Mouse islets are commonly used in diabetes-related studies. Adequate amounts of good quality islets are prerequisites for a reliable investigation. We describe a protocol for islet isolation from mouse pancreas. Three major manipulations are employed in the islet isolation procedure: in situ pancreas perfusion with collagenase, pancreas digestion and islet purification. The whole procedure takes 30-45 min for each individual mouse. By using this protocol, a reasonable number of islets can be obtained in a relatively short period of time. This protocol has been proven to be practicable and reproducible. It can be easily followed by individuals who do not have previous experience in the related research field.
A simple and efficient microwave-assisted extraction of polyphenols from industrial apple pomace was developed and optimized by the maximization of the yield using response surface methodology. A Box-Behnken design was used to monitor the effect of microwave power, extraction time, ethanol concentration and ratio of solvent to raw material (g/mL) on the polyphenols yield. The results showed that the optimal conditions were as follows: microwave power 650.4 W, extraction time 53.7 s, ethanol concentration 62.1% and ratio of solvent to raw material 22.9:1. Validation tests indicated that the actual yield of polyphenols was 62.68±0.35 mg gallic acid equivalents per 100 g dry apple pomace with RSD=0.86% (n=5) under the optimal conditions, which was in good agreement with the predicted yield and higher than those of reflux and ultrasonic-assisted extraction methods. HPLC analysis indicated that the major polyphenols of apple pomace consisted of chlorogenic acid, caffeic acid, syrigin, procyanidin B2, (-)-epicatechin, cinnamic acid, coumaric acid, phlorizin and quercetin, of which procyanidin B2 had the highest content of 219.4 mg/kg.
The fermentation of cider by mixed cultures of Wickerhamomyces anomalus and Saccharomyces cerevisiae was carried out to study their effect on the cider quality. The results showed that growth of W. anomalus and S. cerevisiae was affected by each other during co-fermentation process. All the mixed cultures produced statistically the same level of ethanol as S. cerevisiae monoculture. The mixed fermentation could produce more variety and higher amounts of acetate esters, ethyl esters, higher alcohols, aldehydes, and ketones. Sensory evaluation demonstrated that ciders obtained from co-fermentation with W. anomalus gained higher scores than ciders fermented by pure S. cerevisiae, especially the co-fermentation cultures WS3, WS4, WS6, and WS8. Only 3 days of fermentation with W. anomalus in sequential mixtures were enough to improve the quality of cider. Wickerhamomyces anomalus could be used in association with S. cerevisiae to improve the quality of cider. The modulation of inoculation time may provide an effective means of manipulating cider aroma for different characteristics.
The contents of polyphenols and organic acids were affected by fermentation. Their changing profiles during fermentation process were dependent on the type of phenolic compounds and organic acids studied.
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