Individuals with inflammatory bowel diseases experience an elevated risk of colorectal cancer driven by chronic inflammation. Current systemic immunosuppressive therapies often cause severe side effects. Live oral biotherapeutics are an emerging treatment modality that directly target the intestines. We have engineered a probiotic Saccharomyces boulardii strain that expresses targeting ligands to bind fibronectin on inflamed mucosa and secretes anti-tumor necrosis factor nanobodies locally to reduce inflammation. We previously demonstrated that engineering S. boulardii to bind fibronectin enhanced colonization and reduced inflammation in a DSS colitis model. Here, we tested the anti-cancer potential of engineered S. boulardii using a well-established model of IBD-associated CRC, azoxymethane-treated interleukin 10-deficient (AOM/Il10−/−) mice. These mice develop inflammation and invasive tumors that model those found in inflammatory bowel disease. Mice were orally administered engineered S. boulardii at two dosing frequencies, unmodified S. boulardii, or placebo throughout the 18-week model. Colons were harvested for gross, histological, and molecular evaluation for inflammation and tumorigenesis. Histological colon inflammation was reduced by twice weekly dosing of engineered and unmodified S. boulardii. Engineered S. boulardii reduced gross tumor number in a dose-dependent manner, with median tumor counts reduced from 7.5 to 2 per mouse (p < 0.0002 vs. placebo). Unmodified S. boulardii similarly reduced gross tumor number. Colonization studies revealed that engineered S. boulardii failed to colonize for greater time or density vs. unmodified S. boulardii. Together our data indicate that engineering S. boulardii does not reduce its ability to decrease inflammation-associated tumorigenesis, and that further host-binding target optimization is required to enhance colonization and anti-cancer effects.

This randomized, placebo-controlled, double-blind, parallel trial investigated whether generally healthy adult, nonelite runners would have a greater time-to-exhaustion during submaximal treadmill running with probiotic versus placebo supplementation. It was hypothesized that the probiotic would impact training progression by reducing gastrointestinal (GI) and cold/flu symptoms. Participants who typically ran ≥24 km/week, ran or cross-trained 3-5 days per week, and had a maximal oxygen intake (V̇O2 max) in the 60-85th percentile were enrolled. V̇O2 max was used to establish individualized workload settings (85% of V̇O2 max) for the submaximal endurance tests at baseline and following 6 weeks of supplementation with a probiotic (Lactobacillus helveticus Lafti L10, 5×109 CFU/capsule/day) or placebo. Participants self-reported GI and cold/flu symptoms and physical activity via daily and weekly questionnaires. Outcomes were tested using a linear model to determine if mean response values adjusted for baseline differed between groups. Twenty-eight participants (n = 14/group), aged 25 ± 5 years (mean ± SD) with a body mass index of 23 ± 3 kg/m2, completed the study. At the final visit the probiotic group had a lower time-to-exhaustion versus the placebo group (P = 0.01) due to an increase in time-to-exhaustion with the placebo (1344 ± 188 to 1565 ± 219 s, P = 0.01) with no change with the probiotic (1655 ± 230 to 1547 ± 215 s, P = 0.23). During the intervention, the probiotic group completed fewer aerobic training sessions per week (P = 0.02) and trained at a lower intensity (P = 0.007) versus the placebo group. Few GI and cold/flu symptoms were reported with no differences between groups. Time-to-exhaustion increased in the placebo group, possibly due to differences in training habits.

BACKGROUND The American Association for the Study of Liver Disease recommends screening patients with cirrhosis for hepatocellular carcinoma (HCC) using imaging with or without alpha-fetoprotein every six months. Unfortunately, screening rates remain inadequate. AIM To assess root causes of screening failure in a subspecialty hepatology clinic. METHODS The authors identified patients with cirrhosis seen in a subspecialty hepatology clinic and determined whether they underwent appropriate screening, defined as two cross-sectional images between five and seven months apart. The authors characterized the primary driver of screening failure. Finally, other hepatologists were surveyed to determine provider perceptions of screening failure causes. RESULTS 1034 patients were identified with an average age of 61 years and a mean MELD of 8.1 ± 3.8. Hepatitis C virus was the most common cirrhosis etiology. 489 (47%) underwent appropriate screening. No demographic or clinical differences were detected between those who underwent appropriate screening and those who did not. The most common etiologies of screening failure, in descending order, were: radiology unable to schedule timely imaging, provider did not order imaging, patient canceled follow up appointment, appointments scheduled too far apart, lost to follow up, no-show to radiology appointment, and provider canceled appointment. Hepatologists surveyed believed the most common cause of screening failure was no-show to radiology. CONCLUSION Rates of screening were poor even in a subspecialty hepatology clinic. Screening failure was mostly due to systemic factors such as radiology availability and time between hepatology appointments rather than individual error.

