Note: Descriptions are shown in the official language in which they were submitted.
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TITLE: KONJAC MANNAN AND POLYSACCHARIDE
COMPOSITIONS AND USES THEREOF
FIELD OF THE INVENTION
The invention relates to compositions comprising Konjac-Mannan,
Ginseng or both and methods of use and uses of these compositions, such as
in lowering blood glucose including post-prandial and long-term effects. In
one
aspect this invention is in the field of glucose and other heart disease risk
factors management. In one aspect the invention is concerned with dietary
approaches to such management. In another aspect it is concerned with
compositions and methods of reducing blood glucose, specifically
BACKGROUND OF THE INVENTION
Abnormal glucose tolerance and insulin resistance are related to
multiple cardiovascular risk factors especially reduced HDL, elevated serum
triglycerides and hypertension (Liese et al. (1998)). When clustered these
abnormalities increase the risk of coronary heart disease (CHD) morbidity and
mortality, an effect that is independent of other conventional risk factors
(Trevisan et al. (1998)). Co-ocurrence is usually present in insulin-
insensitive
individuals (Himswarth (1936)) and is often described in relation to visceral
adiposity (Haffner et al. (1986)) and lack of physical activity (Helmrich
1991)).
The estimated prevalence ranges from 3% (Trevisan et al. (1998)) to
approximately 30% (Liese et al. (1998); Reaven (1994)) depending on how
this insulin resistance-dislipidemic syndrome is defined and in which
population it is measured.
Hyperglycemia and diabetes are strong and independent risk factors of
both all-cause and cardiovascular (CVD) mortality (Wing et al. (1998)). These
links are more pronounced when the diabetes is associated with other
unfavorable risk factors such as hyperlipidemia (Goldsmith et al. (1994)),
hypertension (Burt et al. (1995)), or a cluster of metabolic disorders
(Stamler
et al. (1993)). Since people with diabetes have almost twice the risk of dying
from CVD (69.6%) compared to people in the general U.S. population (Gu et
al. (1998)), the control of high glucose levels and other concomitant coronary
heart disease (CHD) risk factors represents the most effective approach to
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prevention (Savage (1996)). The importance of stronger nutrition-hygienic
measures has been stressed repeatedly for the public at large (Stamler et al.
(1993); National Cholesterol Education Program: Second report of the expert
panel on detection, evaluation, and treatment of high blood cholesterol in
adults (adult treatment panel II). Circulation. 1994; 89:1333-1445)). When
these measures prove inadequate, an aggressive drug therapy is often
required to meet the conventional treatment guidelines (National Cholesterol
Education Program: Second report of the expert panel on detection,
evaluation, and treatment of high blood cholesterol in adults (adult treatment
panel II). Circulation. 1994; 89:1333-1445)). In the general population, this
approach has been shown to be effective in lowering both the prevalence of
hypertension (Burt et al. (1995)) and serum cholesterol levels (Johnson et al.
(1993)), but has not reduced the incidence of diabetes (Harris et al. (1998)).
Although it has been extensively described (Liese et al. (1998);
Trevisan et al. (1998; Himswarth (1936); Haffner et al. (1986); Helmrich et
al.
(1994)), followed-up (Reaven (1994)), and had its prevalence determined
(1,2), no specific recommendations for treatment of this syndrome have been
proposed by health agencies. In practice, initial therapy of individual risk
factors such as moderate dyslipidemia, hypertension or hyperglycemia is
nonpharmacological. Treatment will often include behavioral changes to
reduce body weight, increase physical activity, and moderate alcohol
consumption. To achieve nutritional goals, there are three main approaches:
a high-carbohydrate/low-fat diet (National Cholesterol Education Program:
Second report of the expert panel on detection, evaluation, and treatment of
high blood cholesterol in adults (adult treatment panel II) Circulation
89:1333-
1445 (1994)), sharing calories between monounsaturated fat and complex
carbohydrate at the expense of saturated fat (American Diabetes Association
(ADA): Nutrition Recommendations and principles for people with diabetes
mellitus. Diabetes Care 22:s42-s43 (1999)), or supplementing a high-
carbohydrate/low-fat diet with exercise (Stefanick et al. (1998)).
