Grape seed is a potential source of edible protein, so effective extraction of protein component from grape seed seems to be very important. However, so far, such information is unavailable. In this study, the conditions for protein extraction from the grape seeds were optimized by the single factor test and the response surface methodology (RSM). A box-behnken design (BBD) was used for experimental design and analysis of the results to obtain the optimal extraction conditions. Solvent/meal ratio, extraction temperature, pH and extraction time were found to have a significant effect on the protein yield. From the single factor test, the optimal range of extraction conditions was obtained. Based on the RSM analysis, optimum extraction parameters were obtained as following: solvent/meal ratio 22.5/1 (v/w), extraction temperature 35℃, pH 9.8, and extraction time 29 min. Under the optimized conditions, the experimental values were in good agreement with those predicted by the model. These results help design the process of optimal protein extraction from grape seeds for future use in food industry.
The neuromuscular junction (NMJ) is a peripheral synaptic connection between presynaptic motor neurons and postsynaptic skeletal muscle fibers that enables muscle contraction and voluntary motor movement. Many traumatic, neurodegenerative, and neuroimmunological diseases are classically believed to mainly affect either the neuronal or the muscle side of the NMJ, and treatment options are lacking. Recent advances in novel techniques have helped develop in vitro physiological and pathophysiological models of the NMJ as well as enable precise control and evaluation of its functions. This paper reviews the recent developments in in vitro NMJ models with 2D or 3D cultures, from organ‐on‐a‐chip and organoids to biohybrid robotics. Related derivative techniques are introduced for functional analysis of the NMJ, such as the patch‐clamp technique, microelectrode arrays, calcium imaging, and stimulus methods, particularly optogenetic‐mediated light stimulation, microelectrode‐mediated electrical stimulation, and biochemical stimulation. Finally, the applications of the in vitro NMJ models as disease models or for drug screening related to suitable neuromuscular diseases are summarized and their future development trends and challenges are discussed.
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