Kefir, a fermented beverage made from kefir grains, has gained immense popularity around the world due to its potential health-promoting properties. Kefir beverages are both marketed commercially and brewed privately by individuals. Both milk and sugar solutions can be used as substrates with various additives included based on consumer preference. Fermentation occurs via microorganisms including lactic acid bacteria, acetic acid bacteria, and yeasts, which are naturally present in kefir grains. Health-promoting effects of kefir are thought to occur through immune, gastrointestinal, and metabolic regulation. Both clinical trials and mechanistic studies in cell culture and animal models have explored these effects. Studies in vitro and in animals have shown the ability of kefir and kefir components to antagonize pathogens, reduce proinflammatory cytokine production, contribute to cytotoxicity of tumor cell lines and reduce tumor burden, and improve serum glycemic and lipid profiles. However, some data from clinical trials are conflicting, and the precise mechanisms by which kefir promotes well-being are not completely defined. This review summarizes the current body of evidence in both cell culture and animal models that provide insight into the mechanisms by which kefir beverages may protect consumers from enteric infections and improve immune and metabolic health. We believe that readers will gain knowledge helpful for both developing more targeted mechanistic studies and selecting informative outcomes when designing clinical studies.

Background Patients require vitamin and mineral supplementation after bariatric surgery to prevent the development of micronutrient deficiencies. Consuming oral supplements is challenging due to gastric volume restriction. A transdermal patch dosage form may provide adequate micronutrient supplementation without pill burden. The study aims to determine the percentage of patients who have two or more nutritional deficiencies one year after surgery and to determine serum nutrient concentrations and gastrointestinal symptoms over time. Methods Patients who planned to undergo bariatric surgery and preferred transdermal patches versus oral supplements were recruited during preoperative office visits. Enrolled patients were instructed to use a transdermal multivitamin patch as per the manufacturer's instructions. Serum nutrient concentrations and Gastrointestinal Symptom Response Scale scores were determined at baseline and three months, six months, and one year after surgery. Results Ninety-two participants completed the study protocol. Twenty-five participants had a full panel of study labs one year after surgery. Among these patients, 19% had two or more micronutrient deficiencies. Vitamin D was the most common deficiency followed by vitamin B6; however, median serum concentrations of both nutrients increased over time. Vitamin B1, folate, and zinc deficiencies were also observed. There were no changes in gastrointestinal symptoms. Conclusions Additional studies, including randomized controlled trials, are required to determine if the PatchMD Multivitamin Plus patch (Pilot Rd. STE. B, Las Vegas) can provide adequate supplementation of vitamins and minerals. The patch was not associated with changes in gastrointestinal symptoms.

Cardiovascular disease (CVD) prevalence remains elevated globally. We have previously shown that a one-week lifestyle "immersion program" leads to clinical improvements and sustained improvements in quality of life in moderate to high atherosclerotic CVD (ASCVD) risk individuals. In a subsequent year of this similarly modeled immersion program, we again collected markers of cardiovascular health and, additionally, evaluated intestinal microbiome composition. ASCVD risk volunteers (n = 73) completed the one-week "immersion program" involving nutrition (100% plant-based foods), stress management education, and exercise. Anthropometric measurements and CVD risk factors were compared at baseline and post intervention. A subgroup (n = 22) provided stool, which we analyzed with 16S rRNA sequencing. We assessed abundance changes within-person, correlated the abundance shifts with clinical changes, and inferred functional pathways using PICRUSt. Reductions in blood pressure, total cholesterol, and triglycerides, were observed without reduction in weight. Significant increases in butyrate producers were detected, including Lachnospiraceae and Oscillospirales. Within-person, significant shifts in relative abundance (RA) occurred, e.g., increased Lachnospiraceae (+58.8% RA, p = 0.0002), Ruminococcaceae (+82.1%, p = 0.0003), Faecalibacterium prausnitzii (+54.5%, p = 0.002), and diversification and richness. Microbiota changes significantly correlated with body mass index (BMI), blood pressure (BP), cholesterol, high-sensitivity C-reactive protein (hsCRP), glucose, and trimethylamine N-oxide (TMAO) changes. Pairwise decreases were inferred in microbial genes corresponding to cancer, metabolic disease, and amino acid metabolism. This brief lifestyle-based intervention improved lipids and BP and enhanced known butyrate producers, without significant weight loss. These results demonstrate a promising non-pharmacological preventative strategy for improving cardiovascular health.

