Protein extraction for proteome analysis from cacao leaves and meristems, organs infected by Moniliophthora perniciosa, the causal agent of the witches' broom disease

Preparation of high-quality proteins from cacao vegetative organs is difficult due to very high endogenous levels of polysaccharides and polyphenols. in order to establish a routine procedure for the application of proteomic and biochemical analysis to cacao tissues, three new protocols were developed; one for apoplastic washing fluid (AWF) extraction, and two for protein extraction - under denaturing and nondenaturing conditions. The first described method allows a quick and easy collection of AWF - using infiltration-centrifugation procedure - that is representative of its composition in intact leaves according to the smaller symplastic contamination detected by the use of the hexose phosphate isomerase marker. Protein extraction under denaturing conditions for 2-DE was remarkably improved by the combination of chemically and physically modified processes including phenol, SDS dense buffer and sonication steps. With this protocol, high-quality proteins from cacao leaves and meristems were isolated, and for the first time well-resolved 1-DE and 2-DE protein patterns of cacao vegetative organs are shown. It also appears that sonication associated with polysaccharide precipitation using tert-butanol was a crucial step for the nondenaturing protein extraction and subsequent enzymatic activity detection. It is expected that the protocols described here could help to develop high-level proteomic and biochemical studies in cacao also being applicable to other recalcitrant plant tissues.29112391240

Ecosystem approaches and health in Latin America Enfoques ecossistêmicos e saúde: vertentes e aplicações na América Latina

Important environmental changes that have become increasingly pronounced in the last two centuries and that are seriously affecting human health require the development of integrated and participatory scientific approaches that can result in proposals for institutional and public policy changes. The purpose of this article is to offer some elements that can contribute to a line of reflection based on studies with ecosystem approaches in the Latin America context. The authors begin with a brief description of current scientific literature in public health that links ecosystems and human health in Latin America; next, they describe and compare the two prevailing trends that form the basis for the theoretical and methodological debates on ecosystem approaches; they also review the empirical research in Latin America or concerning Latin American countries in which an ecosystem approach has been adopted. The results point to limited scientific output on the interface between ecosystems and human health; aspects involving public participation and implementation of institutional changes and public policies are still in a rather incipient stage.<br>As grandes mudanças ambientais que vêm se acentuando nos últimos dois séculos e que afetam a saúde humana exigem o desenvolvimento de abordagens científicas integradas, participativas e que resultem em proposições de mudanças institucionais e nas políticas públicas. O objetivo deste artigo é oferecer elementos para uma reflexão sobre uma linha destas abordagens integradas, os enfoques (eco)sistêmicos, na realidade latino-americana. Para alcançar este objetivo, realizamos breve descrição do quadro atual de produção científica no âmbito da Saúde Pública sobre ecossistemas e saúde humana na América Latina; descrevemos e comparamos as duas vertentes que se encontram na base do debate teórico e metodológico sobre os enfoques ecossistêmicos; analisamos os trabalhos empíricos que adotaram enfoques ecossistêmicos produzidos por pesquisadores da América Latina ou relacionados a países do continente. Os resultados apontam para uma pequena produção científica que considera a interface ecossistemas e saúde humana, sendo ainda bastante incipiente no que se refere aos aspectos referentes à participação e à formulação e implementação de mudanças institucionais e nas políticas públicas