Prebiotics and the health benefits of fiber: current regulatory status, future research, and goals.

First defined in the mid-1990s, prebiotics, which alter the composition and activity of gastrointestinal (GI) microbiota to improve health and well-being, have generated scientific and consumer interest and regulatory debate. The Life Sciences Research Organization, Inc. (LSRO) held a workshop, Prebiotics and the Health Benefits of Fiber: Future Research and Goals, in February 2011 to assess the current state of the science and the international regulatory environment for prebiotics, identify research gaps, and create a strategy for future research. A developing body of evidence supports a role for prebiotics in reducing the risk and severity of GI infection and inflammation, including diarrhea, inflammatory bowel disease, and ulcerative colitis as well as bowel function disorders, including irritable bowel syndrome. Prebiotics also increase the bioavailability and uptake of minerals and data suggest that they reduce the risk of obesity by promoting satiety and weight loss. Additional research is needed to define the relationship between the consumption of different prebiotics and improvement of human health. New information derived from the characterization of the composition and function of different prebiotics as well as the interactions among and between gut microbiota and the human host would improve our understanding of the effects of prebiotics on health and disease and could assist in surmounting regulatory issues related to prebiotic use.

A defined intestinal colonization microbiota for gnotobiotic pigs

Maximising the ability of piglets to survive exposure to pathogens is essential to reduce early piglet mortality, an important factor in efficient commercial pig production. Mortality rates can be influenced by many factors, including early colonization by microbial commensals. Here we describe the development of an intestinal microbiota, the Bristol microbiota, for use in gnotobiotic pigs and its influence on synthesis of systemic immunoglobulins. Such a microbiota will be of value in studies of the consequences of early microbial colonization on development of the intestinal immune system and subsequent susceptibility to disease. Gnotobiotic pig studies lack a well-established intestinal microbiota. The use of the Altered Schaedler Flora (ASF), a murine intestinal microbiota, to colonize the intestines of Caesarean-derived, gnotobiotic pigs prior to gut closure, resulted in unreliable colonization with most (but not all) strains of the ASF. Subsequently, a novel, simpler porcine microbiota was developed. The novel microbiota reliably colonized the length of the intestinal tract when administered to gnotobiotic piglets. No health problems were observed, and the novel microbiota induced a systemic increase in serum immunoglobulins, in particular IgA and IgM. The Bristol microbiota will be of value for highly controlled, reproducible experiments of the consequences of early microbial colonization on susceptibility to disease in neonatal piglets, and as a biomedical model for the impact of microbial colonization on development of the intestinal mucosa and immune system in neonates.

Expression of cyclooxygenase-2 in intestine of pigs of different ages and hygiene status

Prostaglandins (PG) are bioactive lipids derived from the metabolism of membrane polyunsaturated fatty acids (PUFA), and play important roles in a number of biological processes including cell division, immune responses and wound healing. Cyclooxygenase (COX) is the key enzyme in PG synthesis from arachidonic acid. The hypothesis of the present study was that expression of COX-2 in porcine intestine was dependent on the microbial load and the age of piglets. Piglets were obtained from sows raised either on outdoor free-range farms or on indoor commercial farms, and littermates were divided into three treatments: One group of piglets suckled the sow, a second group was put into an isolator and fed a milk formula, and a third group was put into the isolator fed milk formula and injected with broad spectrum antibiotics. Samples were collected from the 75% level of the small intestine at day 5, 28 and 56 of age. Tissue section from four piglets from each of these six treatment groups was analysed by immunofluorescence for COX-2 and type-IV collagen (basement membrane, defining lamina propria (LP)). Image analysis was used to determine the number of positive pixels expressing LP and epithelial COX-2. COX-2 expressing cells were observed in LP and epithelium in all porcine intestinal samples. When analysing images obtained on day 28, injection of antibiotics seemed to reduce the COX-2 expression in intestinal samples of piglets when compared to other treatments (P=0.053). No significant effect of farm, treatments or age of piglets was observed on COX-2 expressing data when analysing all data of images obtained at day 28 and 56. By double-labelling experiments, COX-2 was found not to be expressed on cell co-expressing CD45, CD16, CD163 or CD2, thus indicating that mucosal leukocytes, including dendritic cells, macrophages and NK cells did not express COX-2. Future research should investigate the role of COX-2 expression in the digestive tract in relation to pig health.