Wilson's disease is a complex multi-organ disease characterized by impaired biliary excretion of copper and resulting deposition of excess copper in the liver and other organs. It has a wide range of clinical presentations, and diagnosis often requires a high degree of clinical suspicion, especially in patients with multiple other comorbidities. We present the case of a 37-year-old woman with a complex medical and psychiatric history who was admitted for chronic diarrhea, hepatic enzyme elevation, electrolyte abnormalities, hyperammonemia, and methicillin-sensitive Staphylococcus aureus bacteremia. She was eventually found to have low serum ceruloplasmin level and elevated urine copper levels. Though confirmatory liver biopsy was not performed due to bowel wall rupture and septic shock, most of her symptoms and lab abnormalities could be explained by an underlying diagnosis of Wilson's disease. We present this case primarily as a cautionary tale. This patient was not lacking in medical attention prior to this prolonged hospitalization; however, her psychiatric issues and fibromyalgia management were the predominant foci during her frequent primary care office visits and likely distracted from the patient's chronic laboratory abnormalities. More vigilant laboratory evaluation of underlying medical conditions in psychiatric patients may be warranted in order to prevent serious complications of such conditions.

Microbial metabolism in the gut may alter human bile acid metabolism in a way that beneficially affects lipid homeostasis and therefore cardiovascular disease risk. Deconjugation of bile acids by microbes is thought to be key to this mechanism but has yet to be characterised in blood and stool while observing lipid markers. The aim of this study was to determine the effect of 3 different probiotic strains on plasma and stool bile acids in the context of lipid and glucose metabolism. In this 18-week, randomised, double-blind crossover study, healthy adults (53±8 years) with a high waist circumference underwent a 1-week pre-baseline period and were then randomised to receive 1 capsule/day of Bacillus subtilis R0179 (2.5×109 cfu/capsule; n=39), Lactobacillus plantarum HA-119 (5×109 cfu/capsule; n=38), Bifidobacterium animalis subsp. lactis B94 (5×109 cfu/capsule; n=37) or placebo for 6 weeks. Following a 3-week washout and second pre-baseline week, participants were crossed to the other intervention for 6 weeks followed by a 1-week post-intervention period. Blood and stool samples were collected at the beginning and end of each intervention to measure bile acids, serum lipid profiles, and glucose and insulin levels. Data from the placebo intervention were combined for all participants for analyses. In obese participants, the difference (final-baseline) in the sum of deconjugated plasma bile acids was greater with consumption of B. subtilis (691±378 nmol/l, P=0.01) and B. lactis (380±165 nmol/l, P=0.04) than with placebo (98±176 nmol/l, n=57). No significant differences were observed for any probiotics for stool bile acids, serum lipids, blood glucose or insulin. These data suggest that B. subtilis and B. lactis had no effect on glucose metabolism or serum cholesterol but increased deconjugated plasma bile acids in obese individuals. Additional studies should be conducted to confirm these findings and explore potential mechanisms. This trial was registered at clinicaltrials.gov as NCT01879098.

