Conclusions The present findings indicate that unknown metabolites produced by probiotic Lactobacilli elicit rapid, non-genomic responses in the ability of intestinal epithelial cells to transport glucose. Whether genomic responses are also induced is unknown. The responses of Ca and Na uptake to bacterial metabolites (18,34) suggest the rapid stimulation of glucose transport triggered by the metabolites from Lactobacilli will be shared by carriers for other nutrients. There is an obvious need to identify the specific bacterial metabolites that elicit desired responses (i.e., increased nutrient absorption,

immunomodulation, etc) and the bacterial species and conditions BYL719 cost that promote the production. Methods Probiotic Bacteria Culture A working culture of L. acidophilus (ATCC#4356) was propagated for 48 h at 37°C in DeMan, Rogosa and Sharpe (MRS) broth (Difco, Becton-Dickinson, Franklin Lakes, NJ) in a continuous shaker placed inside an anaerobic chamber with an atmosphere of 80% nitrogen, 10% carbon

dioxide, and 10% hydrogen. The bacterial cells were sedimented by centrifugation (519 × g; 5 minutes) and were washed twice with sterilized water. The cells were suspended in a solution of 80% Dulbecco’s Phosphate-Buffered Saline and 20% glycerol, and stored at -80°-C until MM-102 in vitro used for experiments. After characterizing a response of Caco-2 cells to the supernatant after culture of L. acidophilus, MK-0457 ic50 additional strains of Lactobacilli were obtained from Wyeth Nutrition (Collegeville, PA 19426, USA) for comparative purposes and working cultures were similarly prepared. These included L. amylovorus (ATCC#33620), L. gallinarum (ATCC#33199), L. gasseri (ATCC#33323), and L. johnsonii (ATCC#33200). Chemically Defined Media The probiotic bacteria were cultured anaerobically

to mimic conditions in the colon using a chemically defined medium (CDM; Table 1) [34] that was prepared without Dolutegravir carbohydrate (pH = 6.5; 400 mOsm), filter sterilized (0.20 μm, Millipore, Billerica, MA), and stored at 4°C until used. A preliminary trial identified carbohydrates that would support the growth of L. acidophilus by adding arabinose, fructose, glucose, mannose, ribose, and xylose to the CDM at a concentration of 110 mM. Growth of L. acidophilus in MRS broth, which has 110 mM glucose, was used as a positive control. The CDM with different sources of carbohydrates and the MRS were pre-reduced and made anaerobic by placing them in the anaerobic chamber for 12-18 h before they were inoculated with the L. acidophilus suspension (200 μL with 109 CFU/ml in 500 ml). Aliquots were removed immediately after the inoculation and every 4 h thereafter during 80 h of anaerobic growth at 37°C and optical density at 600 nm was recorded to track bacterial growth and to define three different phases of the growth curves; the lag phase before rapid growth, at the middle of exponential growth, and after the start of the stationary phase.