Environmentally-acquired bacteria influence microbial diversity and natural innate immune responses at gut surfaces

Background: Early microbial colonization of the gut reduces the incidence of infectious, inflammatory and autoimmune diseases. Recent population studies reveal that childhood hygiene is a significant risk factor for development of inflammatory bowel disease, thereby reinforcing the hygiene hypothesis and the potential importance of microbial colonization during early life. The extent to which early-life environment impacts on microbial diversity of the adult gut and subsequent immune processes has not been comprehensively investigated thus far. We addressed this important question using the pig as a model to evaluate the impact of early-life environment on microbe/host gut interactions during development. Results: Genetically-related piglets were housed in either indoor or outdoor environments or in experimental isolators. Analysis of over 3,000 16S rRNA sequences revealed major differences in mucosa-adherent microbial diversity in the ileum of adult pigs attributable to differences in earlylife environment. Pigs housed in a natural outdoor environment showed a dominance of Firmicutes, in particular Lactobacillus, whereas animals housed in a hygienic indoor environment had reduced Lactobacillus and higher numbers of potentially pathogenic phylotypes. Our analysis revealed a strong negative correlation between the abundance of Firmicutes and pathogenic bacterial populations in the gut. These differences were exaggerated in animals housed in experimental isolators. Affymetrix microarray technology and Real-time Polymerase Chain Reaction revealed significant gut-specific gene responses also related to early-life environment. Significantly, indoorhoused pigs displayed increased expression of Type 1 interferon genes, Major Histocompatibility Complex class I and several chemokines. Gene Ontology and pathway analysis further confirmed these results.

Computational analysis of the LRRK2 interactome

LRRK2 was identified in 2004 as the causative protein product of the Parkinson’s disease locus designated PARK8. In the decade since then, genetic studies have revealed at least 6 dominant mutations in LRRK2 linked to Parkinson’s disease, alongside one associated with cancer. It is now well established that coding changes in LRRK2 are one of the most common causes of Parkinson’s. Genome-wide association studies (GWAs) have, more recently, reported single nucleotide polymorphisms (SNPs) around the LRRK2 locus to be associated with risk of developing sporadic Parkinson’s disease and inflammatory bowel disorder. The functional research that has followed these genetic breakthroughs has generated an extensive literature regarding LRRK2 pathophysiology; however, there is still no consensus as to the biological function of LRRK2. To provide insight into the aspects of cell biology that are consistently related to LRRK2 activity, we analysed the plethora of candidate LRRK2 interactors available through the BioGRID and IntAct data repositories. We then performed GO terms enrichment for the LRRK2 interactome. We found that, in two different enrichment portals, the LRRK2 interactome was associated with terms referring to transport, cellular organization, vesicles and the cytoskeleton. We also verified that 21 of the LRRK2 interactors are genetically linked to risk for Parkin- son’s disease or inflammatory bowel disorder. The implications of these findings are discussed, with particular regard to potential novel areas of investigation.

Measurement of the ratio of B+ and B-0 meson lifetimes

The ratio of the B+ and B-0 meson lifetimes was measured using data collected in 2002-2004 by the D0 experiment in Run II of the Fermilab Tevatron Collider. These mesons were reconstructed in B→μ(+)ν D*-X decays, which are dominated by B-0 and B→μ(+)ν(D) over bar X-0 decays, which are dominated by B+. The ratio of lifetimes is measured to be τ(+)/τ(0)=1.080± 0.016(stat)± 0.014(syst).