Background: Rhinoconjunctivitis-specific quality of life is often reduced during seasonal allergies. The Mini Rhinoconjunctivitis Quality of Life Questionnaire (MRQLQ) is a validated tool used to measure quality of life in people experiencing allergies (0 = not troubled to 6 = extremely troubled). Probiotics may improve quality of life during allergy season by increasing the percentage of regulatory T cells (Tregs) and inducing tolerance.Objective: The objective of this study was to determine whether consuming Lactobacillus gasseri KS-13, Bifidobacterium bifidum G9-1, and B. longum MM-2 compared with placebo would result in beneficial effects on MRQLQ scores throughout allergy season in individuals who typically experience seasonal allergies. Secondary outcomes included changes in immune markers as part of a potential mechanism for changes in MRQLQ scores.Design: In this double-blind, placebo-controlled, parallel, randomized clinical trial, 173 participants (mean ± SEM: age 27 ± 1 y) who self-identified as having seasonal allergies received either a probiotic (2 capsules/d, 1.5 billion colony-forming units/capsule) or placebo during spring allergy season for 8 wk. MRQLQ scores were collected weekly throughout the study. Fasting blood samples were taken from a subgroup (placebo, n = 37; probiotic, n = 35) at baseline and week 6 (predicted peak of pollen) to determine serum immunoglobulin (Ig) E concentrations and Treg percentages.Results: The probiotic group reported an improvement in the MRQLQ global score from baseline to pollen peak (-0.68 ± 0.13) when compared with the placebo group (-0.19 ± 0.14; P = 0.0092). Both serum total IgE and the percentage of Tregs increased from baseline to week 6, but changes were not different between groups.Conclusions: This combination probiotic improved rhinoconjunctivitis-specific quality of life during allergy season for healthy individuals with self-reported seasonal allergies; however, the associated mechanism is still unclear. This trial was registered at clinicaltrials.gov as NCT02349711.

BackgroundQuality of life (QOL) is often reduced during seasonal allergies due to troubles from nose and eye symptoms. Probiotics modulate immune function and may improve QOL for some individuals during allergy season. Induction of tolerance by regulatory T cells (Tregs) may be involved in this processMethodsIn this double‐blind, placebo‐controlled, randomized clinical trial, 173 participants (26.0±1.2 y [mean±SEM] and 62.8% females, probiotic group; 27.6±1.4 y and 74.7% females, placebo group) who self‐identified as having seasonal allergies received either a probiotic (2 capsules/day, 1.5 billion cells/capsule) or placebo capsule during spring allergy season for 6 weeks. QOL was assessed throughout the study using the Mini Rhinoconjunctivitis Quality of Life Questionnaire (MRQLQ), which reports the average score for 14 symptoms rated from 0 (not troubled) to six (extremely troubled). Intestinal microbial profiles were determined via 16S rRNA sequencing from stool samples collected at baseline and at final (week 6). Fasting blood samples were taken from a subgroup (n=72) at baseline and final to determine immune parameters; serum total IgE and Tregs were quantified via ELISA and flow cytometry, respectively.ResultsTotal IgE increased from baseline to final regardless of group (data were log transformed for analysis; untransformed: 547±62.5 vs. 595±66.6 ng/mL, p=0.033), indicating a sustained response to allergens. Both groups reported decreases in MRQLQ scores from baseline to final, indicating a possible placebo effect or induction of tolerance; however, the negative change in global MRQLQ scores from baseline to final was significantly larger in magnitude in the probiotic group (−0.66±0.13 vs. −0.23±0.16, p=0.039), indicating higher QOL. Several domain scores from the MRQLQ also differed significantly between groups (−0.55±0.15 vs. −0.05±0.16, p=0.017 for activities; −0.71±0.18 vs. −0.09±0.19, p=0.016 for nose symptoms; −0.90±0.18 vs. −0.37±0.18, p=0.008 for other symptoms). Alpha and beta diversity indices to measure representative intestinal bacterial phyla did not differ between groups. The percentage of Tregs increased from baseline to final in all participants, but changes were not different between groups.ConclusionsThis combination of probiotics resulted in a higher self‐reported QOL during allergy season for healthy individuals who experience seasonal allergies. However, further research is needed to elucidate the mechanism by which probiotics influence allergy‐related QOL and to establish the clinical relevance for the change in the MRQLQ scores.Support or Funding InformationWakunaga of America Co., Ltd.

