Investigation of the Ability of a Supplement to Increase Good Bacteria in the Human Intestine and Blood Sugar Levels

Prebiotics are highly effective and important for many applications in medicine. They are
not absorbed and do not contribute to human nourishment, but rather exert a profound effect
on the human bowel flora. The principal effect on the human bowel flora is to stimulate the
growth of Bifidobacterium and Lactobacillus species. These are benign organisms in that
they are seldom involved in infections or other pathological processes. However, numerous
health promoting benefits of these bacterial genera have been noted, with effects on
infectious and non-infectious disease. They induce host genes involved in innate immunity,
sometimes in collaboration with other elements of the gut flora such as Bacteroides
thetaiotaomicron and Lactobacillus casei. Animal studies 2 indicate a protective effect
against bacterial translocation from the GI tract. An extensive cataloguing of genomes of
bifidobacteria and other intestinal bacteria reveals that the bifids play a very significant
role in many important areas such as carbohydrate, amino acid, nucleotide, lipid, inorganic
ion transport and metabolism; energy production and conversion; cell wall and membrane
biogenesis; signal transduction mechanisms; transcription and translation; and defense
mechanisms .

The metabolic syndrome which includes non-alcoholic fatty liver disease, obesity, and
insulin resistance has responded to prebiotics in rodent trials; it is estimated that 20-30%
of populations in developed countries have this disease. Parnell and Reimer have
demonstrated that prebiotics oligofructose supplementation has the potential to promote
weight loss and improve glucose regulation in overweight adults likely through suppressed
ghrelin and enhanced PYY1.

The investigators have completed a study at UCLA studying the effects of XOS on human
microbiota. There were three phases including a 2-week run-in period, 8-week intervention
period, and a 2-week washout. Thirty two volunteers were randomly assigned to take a
supplement containing 1.4 g XOS (1.4g XOS 70P), 2.8 g XOS (2.8g XOS 70P) or placebo.

Total of 32 subjects were enrolled into the study. One subject from the placebo group and
one from the low dose group dropped out of the study for non specific gastrointestinal
complaint.

Altogether, 120 stool samples were received from 11 low dose subjects, 9 high dose subjects,
and 10 placebo group subjects. Bacterial culture was performed from all the 120 stool
samples received. One placebo group subject, two high dose group subjects, and two low dose
group subjects were excluded from the analyses because of compromised specimen quality.

The Bifidobacterium counts of the subjects after high dose XOS intervention was
significantly higher from the base line at 4, 8, and 10 weeks. Similarly, the increase of
Bifidobacterium counts was significantly higher in the high XOS group at 4 weeks compared to
the low XOS group. The low XOS group had significantly higher Bifidobacterium counts
compared to the placebo group subjects at 8 and 10 weeks. The total anaerobic flora counts
and to some degree the total aerobic flora counts of the subjects after high dose XOS
intervention were significantly higher from the base-line at 4 and 8 weeks. The mean changes
of total anaerobic flora counts were significantly higher in the high XOS group at 4, 8, and
10 weeks compared to the placebo group. Interestingly, also Bacteroides fragilis group
counts of the subjects after high dose XOS intervention were significantly higher from the
base line at 4, 8, and 10 weeks and the mean changes were significantly higher in the high
XOS group compared to the low and placebo groups. There were no major significant
differences in the counts of Lactobacillus sp., Enterobacteriaceae, and clostridia between
the three dose groups evaluated

The XOS was tolerated well by all study subjects (normal adult). At base-line, most of the
subjects had relatively high counts of Bifidobacterium species with lower counts of
Lactobacillus species. Several different species of each of these genera were present.
Bifidobacterium species showed significant increases in counts in subjects on XOS
supplementation, with the higher dose group showing significantly greater increases than the
lower dose group. Both high and low dose groups showed significantly higher counts of
Bifidobacterium sub-species than the placebo group. Lactobacillus species counts did not
increase significantly in subjects on XOS supplementation. There was a statistically
significant increase in the counts of the B. fragilis group at all time periods for the high
dose XOS group. As mentioned before, bifidobacteria may induce host genes involved in innate
immunity in collaboration with other elements of the gut flora such as B. fragilis group.
The increase in the counts of the "total anaerobes" reflects the increased counts in the
Bifidobacterium and B. fragilis group, both of which are anaerobic. Some Bifidobacterium
species may grow aerobically as well, which may be reflected in the modest increase of the
aerobic counts in the high dose XOS group. In this study, the XOS supplementation had no
significant effect on stool pH or SCFAs.

The investigators propose to conduct a 10 week, randomized, placebo controlled, two arm
study with 20 subjects with metabolic syndrome. Subjects will be randomly assigned to take
a powder mixture containing 2.8 g XOS (2.94g XOS 95P) or placebo.

Inclusion Criteria:

1. Age 20-70 years of age at screen

2. BMI between 27 to 35

3. Fasting glucose level >100 mg/dL or >200 mg/dL at 1 hour after ingesting of 75 grams
of glucose

4. Subjects must read and sign the Institutional Review Board-approved written informed
consent prior to the initiation of any study specific procedures or enrollment. A
subject will be excluded for any condition that might compromise the ability to give
truly informed consent

Exclusion Criteria:

1. Any subject with a history of diabetes mellitus on medications, or other serious
medical condition, such as chronic hepatic or renal disease, bleeding disorder,
congestive heart disease, chronic diarrhea disorders, myocardial infarction, coronary
artery bypass graft, angioplasty within 6 months prior to screening, current
diagnosis of uncontrolled hypertension (defined as systolic BP>160mmHg, diastolic
BP>95mmHg), active or chronic gastrointestinal disorders, bulimia, anorexia, or
endocrine diseases (except thyroid disease requiring medication) as indicated by
medical history or routine physical examination.

2. Any subject with a screening laboratory value outside of the laboratory normal range
that is considered clinically significant for study participation by the
investigator.

3. Any subject who currently uses tobacco products.

4. Any history of gastrointestinal disease except for appendectomy

5. No antibiotics or laxatives use during the 2 months before the study.

6. Any subject who is unable or unwilling to comply with the study protocol.