Search for the flavour-changing neutral current B-s(0)->mu+mu- in p(p)over-bar collisions at root s=1.96 TeV with the D0 dectector

We present the results of a search for the flavor-changing neutral current decay B-s(0)-->mu(+)mu(-) using a data set with integrated luminosity of 240 pb(-1) of p (p) over bar collisions at roots=1.96 TeV collected with the D0 detector in run II of the Fermilab Tevatron collider. We find the upper limit on the branching fraction to be B(B-s(0)-->mu(+)mu(-))less than or equal to5.0x10(-7) at the 95% C.L. assuming no contributions from the decay B-d(0)-->mu(+)mu(-) in the signal region. This limit is the most stringent upper bound on the branching fraction B-s(0)-->mu(+)mu(-) to date.

Search for first-generation scalar leptoquarks in p(p)over-bar collisions at root s=1.96 TeV

We report on a search for pair production of first-generation scalar leptoquarks (LQ) in p (p) over bar collisions at root s=1.96 TeV using an integrated luminosity of 252 pb(-1) collected at the Fermilab Tevatron collider by the D0 detector. We observe no evidence for LQ production in the topologies arising from LQ(LQ) over bar -> eqeq and LQ(LQ) over bar -> eq nu q, and derive 95% C.L. lower limits on the LQ mass as a function of beta, where beta is the branching fraction for LQ -> eq. The limits are 241 and 218 GeV/c(2) for beta=1 and 0.5, respectively. These results are combined with those obtained by D0 at root s=1.8 TeV, which increases these LQ mass limits to 256 and 234 GeV/c(2).

Measurement of the B-s(0) lifetime in the exclusive decay channel B-s(0)-> J/psi phi

Using the exclusive decay B-s(0)-->J/psi(mu(+)mu(-))phi(K+K-), we report the most precise single measurement of the B-s(0) lifetime. The data sample corresponds to an integrated luminosity of approximately 220 pb(-1) collected with the D0 detector at the Fermilab Tevatron Collider in 2002-2004. We reconstruct 337 signal candidates, from which we extract the B-s(0) lifetime, tau(B-s(0))=1.444(-0.090)(+0.098)(stat)+/-0.020(sys) ps. We also report a measurement for the lifetime of the B-0 meson using the exclusive decay B-0-->J/psi(mu(+)mu(-))K-*0(892)(K(+)pi(-)). We reconstruct 1370 signal candidates, obtaining tau(B-0)=1.473(-0.050)(+0.052)(stat)+/-0.023(sys) ps, and the ratio of lifetimes, tau(B-s(0))/tau(B-0)=0.980(-0.071)(+0.076)(stat)+/-0.003(sys).

Measurement of the WW production cross section in p(p)over-bar collisions at root(s)over-bar=1.96 TeV

We present a measurement of the W boson pair-production cross section in p(p) over bar collisions at a center-of-mass energy of root s=1.96 TeV. The data, collected with the Run II D0 detector at Fermilab, correspond to an integrated luminosity of 224-252 pb(-1) depending on the final state (ee, e mu, or mu mu). We observe 25 candidates with a background expectation of 8.1 +/- 0.6(stat)+/- 0.6(syst)+/- 0.5(lum) events. The probability for an upward fluctuation of the background to produce the observed signal is 2.3x10(-7), equivalent to 5.2 standard deviations. The measurement yields a cross section of 13.8(-3.8)(+4.3)(stat)(-0.9)(+1.2)(syst)+/- 0.9(lum) pb, in agreement with predictions from the standard model.

Search for anomalous heavy-flavor quark production in association with W bosons

We search for anomalous production of heavy-flavor quark jets in association with W bosons at the Fermilab Tevatron p(p) over bar Collider in final states in which the heavy-flavor quark content is enhanced by requiring at least one tagged jet in an event. Jets are tagged using one algorithm based on semileptonic decays of b/c hadrons, and another on their lifetimes. We compare e+jets (164 pb(-1)) and mu+jets (145 pb(-1)) channels collected with the D0 detector at root s = 1.96 TeV to expectations from the standard model and set upper limits on anomalous production of such events.