BackgroundProbiotics have been shown to transiently modify the intestinal microbiota by favoring the growth of endogenous bacteria possessing bile salt hydrolase (BSH) activity. The BSH activity promotes deconjugation and excretion of bile acids and shifts metabolism from cholesterol synthesis to bile formation, thereby attenuating systemic cholesterol levels and decreasing risk of cardiovascular diseases.ObjectiveThe aim of this study was to determine the effect of 3 different probiotic strains on bile acid and glucose metabolism.MethodsIn a randomized, double‐blind crossover study, healthy middle‐aged adults (52.6 ± 7.7 years old) with a larger waist circumference (>102 cm and >88 cm for males and females, respectively) received 1 capsule/day of Bacillus subtilis R0179 (2.5 × 109 CFU/capsule) (n=39), Lactobacillus plantarum HA 119 (5 × 109 CFU/capsule) (n=38), Bifidobacterium animalis subsp. lactis B94 (5 × 109 CFU/capsule) (n=37) or placebo for 6 wks. Following a 4‐wk washout period, participants were crossed to the other treatment (i.e. probiotic to placebo or placebo to probiotic). Stool frequency and consistency were recorded on a daily basis. Blood and stool samples were collected at the beginning and end of each intervention to measure bile acids. Serum lipids and markers of glucose metabolism were measured in the blood. Weekly measures included gastrointestinal symptoms (Gastrointestinal Symptom Rating Scale (GSRS)) and physical activity levels (Internal Physical Activity Questionnaire (IPAQ)).ResultsThe change in deconjugated plasma bile acids was greater with consumption of B. subtilis R0179 (691±378nmol/L, p=0.01) and B. animalis subsp. lactis B94 (380±165nmol/L, p=0.04) than with combined placebo data (98±176nmol/L). This change was only observed in obese participants (Body Mass Index ≥ 30). Additionally, there was a trend towards an increase in HDL plasma levels during the L. plantarum HA 119 intervention (3.0±1.3mg/dL, p=0.0774) versus the placebo (−0.3±0.8mg/dL). No significant differences were observed for any probiotics for stool bile acids, other serum lipids, gastrointestinal symptoms and function, blood glucose and insulin, and physical activity levels.ConclusionsThese data suggest that B. subtilis R0179 and B. animalis subsp. lactis B94 had no effect on glucose metabolism or serum cholesterol but significantly increased deconjugated plasma bile acids. It is possible that de novo bile acid synthesis was inhibited by these probiotics through the Farnesoid X Receptor (FXR) in the ileum. Additional clinical studies should be conducted to confirm these findings and explore the mechanism by which probiotics may simultaneously modulate cholesterol absorption and catabolism to bile acid.Support or Funding InformationLallemand Health Solutions

Psychological stress is associated with gastrointestinal (GI) distress. This secondary analysis from a randomised, double-blind, placebo-controlled study examined whether three different probiotics could normalise self-reported stress-associated GI discomfort and reduce overall self-reported stress. Undergraduate students (n=581) received Lactobacillus helveticus R0052, Bifidobacterium longum ssp. infantis R0033, Bifidobacterium bifidum R0071, or placebo. Participants self-reported 2 outcomes for a 6-week period, which included final academic exams: daily level of stress (0=no stress to 10=extremely stressed) and weekly three diarrhoea-related symptoms (DS, 1=no discomfort to 7=severe discomfort) using the GI Symptom Rating Scale. Self-reported stress was positively related to DS (P=0.0068). Mean DS scores were lower with B. bifidum versus placebo at week 2 at the average level of stress and the average body mass index (BMI). DS scores were lower with B. bifidum at week 5 versus week 0 and 1 and with B. infantis R0033 at week 6 versus week 0. DS scores were higher when antibiotics were used in the prior week with placebo (P=0.0092). DS were not different with or without antibiotic use with the probiotics. Only B. bifidum had an effect on self-reported stress scores (P=0.0086). The self-reported stress score was also dependent on hours of sleep per day where it decreased by 0.13 for each additional hour of sleep. During a stressful period, B. bifidum R0071 decreases DS and self-reported stress scores. This trial was registered at clinicaltrials.gov as NCT01709825.