Tighter fasting and postprandial glycemic control results in a
considerable reduction in CHD and all-cause mortality (Wei et al. (1998)), as
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well as fewer long-term microvascular complications both in type 1 (DCCT
Research Group: The effect of intensive treatment of diabetes on the
development and progression of long-term complications in insulin-dependent
diabetes mellitus. The diabetes control and complications trial. New Engl J
Med 329:977-986 (1993)) and type 2 diabetes (UK Prospective Diabetes
Study (UKPDS) Group: Effect of intensive blood-glucose control with
metformin on complications in overweight patients with type 2 diabetes:
UKPDS 34. Lancet 352:854-865, 1998). Effective dietary strategies shown to
decrease (This OUT postprandialThis OUT) plasma glucose excursions
include the use of high fibre and low glycemic index diets (Wolever et al.
(1992); Jenkins et al. (1994)). The mechanism is presumed to involve slowing
carbohydrate absorption (Jenkins et al. (1994)). Based on recent population
studies these types of diets have been shown to have a protective role in
preventing diabetes (Salmeron et al., Diabetes Care 20:545-550 (1997);
Salmeron et al., JAMA 277:472-477 (1997)) and CHD (18). In the case of
clinical studies however, it is the viscous water-soluble fibers, which
increase
the viscosity of digesta in the human gut (Eastwood et al. (1992)) that reduce
glucose and lipid CHD risk factors (Anderson et al. (1986)). Whether soluble
fibre is able to reduce a cluster of risk factors is speculative. Studies
using
soluble fibre as an adjunct to conventional treatment in individuals with two
or
more major CHD risk factors are scarce (Uuistupa et al. (1984)).
Evidence suggests that fibre may also be used in a therapeutic role.
Recent epidemiological findings confirm the relationship between high dietary
fiber intake and lower risk of developing both diabetes (Salmeron et al.
(1997); Salmeron et al. (1997)) and CHD (Rimm et al. (1996)). Soluble
dietary fiber, in particular, has been shown clinically to reduce the need for
insulin, (Landin et al. (1992)) improve glycemia (Aro et al. (1981)), and
reduce
serum LDL-cholesterol (Brown et al. (1999)). Its viscosity is proposed as an
important mechanistic factor (Jenkins et al. (1978)). However, to date, there
is no clearly effective composition or method for reducing blood glucose.
SUMMARY OF THE INVENTION
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The present inventor has determined that the addition of high-viscosity
fiber, in the form of a konjac-mannan mixture, or ginseng, or a composition
comprising a konjac-mannan mixture and ginseng to a diet of an animal
enhances conventional treatment outcomes, such as for diabetes and
coronary heart disease, assessed primarily by total/HDL cholesterol,
fructosamine, and sBP and secondarily by total, LDL, and HDL cholesterol;
apolipoprotein A-1 (Apo A-1), B (Apo B) and their ratio; glucose; insulin; and
systolic blood pressure (dBP).
Accordingly, in one aspect the present invention provides a
composition of matter for reducing blood glucose comprising a konjac-
mannan mixture. In another aspect of the invention, a sufficient amount of the
konjac-mannan mixture, when given to an animal preferably at an appropriate
time, reduces blood glucose in the animal, preferably postprandial blood
glucose. Preferably the konjac-mannan mixture comprises konjac-mannan
and a substance capable of increasing the viscosity of konjac-mannan from
50% to about 250% of konjac-mannan alone. More preferably the substance
comprises about 5% to about 45% by weight of one or more such substances,
preferably polysaccharides.
According to one embodiment a composition as just mentioned
comprises as the one or more polysaccharides xanthan, carragenan, acetan,
guar, or xyloglucana.
According to another embodiment a composition according to the
invention comprises consitituents with the particle size larger than about
1,000
angstroms. In one aspect such a composition is used in controlling cholesterol
levels, preferably lowering cholesterol levels.
Preferably, compositions according to the invention are formulated into
a liquid, powder or formulated as part of a food. However, in another
embodiment the compositions of the invention are formulated into pills,
capsules and tablets
According to another aspect the present invention provides a method
for reducing blood glucose in an animal comprising administering to the
animal a sufficient amount of a konjac-mannan mixture at an appropriate time
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in order to reduce bood glucose, preferably postprandial blood glucose, in the
animal. Preferably the konjac-mannan mixture comprises konjac-mannan and
a substance capable of increasing the viscosity of konjac-mannan to from
about 50% to about 250% of konjac-mannan alone, preferably the substance
comprises from about 5% to about 45% by weight of one or more
polysaccharides, more preferably the one or more polysaccharides are
selected from the group consisting of xanthan, carragenan, acetan, guar, or
xyloglucana.
