codi per aplicacions MVC amb AndroMDA i WebML

Aquest treball es centra es dues eines MDA: AndroMDA i WebML, amb les que es pretén generar una aplicació web basada en una arquitectura MVC. La introducció en el coneixement d'aquestes dues eines, es fa partint d'una visió global de l'arquitectura MDA, mostrant el seu funcionament, per a continuació passar a avaluar un conjunt d'eines MDA, de cara a realitzar una classificació en funció de les seves característiques.

Generació automàtica de codi Java amb ArgoUML

Aquesta memòria presenta un estudi de la generació automàtica de codi Java a partir de diagrames UML amb l'eina ArgoUML. El cicle de vida tradicional del programari presenta alguns problemes com la manca de sincronització entre codi i documentació, poca portabilitat i problemes de interoperabilidad. El paradigma MDA vol solucionar en part aquests problemes fent dels models el centre del desenvolupament del programari i apostant per la generació automàtica de models i codi.