OBJECTIVE:This study determined whether older adults who consumed a probiotic mixture would have a greater proportion of circulating CD4+ lymphocytes, altered cytokine production, and a shift in intestinal microbiota toward a healthier microbial community. METHODS:Participants (70 ± 1 years [mean ± SEM]; n = 32) consumed a probiotic (Lactobacillus gasseri KS-13, Bifidobacterium bifidum G9-1, and Bifidobacterium longum MM2) or a placebo twice daily for 3 weeks with a 5-week washout period between intervention periods. Blood and stools were collected before and after each intervention. The percentage of circulating CD4+ lymphocytes and ex vivo mitogen-stimulated cell cytokine production were measured. In stools, specific bacterial targets were quantified via quantitative polymerase chain reaction (qPCR) and community composition was determined via pyrosequencing. RESULTS:During the first period of the crossover the percentage of CD4+ cells decreased with the placebo (48% ± 3% to 31% ± 3%, p < 0.01) but did not change with the probiotic (44% ± 3% to 42% ± 3%) and log-transformed concentrations of interleukin-10 increased with the probiotic (1.7 ± 0.2 to 3.4 ± 0.2, p < 0.0001) but not the placebo (1.7 ± 0.2 to 2.1 ± 0.2). With the probiotic versus the placebo a higher percentage of participants had an increase in fecal bifidobacteria (48% versus 30%, p < 0.05) and lactic acid bacteria (55% versus 43%, p < 0.05) and a decrease in Escherichia coli (52% versus 27%, p < 0.05). Several bacterial groups matching Faeacalibactierium prausnitzii were more prevalent in stool samples with the probiotic versus placebo. CONCLUSIONS:The probiotic maintained CD4+ lymphocytes and produced a less inflammatory cytokine profile possibly due to the changes in the microbial communities, which more closely resembled those reported in healthy younger populations.

The aim of the studies was to determine the effects of calcium carbonate and calcium phosphate supplementation on faecal Lactobacillus spp., with and without a probiotic supplement, in healthy adults. Study 1 comprised of a randomised, double-blind, crossover design; participants (n=15) received 2 capsules/d of 250 mg elemental calcium as calcium carbonate (Ca1) and calcium phosphate (Ca2) each for 2-week periods, with 2-week baseline and washout periods. Study 2 was a randomised, double-blind, crossover design; participants (n=17) received 2 capsules/d of Lactobacillus helveticus R0052 and Lactobacillus rhamnosus R0011 (probiotic) alone, the probiotic with 2 capsules/d of Ca1, and probiotic with 2 capsules/d of Ca2 each for 2-week periods with 2-week baseline and washout periods. In both studies, stools were collected during the baseline, intervention and washout periods for Lactobacillus spp. quantification and qPCR analyses. Participants completed daily questionnaires of stool frequency and compliance. In Study 1, neither calcium supplement influenced viable counts of resident Lactobacillus spp., genome equivalents of lactic acid bacteria or stool frequency. In Study 2, faecal Lactobacillus spp. counts were significantly enhanced from baseline when the probiotic was administered with Ca2 (4.83±0.30, 5.79±0.31) ( P =0.02), but not with Ca1 (4.98±0.31) or with the probiotic alone (5.36±0.31, 5.55±0.29) (not significant). Detection of L. helveticus R0052 and L. rhamnosus R0011 was significantly increased with all treatments, but did not differ among treatments. There were no changes in weekly stool frequency. Calcium phosphate co-administration may increase gastrointestinal survival of orally-administered Lactobacillus spp.