According to one embodiment of the method, the particle size of the
constituents of a mixture of the invention are larger than about 1000
angstroms.
According to another embodiment of the invention, a mixture according
to the invention is administered orally in an amount of about 1 to about 4
grams per day, preferably the mixture is administered either prior to a meal
or
during the meal.
According to yet another embodiment of the method of the invention
the administration of a mixture according to the invention is by a liquid, a
powder, or as a part of a food product. In another embodiment administration
is by way of tablet, capsule or pill.
According to another aspect of the invention, there is provided a
composition of matter for reducing blood glucose comprising ginseng in one
aspect of the invention a sufficient amount of ginseng, when given to an
animal at an appropriate time reduces blood glucose, preferably postprandial
blood glucose, in the animal.
According to another aspect of the invention, there is provided
composition of matter for reducing blood glucose comprising an extract of
ginseng, preferably American ginseng, a sufficient amount of which when
given to an animal at an appropriate time reduces blood glucose in the
animal. According to any of the ginseng compositions,whether root or extract,
the ginseng is comprised of a ratio of protopanaxadiols (diols) relative to
protopanaxatriols (triols) that is preferably greater than about 1.0, most
preferably about 1.5 or greater and more preferably about 1.90 or greater. In
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another embodiment, the diol/triol ratio is about 2.4 or greater. In one
embodiment, the composition is formulated into a liquid, powder or formulated
as part of a food. In another embodiment the composition is formulated into
tablets, capsules or pills.
According to another aspect of the present invention there is provided
a method for reducing blood glucose in an animal comprising administering to
the animal a sufficient amount of ginseng at an appropriate time in order to
reduce blood glucose in the animal. In another aspect, there is provided a
method for reducing blood glucose in an animal comprising administering to
the animal a sufficient amount of an extract of ginseng at an appropriate time
in order to reduce blood glucose in the animal. According to either of these
latter mentioned methods, preferably the ginseng or extract is administered
before a meal or with a meal, more preferably the administration before meal
occurs from about I to about 180 minutes before the meal.
is According to one embodiment of the method of the invention the
amount of ginseng or extract of ginseng is at least about 1000 mg per
administration. In another embodiment, the amount of ginseng administered is
between 1-9 g, preferably 1-4g and most preferably 1-3 g.
According to another embodiment of the method of the invention the
ginseng comprises a ratio of diols/triols of greater than about 1.0, most
preferably about 1.5 or greater and more preferably about 1.90 or greater. In
another embodiment, the triols/diols ratio is about 2.4 or greater.
According to yet another embodiment of the method of the invention
the composition is administered as a food, a powder, or a liquid. In another
embodiment the composition is formulated into tablets, capsules or pills.
According to another aspect of the present invention there is provided
a composition of matter for reducing blood glucose comprising a konjac-
mannan mixture of the invention and ginseng. In one embodiment of the
invention a sufficient amount of konjac mannan and ginseng which when
given to an animal at an appropriate time reduces blood glucose in the
animal. Preferably the konjac-mannan mixture comprises konjac-mannan and
a substance capable of increasing the viscosity of konjac-mannan from 50%
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to about 250% of konjac-mannan alone. Preferably the substance comprises
about 10% to about 40% or one or more polysaccharides. More preferably the
one or more polysaccharides are xanthan, carragenan, acetan, guar, or
xyloglucana.
According to one embodiment according to this aspect of the invention
the particle size of constituents within the mixture is larger than about
1,000
angstroms.
According to another embodiment according to this aspect of the
invention the American ginseng is comprised of a ratio of diols/triols of
greater
than about 1.0, most preferably about 1.5 or greater and more preferably
about 1.90 or greater. In another embodiment, the diol/triol ratio is about
2.4
or greater. According to yet another embodiment according to this aspect of
the invention the composition is formulated into a liquid, powder or
formulated
as part of a food.
