Carbohydrate and mineral removal during the production of low-phytate soy protein isolate by combined electroacidification and high shear tangential flow ultrafiltration.

In this work, soy protein isolates were produced by a combination of electroacidification and high shear tangential flow hollow fiber ultrafiltration with a 100 kDa membrane under constant pressure. The filtration performance was evaluated by comparing the filtration time and the final product composition for an electroacidified (pH 6) and a non-electroacidified (pH 9) soy protein extract. The removal of carbohydrates during the filtration was always consistent with the theoretical predictions (based on free permeability assumption) for both the electroacidified and the non-electroacidified feeds. A higher removal of calcium, magnesium, and phytic acid was achieved during the filtration of the electroacidified feed compared to the non-electroacidified feed. However, the electroacidification pretreatment had a negative impact on the permeate flux and resulted in more significant membrane fouling with correspondingly longer filtration times. A discontinuous diafiltration enhanced the removal of carbohydrates and minerals, thus yielding a product with higher protein content but was unable to improve the permeate flux for the electroacidified feed.

Design and analysis of a long-term live-cell imaging chamber for tracking cellular dynamics within cultured human islets of Langerhans.

A means of expanding islet cell mass is urgently needed to supplement the limited availability of donor islets of Langerhans for transplant. Live cell imaging of human islets in culture has the potential to identify the specific cells and processes involved in islet expansion. A novel imaging chamber was developed to facilitate long-term three-dimensional imaging of human islets during transformation. Islets have been induced to transform into duct-like epithelial cystic structures and revert back to glucose responsive endocrine cells under appropriate conditions (Jamal et al. Cell Death Differ. 2005 12:702-712). Here we aim to further our understanding by characterizing the process at a single cell level over time-essentially constructing a high resolution recorded history of each cell and its progeny during transformation and reversion. The imaging chamber enables high resolution imaging of three-dimensional islets while maintaining the structure of the islet cells and intercellular matrix components. A mathematical model was developed to validate the imaging chamber design by determining the required chamber dimensions to avoid introduction of oxygen and nutrient transport limitations. Human islets were embedded in collagen in the imaging chamber and differential interference contrast time course images were obtained at 3 min intervals. Immunofluorescent imaging confirmed that islet phenotype was maintained for at least 5 days during imaging. Analysis of the time courses confirms our ability to identify and track individual cells over time and to observe cell death and phenotype transformation in isolated human islets.