These results confirm that bovine IgG is capable of surviving the passage through the entirety of the GI tract. Safety The safety population consisted of randomized subjects who received at least one dose of the investigational product. 20 g of SBI, with a significant difference between placebo and all doses of SBI (for 15 minutes to pellet remaining solids in the stool samples before harvesting the supernatant, which was stored at ?70C until shipped on dry ice for analysis. Bovine IgG in stool samples was quantified by a custom-developed ELISA utilizing a goat polyclonal antibody that specifically reacts to bovine IgG heavy BNP (1-32), human and light chains with minimum reactivity to human, mouse, and rat (Bethyl Laboratories, Inc.). Preliminary experiments with the custom assay established that the LOQ of bovine IgG in human stool homogenate was 2.7 pg/mg dry weight. Safety assessment The safety of SBI was evaluated in all subjects who consumed at least one packet of the investigational product. It was assessed by conducting physical examinations, evaluating vital signs, performing clinical laboratory testing, and monitoring AEs following oral administration of SBI. AEs were coded using the MedDRA coding dictionary and monitored for all subjects from the time of study initiation (informed consent) through the last administration of the investigational product. Therapy-emergent AEs (TEAE) were defined as any event with a start date occurring on or after first dose date of the investigational product or, if preexisting, worsening after first dose date of the investigational product. The study investigator was solely responsible for determining the relationship of an AE with the investigational product based on all the available information at the time of event recording, including any preexisting medical condition(s). A physical examination, including a comprehensive chemistry panel, complete blood count with differential, and urinalysis, was performed during the screening visit. Chemistry and hematology laboratory parameters were repeated at the end of the study visit (day 16). Symptom-directed physical examinations were performed at the study visits (ie, days 1, 2, 9, and 16) and for unscheduled clinic visits as needed. Subject-reported concomitant medications taken during the study were recorded at each visit. Statistical methods Following a single administration of SBI (5 g, 10 g, or 20 g) or placebo, plasma amino acid levels were quantitated as described earlier. Differences between the SBI dose groups and placebo were analyzed by comparing em C /em max and area under the curve (AUC) from 0 minutes to 180 BNP (1-32), human minutes (AUC0C180). The em C /em max and AUC were estimated using the post-administration amino acid responses over a 3-hour sampling interval and analyzed using a mixed model analysis of covariance (ANCOVA) technique appropriate for a two-period crossover design. A Rabbit Polyclonal to MED27 95% confidence interval and the associated em p /em -value for the least square (LS) means between the SBI groups (test) and placebo were provided for the amino acid response profiles. One subject from the 20 g group was excluded from the amino acid analysis due to protocol deviation. A sensitivity analysis was also conducted across participant response data within group, where data flagged as possible outliers were excluded from subsequent analyses. The method for outlier detection used a robust regression, leverage-point detection analysis, identifying outliers BNP (1-32), human as data points with a projected Mahalanobis distance 2. Differences in the mean change of stool bovine IgG concentrations between test doses of SBI (2.5 g BID; 5 g BID; 10 g BID) collected at day 9 and day 16 and corresponding baseline were analyzed using a mixed covariate-adjusted model for paired data approach. Baseline for all efficacy and safety variables was defined as the procedure performed during the screening visit or pre-dose on day 1, whichever provided the last pre-therapy assessment. All statistical analyses were performed using SAS? Version 9.3..