According to another embodiment, the ginseng used in the
compositions and methods fo the invention can be any ginseng or extract
thereof, for instance American, Asian, Chinese or other ginseng known in the
art. In another embodiment, the ginseng is preferably American ginseng. In
yet another embodiment of the invention , the ginseng or extract thereof has a
diol/triol ratio of about 1.0, most preferably about 1.5 or greater and more
preferably about 1.90 or greater. In another embodiment, the diol/triol ratio
is
about 2.4 or greater.
In another embodiment, the invention provides a method for
determining whether a ginseng or extract thereof would be useful in
controlling, modulating or lowering blood glucose, preferably postprandial
blood glucose, by determining the diol/triol ratio of the ginseng or extract
and
selected for one that has a diol/triol ratio of about 1.5 or greater. The
selected
extract can then be optionally tested for such activity using the traditional
tests
and assays described herein or known in the art. In another embodiment, the
selected ginseng can also be tested and selected for insulin, lipid profile,
cholesterol and blood pressure control, modulation or decrease. In yet
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another embodiment the selected ginseng can be tested and selected for
effectiveness in the treatment of diabetes and/or cardiovascular disease.
According to another aspect of the present invention the compositions
and methods of the invention can be applied to the treatment of long term
diabetes, heart disease, and syndrome X. In addition the compositions and
methods of the invention provide methods for increasing insulin sensitivity in
an animal and of treating type 2 diabetes as well as for reducing systolic
blood
pressure or blood cholesterol and other lipids and apolipoproteins
The details of the preferred embodiment of the present invention are
set forth in the accompanying drawings and the description below. Once the
details of the invention are known, numerous additional innovations and
changes will become obvious to one skilled in the art.
BRIEF DESCRIPTION OF THE DRAWINGS
The invention will be better understood with reference to the drawings
in which:
Figure 1 is a histogram illustrating the effect of various fibres including
Konjac mannan alone on glucose response in individuals in eleven individuals
in comparison with a Konjac-mannan mixture of the present invention.
Figure 2 is a graph illustrating the post-meal blood glucose response of
individuals histogram illustrating the effect of various fibres including
Konjac
mannan alone in comparison with a Konjac-mannan mixture of the present
invention.
Figure 3 is a bar graph illustrating the composition of the placebo
versus konjac biscuits used in Example 10.
Figure 4 illustrates the effect of konjac mannan versus placebo on
blood glucose levels as described in Example 10. Figure 4A is a linear graph
illustrating glucose response to a test breakfast before 3-week treatment and
Figure 4B is a linear graph illustrating glucose response to a test breakfast
after the 3 week treatment. Figure 4C is a bar graph illustrating glucose area
under the curve (AUC) before and after the 3-week treatment.
Figure 5 illustrates the effect of konjac mannan versus placebo on
blood insulin levels as described in Example 10. Figure 5A is a linear graph
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illustrating insulin response to a test breakfast before 3-week treatment and
Figure 5B is a linear graph illustrating insulin response to a test breakfast
after
the 3 week treatment. Figure 5C is a bar graph illustrating insulin area under
the curve (AUC) before and after the 3-week treatment.
Figure 6 illustrates the effect of highly refined konjac mannan versus
the konjac mixture of the invention as described in Example 11. Figure 6A is a
linear graph illustrating absolute blood glucose levels. Figure 6B is a linear
graph illustrating incremental blood glucose levels. Figure 6C is a bar graph
illustration glucose area under the curve.
io Figure 7 is a linear graph illustrating the results of Example 12 for the
effect of konjac mannan, American ginseng and both konjac mannan and
American ginseng on systolic blood pressure.
Figure 8 illustrated the effect of konjac mannan and ginseng treatment
versus the separate treatments on blood glucose. Figure 8 A and B are linear
graphs illustrating blood glucose levels versus time and changes therein as
described in Example 13.
Figure 9 is a linear graph illustrating insulin levels versus time using
various treatment regimes: placebo, ginseng, konjac mannan mix, and konjac
mannan mix and ginseng. Figure 9A shows insulin day profile. Figures 9B
shows incremental insulin day profile.
Figure 10 is a bar graph illustrating the effect of various ginseng
preparations with various diol/triol ratios on postpranial blood glucose
levels.
