applying-advanced-vitro-culturing-technology-to-study-human-gut

Applying Advanced In Vitro Culturing Technology to Study the Human Gut Microbiota

The human gut microbiota plays a vital role in both human health and disease. Studying the gut microbiota using an in vivo model, is difficult due to its ...

Agricultural Research Service, United States Department of Agriculture

Central to commercial fluid milk processing is the use of high temperature, short time (HTST) pasteurization to ensure the safety and quality of milk, and homogenization to prevent creaming of fat-containing milk. Ultra-high-temperature sterilization is also applied to milk and is typically used to extend the shelf life of refrigerated, specialty milk products or to provide shelf-stable milk. The structures of the milk proteins and lipids are affected by processing but little information is available on the effects of the individual processes or sequences of processes on digestibility. In this study, raw whole milk was subjected to homogenization, HTST pasteurization, and homogenization followed by HTST or UHT processing. Raw skim milk was subjected to the same heating regimens. In vitro gastrointestinal digestion using a fasting model was then used to detect the processing-induced changes in the proteins and lipids. Using sodium dodecyl sulfate-PAGE, gastric pepsin digestion of the milk samples showed rapid elimination of the casein and α-lactalbumin bands, persistence of the β-lactoglobulin bands, and appearance of casein and whey peptide bands. The bands for β-lactoglobulin were eliminated within the first 15min of intestinal pancreatin digestion. The remaining proteins and peptides of raw, HTST, and UHT skim samples were digested rapidly within the first 15min of intestinal digestion, but intestinal digestion of raw and HTST pasteurized whole milk showed some persistence of the peptides throughout digestion. The availability of more lipid droplets upon homogenization, with greater surface area available for interaction with the peptides, led to persistence of the smaller peptide bands and thus slower intestinal digestion when followed by HTST pasteurization but not by UHT processing, in which the denatured proteins may be more accessible to the digestive enzymes. Homogenization and heat processing also affected the ζ-potential and free fatty acid release during intestinal digestion. Stearic and oleic acids were broken down faster than other fatty acids due to their positions on the outside of the triglyceride molecule. Five different casein phosphopeptide sequences were observed after gastric digestion, and 31 sequences were found after intestinal digestion, with activities yet to be explored. Processing affects milk structure and thus digestion and is an important factor to consider in design of foods that affect health and nutrition. 

Effect of heat and homogenization on in vitro digestion of milk.

Apigenin is a major dietary flavonoid with many bioactivities, widely distributed in plants. Apigenin reaches the colon region intact and interacts there with the human gut microbiota, however there is little research on how apigenin affects the gut bacteria. This study investigated the effect of pure apigenin on human gut bacteria, at both the single strain and community levels. The effect of apigenin on the single gut bacteria strains , ,  GG, and , was examined by measuring their anaerobic growth profiles. The effect of apigenin on a gut microbiota community was studied by culturing a fecal inoculum under in vitro conditions simulating the human ascending colon. 16S rRNA gene sequencing and GC-MS analysis quantified changes in the community structure. Single molecule RNA sequencing was used to reveal the response of  to apigenin.  was effectively inhibited by apigenin when cultured alone, however, the genus  was enhanced when tested in a community setting. Single molecule RNA sequencing found that  responded to apigenin by up-regulating genes involved in DNA repair, stress response, cell wall synthesis, and protein folding. Taken together, these results demonstrate that apigenin affects both the growth and gene expression of .

Apigenin Impacts the Growth of the Gut Microbiota and Alters the Gene Expression of Enterococcus.

We report the successful preparation of reinforced electrospun nanofibers and fibrous mats of polyvinyl alcohol (PVA) via a simple and inexpensive method using stable tannic acid (TA) and ferric ion (Fe) assemblies formed by solution mixing and pH adjustment. Changes in solution pH change the number of TA galloyl groups attached to the Fe from one (pH < 2) to two (3 < pH < 6) to three (pH < 7.4) and affect the interactions between PVA and TA. At pH ~ 5.5, the morphology and fiber diameter size (FDS) examined by SEM are determinant for the mechanical properties of the fibrous mats and depend on the PVA content. At an optimal 8 wt % concentration, PVA becomes fully entangled and forms uniform nanofibers with smaller FDS ( < 0.05) and improved mechanical properties when compared to mats of PVA alone and of PVA with TA ( < 0.05). Changes in solution pH lead to beads formation, more irregular FDS and poorer mechanical properties ( < 0.05). The Fe inclusion does not alter the oxidation activity of TA ( > 0.05) suggesting the potential of TA-Fe assemblies to reinforce polymer nanofibers with high functionality for use in diverse applications including food, biomedical and pharmaceutical.

Peggy Tomasula

Dairy and Functional Foods Research Unit,
Eastern Regional Research Center,
Dairy and Functional Foods Research Unit, Eastern Regional Research Center

Agricultural Research Service, United States Department of Agriculture

Applying Advanced In Vitro Culturing Technology to Study the Human Gut Microbiota

Peggy Tomasula

Dairy and Functional Foods Research Unit,Eastern Regional Research Center,Dairy and Functional Foods Research Unit, Eastern Regional Research Center

Agricultural Research Service, United States Department of Agriculture

Applying Advanced In Vitro Culturing Technology to Study the Human Gut Microbiota

Dairy and Functional Foods Research Unit,
Eastern Regional Research Center,
Dairy and Functional Foods Research Unit, Eastern Regional Research Center