Abstract Purpose: To determine which dietary factors contribute to microbiota variation and identify correlations with gut epithelial methylation pattern associated with the presence of high-risk polyps. Dietary exposures have long been suggested as important risk factors for cancers, especially those of the gastrointestinal tract. Diet is known to affect the commensal gut microbiota, which contributes important activities to the human host that are required for maintaining normal health. However, distortions in microbiota composition and associated microbial activities might contribute to disease processes including colorectal carcinogenesis. High-throughput sequencing based microbiota studies have revealed a large degree of intra- and inter-individual variation, the sources of which are currently not well understood. As part of this project we explored various ‘Big Data’ approaches that can facilitate an efficient mining of the large dataset generated by combining dietary intake data with output from multiple high-throughput technologies. We hypothesized that specific nutrients can affect microbiota composition, and that microbiota correlates with an epithelial methylation pattern associated with CRC risk. Methods: The study included 126 human healthy volunteers undergoing a screening colonoscopy. Dietary habits were assessed using 4-day food records, FFQ (Block 98) and Meat Module Questionnaire. Microbiota composition was examined in a stool samples collected before colonoscopy and in multiple biopsy samples retrieved during the screening procedure. The colonoscopy results were used for ascertainment of polyp status. Microbiota composition was analyzed in DNA extracted from stool and biopsy samples using a 16S rRNA based approach. In addition, in a subset of 39 stool samples a shotgun metagenomic analysis was undertaken. Sequence reads were binned into operational taxonomic units (OTUs) using ESPRIT and further analyzed using our in house analytical pipeline as well as publicly available platforms such as QIIME. We then performed a discriminant analysis to identify a microbiota pattern associated with polyp prevalence. Methylation analysis was performed in DNA extracted from biopsies obtained from six high-risk cases and six matched controls using the Infinium HumanMethylation450 beadchip. We used regression analysis approaches as well as the popular apriori rule-mining algorithm to identify associations in the complex heterogeneous dataset. Results: We observed microbiota associations with dietary intake as well as differences in the presence of OTUs in subjects with and without polyps (p<0.01), with the most significant of these differences detected in subjects with high-risk polyps. After adjustment for age, body mass index, gender, race and total caloric intake dietary fiber and vitamin D intake correlated with potentially beneficial butyrate producing bacteria. A predictive model based on 27 OTUs identified subjects with at least one polyp (AUC =0.81). Methylation status at multiple sites was associated with polyp status. Methylation at a site in HCA-DRB-1, a MHC gene, strongly correlated with the presence the microbial taxon Alistipes (p=0.0006). Our rule mining approach was able to identify associations between multiple layers of heterogenous data. Conclusions: Our observations suggest complex interactions between diet and microbiota that correlate with epithelial methylation pattern and CRC risk. An efficient mining of this and other large datasets, generated by combining multiple layers of complex data, will require adaptation of ‘Big Data’ analytical approaches that can handle the unique characteristics of high dimensional but sparse data. Citation Format: Tyler Culpepper, Maria Ukhanova, Lusine Yaghjyan, Samantha Sites, Yijun Sun, Prosperi Mattia, Volker Mai. Correlations between diet, gut microbiota and epithelial methylation pattern with CRC risk [abstract]. In: Proceedings of the Thirteenth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2014 Sep 27-Oct 1; New Orleans, LA. Philadelphia (PA): AACR; Can Prev Res 2015;8(10 Suppl): Abstract nr A31.

Aging is associated with dysregulated immune function. It was hypothesized that ingestion of a probiotic would induce a beneficial shift in gut microbiota composition resulting in a greater proportion of T helper (Th) cells, increased Th1 cytokine production, and decreased inflammation. In a 13‐wk study, older adults (70±1 y; n=32) consumed a probiotic mix or a placebo twice daily for 3 wks with a 5‐wk washout. Blood and stools were collected at baseline and post intervention. After the intervention, several bacterial groups matching Faeacalibactierium prausnitzii, a species inversely associated with inflammatory GI disease, were more prevalent in stool samples with the probiotic vs. placebo. Percentages of peripheral Th cells were higher after probiotic (41% ± 2%, mean ± SEM) vs. placebo (35% ± 3%). After in vitro mitogen stimulation of T cells, IFN‐γ (6.9 ± 0.2 vs. 5.5 ± 0.4, probiotic vs. placebo, log transformed means ± SEM, pg/mL), IL‐10 (4.2 ± 0.2 vs. 2.9 ± 0.3), and IL‐2 (4.7 ± 0.2 vs. 3.7 ± 0.4) concentrations were higher with ingestion of the probiotic. These data suggest that ingestion of this probiotic mix results in a potentially beneficial shift in gut microbiota composition at the individual species level and may impact cell‐mediated immunity in older adults.Grant Funding Source: Wakunaga of America Co. Ltd.