Figure 11 illustrates a comparison of incremental change (linear graphs
11A, C, and E) and area under the curve (AUC) (Figures 118, D, and F) in
plasma glucose (Figures 11A and B), insulin (Figures 11 C and D) and nitric
oxide (Nox) (Figures 11 E and F).
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS OF THE
INVENTION
As described above, the present invention is related to
compositions and methods for reducing blood glucose. In particular, the
present inventor has found that a konjac mannan mixture, a ginseng
composition, or a composition comprising konjac mannan and ginseng are
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effective in the reduction of blood glucose. In another embodiment, the
compositions of the invention are effective in controlling or modulating the
lipid
profile of an animal, blood pressure and insulin levels. The compositions fo
the invention A enhances conventional treatment outcomes, such as for
diabetes and coronary heart disease, assessed primarily by total/HDL
cholesterol, fructosamine, and sBP and secondarily by total, LDL, and HDL
cholesterol; apolipoprotein A-1 (Apo A-1), B (Apo B) and their ratio; glucose;
insulin; and systolic blood pressure (dBP).
As used herein "about" when referring to a value includes amounts that
within the scope of scientific certainty or uncertainty. Such amounts would
include amounts that are effectively equivalent to the values stated. "About"
when used in relation to diol/triol ratios means plus or minus 10% of the
value.
As used herein "animal" means any member of the animal kingdom
including preferably humans.
As used herein "postprandial" means after any food intake.
As used herein "sufficient amount" or "effective amount" means an
amount of a composition, substance or reactant to give an observable result,
including desired results.
As used herein "appropriate time" means at a time at which
administration of a substance or composition provides an observable result.
As used herein "prior to a meal" means at any time after a meal and
before a subsequent meal.
As used herein "during a meal" means at any time after the
commencement of consumption of one or more pieces of food by an animal,
and can be coincident with commencement, and before the end of
consumption of all food consumed by the animal, at one sitting or occasion
and can be coincident with completion of consumption or immediately
thereafter.
As used herein "a food" means any substance or composition of
substances or compounds which are consumed by an animal, preferably for
some nutritional value.
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As used herein "a meal" means the consumption of one or more
morsels or pieces of a food in a sitting where a sitting is the time taken to
consume the one or more morsels or pieces of a food.
As used herein, "diol/triol ratio(s)" or "diols/triols" refers to
protopanaxadiols (diols) relative to protopanaxatriols (triols).
Konjac Mannan
Konjac (Amorphophalus Konjac C. Koch) is a perennial plant belonging
to the family Araceae. "Konnyaku", which is made from the tuber of this plant,
has been used traditionally for food in Japan for several hundred years. The
predominant component of edible konnyaku is a glucomannan called konjac
mannan (KJM). Edible konnyaku is made from the konjac flour, which is
obtained from the dried tuber of this plant. KJM flour is obtained by grinding
the tuber root of the Amorphophallus Konjac C. Koch. plant and is
traditionally
used as a food and remedy in the Far East. In addition to previous findings
(Vuksan et al. (1999)), other findings have shown it to improve cholesterol
levels (Arvill et al. (1995)), hypertension, and glycemia (Doi et al. (1979);
Shima et al. (1982)).
Konjac-mannan was chosen as the fibre because it represents a
polysaccharide with one of the highest viscosities (Kiriyama et al. (1972)).
The physiologically active component is a high molecular weight
glucomannan polymer, which, when taken as a supplement, has been shown
to have effects in lowering lipids (Arvill et al. (1995); Terasawa et al.
(1979);
Venter et al. (1987)), systolic blood pressure (sBP) (Arvill et al. (1995)),
and
glycemia (Doi et al. (1979); Shima et al. (1982)).
Several techniques are known in the art for separating konjac mannan
from konjac flour. In one, konjac flour is boiled in water, treated with
Fehling's
solution to convert the mannan to its copper complex, and the latter is
decomposed again into the mannan after, purification, as disclosed in J. Agr.
Chem. Soc. Japan, 6, 991-995 (1930). In another, konjac flour is extracted
with water, impurities are removed by precipitating with ethanol and
redissolving the precipitate in water several times, and drying the
precipitate
finally obtained to obtain pure konjac mannan, as disclosed in Bull. Chem.
