Obtenção de microcápsulas do éster etílico do ácido docosa-hexaenoico (DHAEE) por atomização: otimização através de planejamento experimental

Docosahexaenoic acid is an essential polyunsaturated fatty acid with important metabolic activities. Its conjugated double bonds make it susceptible to decomposition. Its stability may be improved through fatty acid entrapment with a spray-drying technique; however, the many parameters involved in this technique must be considered to avoid affecting the final product quality. Therefore, this study aimed to evaluate the entrapment conditions and yields of fish oil enriched with docosahexaenoic acid ethyl ester. Microcapsules were obtained from Acacia gum using a spray-drying technique. The experimental samples were analyzed by chromatography and delineated by Statistica software, which found the following optimum entrapment conditions: an inlet temperature of 188 °C; 30% core material; an N2 flow rate of 55 mm; and a pump flow rate of 12.5 mL/minute. These conditions provided a 66% yield of docosahexaenoic acid ethyl ester in the oil, corresponding to 19.8% of entrapped docosahexaenoic acid ethyl ester (w/w). This result was considered significant since 30% corresponded to wall material.O ácido docoso-hexaenoico é um ácido graxo poli-insaturado essencial que desempenha importantes ações metabólicas. Entretanto, por possuir duplas ligações conjugadas torna-se suscetível à decomposição. Uma das formas de minimizar esta possível decomposição é o emprego da técnica de atomização para microencapsulação. Porém, esta técnica envolve uma série de parâmetros de processo, que podem vir a alterar a qualidade do produto final. Assim, o objetivo deste trabalho foi microencapsular óleo de peixe enriquecido no éster etílico do ácido docoso-hexaenoico (DHAEE-85%), variando condições operacionais e avaliar o rendimento pela análise por cromatografia gasosa, após extração das microcápsulas. Para tanto, foi utilizado o processo de microencapsulação por atomização e o agente encapsulante foi a goma arábica. A avaliação cromatográfica de vários experimentos delineados pelo software Statistica, mostrou que os pontos ótimos para obtenção das microcápsulas de DHAEE foram: temperatura de entrada 188 °C, porcentagem de recheio 30%, vazão de nitrogênio 55 mm N2 e vazão da bomba de 12,5 mL/minuto. Estas condições de processo foram testadas experimentalmente, resultando no teor de 66% m/m de DHAEE no óleo extraído, valor correspondente a 19,8% m/m de DHAEE encapsulado, valor considerado satisfatório uma vez que 30% das microcápsulas correspondiam teoricamente ao material de parede.589596Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field