Adipose-derived stromal cells (ASCs) are usually isolated by digestion with collagenase. We have compared alternative methods to isolate ASCs in a more economically viable protocol. Nine protocols using red blood cells lysis buffer solution, trypsin, collagenase and centrifugation were compared; the isolation rate, cell viability, expansion rate, immunophenotype and differentiation in adipogenic and osteogenic lineages were analyzed. ASCs were isolated and successfully maintained by digestion with trypsin. Cells presented similar immunophenotypes, adipogenic differentiation and in vitro proliferation but an osteogenic differentiation capacity up to seven times higher than ASCs isolated by collagenase. This alternative protocol is thus efficient and more cost-effective than the commonly-used methods and may represent a promising protocol for obtaining ASCs for bone tissue engineering.

Introduction Apoptosis of neutrophils (polymorphonuclear neutrophils [PMNs]) may limit inflammatory injury in sepsis and acute respiratory distress syndrome (ARDS), but the relationship between the severity of sepsis and extent of PMN apoptosis and the effect of superimposed ARDS is unknown. The objective of this study was to correlate neutrophil apoptosis with the severity of sepsis and sepsis-induced ARDS.

RESUMOMuitos indivíduos deixam de consumir leite pelo fato de haver alguma restrição, normalmente, por dois motivos: intolerância à proteína ou intolerância à lactose. Esta última atinge quase a metade da população mundial. O objetivo deste trabalho foi avaliar um processo híbrido combinando ultrafiltração (UF), nanofiltração (NF) e hidrólise enzimática para a obtenção de leite pasteurizado com reduzido teor de lactose. Leite desnatado foi ultrafiltrado em unidade piloto de filtração por membranas, separando duas correntes: permeado e retido. A corrente do permeado da ultrafiltração foi coletada e armazenada sob refrigeração para posterior utilização no processo de nanofiltração. A corrente do permeado da NF foi, então, reincorporada na do retido da UF, na qual estavam presentes proteínas, gordura, sais minerais e vitaminas do leite. No produto resultante, foi realizada a hidrólise enzimática com o uso da enzima betagalactosidade. O produto final foi pasteurizado a 75 ºC por 15 s. Análises físico-químicas e microbiológicas foram realizadas para caracterização do produto, tendo tais parâmetros sido definidos com base na legislação e em outros estudos científicos. O produto final apresentou-se dentro dos parâmetros exigidos pela legislação, tendo a redução de lactose chegado a 93% em relação ao teor inicial, representando uma boa alternativa para as indústrias de laticínios, agregando valor ao produto e dispondo um alimento alternativo para os consumidores com intolerância à lactose. Palavras-chave: Separação por membranas. Lactose. Hidrólise enzimática. ABSTRACTMany individuals fail to consume milk because there is some restriction, usually for two reasons: protein intolerance or lactose intolerance. The latter reaches almost half the world's population. The aim of this study was to evaluate a hybrid process combining ultrafiltration (UF), nanofiltration (NF) and enzymatic hydrolysis to obtain pasteurized milk with reduced lactose content. Skim milk was ultrafiltered in a pilot unit membrane filtration, separating two streams: penetration and retention. The current ultrafiltration permeate was collected and stored under refrigeration for later use in nanofiltration process. The NF permeate stream was then remixed with the UF retentate, which were present in the protein, fat, minerals and vitamins of milk. In the resulting product, enzymatic hydrolysis was carried out with the use of the enzyme betagalactosidade. The final product was pasteurized at 75 °C for 15 sec. Physical-chemical and microbiological analyzes were performed for characterization of the product, and such parameters have been defined based on legislation and other scientific studies. The final product was within the parameters required by the legislation, and the reduction of lactose reached 93% compared to the initial rate, representing a good alternative to dairy industries, adding value to the product and providing a food alternative for consumers with lactose intolerance.

To disclose the mechanisms surrounding obesity, we selected microRNAs (miRNAs) that target genes involved in adipogenesis, angiogenesis, and inflammation and compared their expression levels in the subcutaneous adipose tissue of 40 obese and nonobese women. Mature miRNAs were extracted from subcutaneous adipose tissue samples that were collected during surgery and quantified by real-time polymerase chain reaction. miR-16 was overexpressed in the nonobese group (n-expression ratio = − 151.1; P < 0.001). Furthermore, the expression levels of two other miRNAs were significantly correlated with waist circumference in nonobese women (miR-27b, r = 0.453; P = 0.027 and miR-424-5p, r = 0.502, P = 0.014). Central and total subcutaneous adipose tissue thicknesses were correlated with miR-424-5p levels (r = 0.506, P = 0.034 and r = 0.475, P = 0.046, respectively) in the nonobese group. In the obese group, miR-424-5p expression was correlated with body mass index (r = 0.582, P = 0.018). miR-16 and miR-424 have shown correlations with body-fat-mass-related parameters. Because these miRNAs have vascular endothelial growth factor (VEGF) and its receptors as target genes, they may be involved in the alterations of angiogenesis observed in obesity. In addition, higher levels of miR-27 and miR-424 were correlated with higher fat depot measurements in nonobese women. These results highlight the importance of miRNA expression in subcutaneous adipose tissue and encourage further investigation of miRNAs as prognostic markers.

Fouling in whey ultrafiltration is still a problem to be solved and modelling the permeate flux is still a challenge. A set of experiments of whey ultrafiltration was performed and it was demonstrated that the cake filtration model accurately represents the permeate flux over time despite exist more complex models to describe the phenomenon. In less than 10 s an irreversible resistance is established, and the cake starts to build up. It was observed changes in protein rejection in a range of 98 and 94% when a higher cake layer was set up, but it was not noticed strong evidences that the layer deposited on the membrane surface drives this change.Formação de torta e redução de desempenho da ultrafiltração de soro de leite RESUMO. A colmatação da membrana durante a ultrafiltração de soro de leite ainda é um problema a ser resolvido e a modelagem do fluxo permeado permanece como um desafio. Uma série de experimentos de ultrafiltração de soro foi realizada e foi demonstrado que o modelo de torta de filtração pode representar o fluxo permeado com precisão, apesar de existirem modelos mais complexos para descrever o fenômeno. Em menos de 10 s, uma resistência irreversível é estabelecida e a torta começa a se formar. Foram observadas variações na rejeição de proteína de 98 a 94% quando uma espessa camada de torta foi depositada, mas não foram observadas fortes evidências de que essa camada de torta é a causa dessa variação na rejeição de proteína.Palavras-chave: modelagem de fluxo, membrana, laticínios, bloqueio de poros, rejeição da membrana. 518Brião et al.