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Soc. Japan, 49, 298-322 (1927). Water-soluble konjac mannan capable of
undergoing gelation when heated in an aqueous alkaline solution may be
used as described in US 3,973,008. Briefly it is obtained by extracting the
ground tuber of the konjac plant with water, separating insoluble matter,
dialyzing the solids-free liquid against water and then lyophilizing the
dialyzed
liquid to remove water. A person skilled in the art would be familiar with
suitable konjac mannan preparation protocols for use herein. All such
preparation methods are intended to be encompassed within the scope of the
present invention.
The present invention provides an improved konjac mannan
composition.
The konjac mannan composition or mixture of the invention preferably
comprises konjac mannan and a substance that can increase the viscosity of
the konjac mannan by 50 - 250%. Accordingly, in one aspect the present
1s invention provides a composition of matter for reducing blood glucose
comprising a konjac-mannan mixture. In another aspect of the invention, a
sufficient amount of the konjac-mannan mixture, when given to an animal
preferably at an appropriate time, reduces blood glucose in the animal,
preferably postprandial blood glucose. Preferably the konjac-mannan mixture
comprises konjac-mannan and a substance capable of increasing the
viscosity of konjac-mannan from 50% to about 250% of konjac-mannan alone.
More preferably the substance comprises about 5% to about 45% by weight
of one or more such substances, preferably polysaccharides.
According to one embodiment a composition as just mentioned
comprises as the one or more polysaccharides xanthan, carragenan, acetan,
guar, or xyloglucana.
According to another embodiment a composition according to the
invention comprises consitituents with the particle size larger than about
1,000
angstroms. In one aspect such a composition is used in controlling cholesterol
levels, preferably lowering cholesterol levels.
Preferably, compositions according to the invention are formulated into
a liquid, powder or formulated as part of a food. However, in another
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embodiment the compositions of the invention are formulated into pills,
capsules and tablets
According to another aspect the present invention provides a method
for reducing blood glucose in an animal comprising administering to the
animal a sufficient amount of a konjac-mannan mixture at an appropriate time
in order to reduce bood glucose, preferably postprandial blood glucose, in the
animal. Preferably the konjac-mannan mixture comprises konjac-mannan and
a substance capable of increasing the viscosity of konjac-mannan to from
about 50% to about 250% of konjac-mannan alone, preferably the substance
comprises from about 5% to about 45% by weight of one or more
polysaccharides, more preferably the one or more polysaccharides are
selected from the group consisting of xanthan, carragenan, acetan, guar, or
xyloglucana.
According to one embodiment of the method, the particle size of the
constituents of a mixture of the invention are larger than about 1000
angstroms.
According to another embodiment of the invention, a mixture according
to the invention is administered orally in an amount of about 1 to about 4
grams per day, preferably the mixture is administered either prior to a meal
or
during the meal.
According to yet another embodiment of the method of the invention
the administration of a mixture according to the invention is by a liquid, a
powder, or as a part of a food product. In another embodiment administration
is by way of tablet, capsule or pill.
Ginseng
A main ingredient of the ginseng is saponin. As the saponin included in
this ginseng there have been known twelve kinds of ginsenside-Ro, -Ra, -
Rbl, -Rb2, -Rc, -Rd, -Re, -Rf, -Rgl, -Rg2, -Rg3, -Rh. These are the one
(ginsenside-Rbl, -Rb2, -Rc) containing sapogenen and protopanaxadiol, and
the one (ginsenside-Re, Rf, -Rgl, -Rg2) containing sapogenen and
protopanaxatriol. The main saponin in the crude drug is ginsenside-Rbl, -
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Rb2, -Rc, -Rgl. The ginsenside-Ro is the same as chikusetsusaponin V, and
the ginsenside-Rbl is the same as saponin D.
Besides these, the ginseng contains essential oil of 0.05%, .beta.-
elemene, panacene (C15 H24) and panaxynol as polyacetylene
compound and further contains choline, vitamin B complex, fatty acid etc.
There are known to be at least seven different types of ginseng and in
the present disclosure a prefered form is American ginseng. There are many
types of ginseng, American, Asian, and others known in the art, and various
subspecies thereof. Not all have the same compositions or therapeutic
effects, even within the same species. Effectiveness can also depend on the
method of preparation of ginseng composition.