Resistant maltodextrin (RM) is a novel soluble, nonviscous dietary fiber. Its metabolizable energy (ME) and net energy (NE) values derived from nutrient balance studies are unknown, as is the effect of RM on fecal microbiota. A randomized, placebo-controlled, double-blind crossover study was conducted (n = 14 men) to determine the ME and NE of RM and its influence on fecal excretion of macronutrients and microbiota. Participants were assigned to a sequence consisting of 3 treatment periods [24 d each: 0 g/d RM + 50 g/d maltodextrin and 2 amounts of dietary RM (25 g/d RM + 25 g of maltodextrin/d and 50 g/d RM + 0 g/d maltodextrin)] and were provided all the foods they were to consume to maintain their body weight. After an adaptation period, excreta were collected during a 7-d period. After the collection period, 24-h energy expenditure was measured. Fluorescence in situ hybridization, quantitative polymerase chain reaction, and 454 titanium technology–based 16S rRNA sequencing were used to analyze fecal microbiota composition. Fecal amounts of energy (544, 662, 737 kJ/d), nitrogen (1.5, 1.8, 2.1 g/d), RM (0.3, 0.6, 1.2 g/d), and total carbohydrate (11.1, 14.2, 16.2 g/d) increased with increasing dose (0, 25, 50 g) of RM (P < 0.0001). Fat excretion did not differ among treatments. The ME value of RM was 8.2 and 10.4 kJ/g, and the NE value of RM was −8.2 and 2.0 kJ/g for the 25 and 50 g/d RM doses, respectively. Both doses of RM increased fecal wet weight (118, 148, 161 g/d; P < 0.0001) and fecal dry weight (26.5, 32.0, 35.8 g/d; P < 0.0001) compared with the maltodextrin placebo. Total counts of fecal bacteria increased by 12% for the 25 g/d RM dose (P = 0.17) and 18% for the 50 g/d RM dose (P = 0.019). RM intake was associated with statistically significant increases (P < 0.001) in various operational taxonomic units matching closest to ruminococcus, eubacterium, lachnospiraceae, bacteroides, holdemania, and faecalibacterium, implicating RM in their growth in the gut. Our findings provide empirical data important for food labeling regulations related to the energy value of RM and suggest that RM increases fecal bulk by enhancing the excretion of nitrogen and carbohydrate and the growth of specific microbial populations.

Background Recent advances in our understanding of the contributions of gut microbiota to host metabolism have renewed research interest in how distortions in microbiota activities correlate with health status. While extensive microbiota surveys have already revealed the complexity of microbiota dynamics there is a need to better understand mechanisms through which microbiota can affect host metabolism. We have recently shown that the amount of energy available to the host after intake of resistant maltodextrin varies among individuals and is associated with microbiota. Here we hypothesized that microbiota i) is associated with dietary habits, ii) correlates with colorectal cancer (CRC) risk and iii) can affect the methylation status in nearby gut epithelium, which might be a mechanism contributing to colorectal carcinogenesis.

A probiotic formulation of Enterococcus faecium R0026 and Bacillus subtilis R0179 has been evaluated in previous clinical trials. However, B. subtilis R0179 has not been evaluated as a single probiotic strain or in combination with other strains at doses higher than 0.1×109 cfu. To establish oral dose-response tolerance and gastrointestinal (GI) viability of B. subtilis R0179, a randomised, double-blind, placebo-controlled trial in healthy adults (n=81; 18-50 years old) was conducted. Participants received B. subtilis R0179 at 0.1, 1.0 or 10×109 cfu/capsule/day or placebo for four weeks. General wellness was assessed using a daily questionnaire evaluating GI, cephalic, ear-nose-throat, behavioural, emetic, and epidermal symptoms. GI symptoms were further evaluated using a weekly gastrointestinal symptom rating scale (GSRS). GI transit viability of B. subtilis R0179 was assessed by plating and microbiota analysis by 16S rRNA at baseline, week 4 of the intervention and washout. General wellness and GI function were not affected by oral consumption of B. subtilis R0179 at any dose. Daily questionnaire syndrome scores were not different from baseline and did not exceed a clinically significant score of 1. GSRS syndrome scores were not different from baseline and ranged from 1.1±0.1 to 1.9±0.2. Faecal viable counts of B. subtilis R0179 demonstrated a dose response: the placebo group (1.1±0.1 log10 cfu/g) differed from 0.1×109 (4.6±0.1 log10 cfu/g), 1×109 (5.6±0.1 log10 cfu/g) and 10×109 (6.4±0.1 log10 cfu/g) (P<0.0001). No significant changes in phyla were observed, but sequence reads binned to multiple operational taxonomic units matching closest to Ruminococci increased during probiotic supplementation. B. subtilis R0179 survives passage through the human GI tract and is well tolerated by healthy adults at intakes from 0.1 to 10×109 cfu/day. The trial has been registered at www.clinicaltrials.gov under NCT01802151.