According to another embodiment, the ginseng used in the
compositions and methods of the invention can be any ginseng or extract
thereof, for instance American, Asian, Chinese or other ginseng known in the
art. In another embodiment, the ginseng is preferably American ginseng. In
yet another embodiment of the invention, the ginseng or extract thereof has a
diol/triol ratio of about 1.0, most preferably about 1.5 or greater and more
preferably about 1.90 or greater. In another embodiment, the diol/triol ratio
is
about 2.4 or greater.
According to one embodiment of the method of the invention the
amount of ginseng or extract of ginseng is at least about 1000 mg per
administration. In another embodiment, the amount of ginseng administered is
between 1-9 g, preferably 1-4g and most preferably 1-3 g.
In another embodiment, the invention provides a method for
determining whether a ginseng or extract thereof would be useful in
controlling, modulating or lowering blood glucose, preferably postprandial
blood glucose, by determining the diol/triol ratio of the ginseng or extract
and
selected for one that has a diol/triol ratio of about 1.5 or greater. The
selected
extract can then be optionally tested for such activity using the traditional
tests
and assays described herein or known in the art. In another embodiment, the
selected ginseng can also be tested and selected for insulin, lipid profile,
cholesterol and blood pressure control, modulation or decrease. In yet
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another embodiment the selected ginseng can be tested and selected for
effectiveness in the treatment of diabetes and/or cardiovascular disease.
Ginseng, preferably American Ginseng (Panax quinquefolium L.)
reduces postprandial blood glucose in nondiabetic and people with diabetes.
Preferably at least 1000mg of ginseng, preferably American ginseng is
administered together or before the meal (up to 180 min) to see an effect on
the
postprandial blood glucose responses to a test meal. While in nondiabetic it
is
important to take ginseng before meal, in type 2 diabetes, the effect is seen
irrespective of time of consumption (together with meal or up to 180 min
before
meal).
Ginseng, such as American ginseng with a profile of diol/triol ratio of
greater than about 1.0, most preferably 1.5 or greater and more preferably
1.90 or greater. In another embodiment, the triols/diols ratio is about 2.4 or
greater. In one embodiment the composition may be administered in the same
manner as described above to produce glucose lowering effects. Ginseng with
a particular ginsenosides profile may have an effect on increased secretion of
the first phase insulin, similar to conventional diabetic drugs.
Ginseng roots was selected by chemical composition analysis using
HPLC. If ginseng extract is used, it can be prepared by usaly extraction
procedure, by using water to alcohol solution in ratio between 40% to 80% of
water, with the rest being alcohol. Other ginseng, and extract preparation
procedures are known in the art.
The present invention shows for the first time, that ginseng (root
or extract) with protopanaxadiols to protopanaxa triols ratio higher than
about
1.0, most preferable 1.5, lowers the postprandial glycemic response in both
healthy and diabetic individuals and in the long term reduces serum lipids and
blood pressure and thus improves diabetes control. Not intending to be bound
by a particular theory, studies conducted by the present inventors indicate
that
ginseng may potentate insulin secretion potentially through a modulation of
autocoid metabolism linked to nitric oxide production. However, gut mediated
hormone effects may play part in the mechanism of action of AM but this area
has not yet been explored.
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Konjac Mannan and Ginseng
The invention also comprises the administration of both ginseng (such
as American Ginseng (Panax quinquefolium L.) and konjac mannan, jointly. A
combination of selected ginseng, such as American Ginseng (Panax
quinquefolium L.) and Konjac-Mannan fiber (Amorpophallus Konjac k) act
jointly to reduce postprandial blood glucose in people with Type 2 diabetes
more then each individual material. The efficacy is not attainable by either
treatment. The ginseng can be any ginseng or formulation thereof, preferably
as described above, more preferably that has a diol/triol ratio of greater
than
about 1.5 or more. The konjac-mannan is preferably the konjac mannan mix
as described above. The konjac mannan and ginseng can be prepared and
administered in separate compositions or can be formulated into one
composition.
While not wishing to be bound by any particular theory, the possible
mechanism of action for the hypoglycaemic effect of konjac/ginseng is to
increase insulin secretion/sensitivity, and together with konjac mannan's
ability
to slow nutrient absorption and also improve insulin sensitivity these
combined
effects result in lower prolonged elevation of postprandial blood glucose and
have applications in prevention and treatment of diabetes and heart disease.
Thus there are two products, operating through different pathways, that
significantly reduce key risk factors for diabetes and cardiovascular disease.
Compositions
For the purposes of administration by means other than incorporation
within a food, the compositions described herein can be prepared by per se
known methods for the preparation of pharmaceutically acceptable
compositions which can be administered to subjects, such that an effective
quantity of the composition of the invention is combined in a mixture with a
pharmaceutically acceptable vehicle, thereby allowing for the administration
of
a sufficient amount of the composition. Suitable vehicles are described, for
example, in Remington's Pharmaceutical Sciences (Remington's
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Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., USA
1985). On this basis, the compositions include, albeit not exclusively,
solutions of the substances in association with one or more pharmaceutically
acceptable vehicles or diluents, and contained in buffered solutions with a
suitable pH and iso-osmotic with the physiological fluids.
One would appreciate that the effective or sufficient amount of a
composition of the invention can vary depending on the individual, such as
sex, age, and weight of the individual, and/or severity of a condition, such
as
diabetes. It may also in certain cases vary on the time of administration,
i.e.
post or pre meal and time it takes for the substance to clear or be
metabolized
by the body. It may also vary depending on whether the compositions are
administered separately or with another substance that may modulate the
effectiveness of the composition. For instance the effective amount of konjac
mannan may vary depending on whether it is administered alone or in
1s conjunction with a ginseng or another substance that may act
synergistically
with it. It may also vary depending on the concentration and mode of
preparation of the konjac mannan being administered. The above similarly
applied to the administration of ginseng.
Such compositions in one aspect are administered orally, such as by
liquid, powder or formulated as part of a food, such as a biscuit. It could be
administered in the form of pills , capsules and tablets. Other forms of
administration may also be suitable.
Such composition can be used in the modulation or control, preferably
decrease, of blood glucose, preferably postprandial blood glucose. In another
aspect the compositions of the invention can also be used to modulate or
control insulin levels in an animal, preferably decrease such levels. The
compositions of the invention can also be used to modulate a lipid profile of
a
patient in need thereof. They can also be used to modulate blood pressure,
preferably to lower blood pressure. In a preferred embodiment the invention
can be used in the treatment of animals, preferably humans, in need of
controlling their glucose levels, such as in the management and treatment of
diabetes. In another embodiment, the compositions of the invention can be
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used in the management and treatment of cardiovascular disease and
associated conditions, such as high blood pressure, high cholesterol level. In
another embodiment, the compositions of the invention, especially the
ginseng compositions, can be used to modulate or control nitric oxide levels
in
an animal.
The following non-limiting examples are illustrative of the present
invention:
EXAMPLES
GENERAL METHODS FOR EXAMPLES 1-4
Subjects
Eleven diabetic patients (5 men, 6 women) gave written informed
consent to participate in the present study that was approved by the Human
Ethic Committees of St. Michael's Hospital and the University of Toronto. All
had hyperlipidemia, hypertension, and type 2 diabetes (mean serum C-
peptide 701 351 pmol/L), with a minimum of three years since the onset of
all three conditions. They were taking medications to control each of the
three
risk factors, consuming a National Cholesterol Education Program (NCEP)
Step 2 diet Executive Summary of the Third Report of the National
Cholesterol Education Program (NCEP) Expert Panel on Detection,
Evaluation, and Treatment of High Blood Cholesterol in Adults.
JAMA;285:2486-2497,2001, not smoking, nor taking alcohol, and leading
sedentary lifestyles at recruitment. Two participants had a history of
atherosclerotic heart disease, but none had evidence of recent myocardial
infarction, unstable angina, or congestive heart failure. Exclusion criteria
were
a family history of premature CHD, hypothyroidism, renal, hepatic, or
gastrointestinal disease. Table 1 provides baseline demographic,
anthropometric and clinical characteristics of the study participants.
Study Design
The study employed a double-blind, placebo-controlled, cross-over
design, where all subjects were maintained on the same dosage of their
medications throughout. The study began with an 8 week baseline period
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over which participants followed an NCEP Step 2 ad libitum diet: Executive
Summary of the Third Report of the National Cholesterol Education Program
(NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood
Cholesterol in Adults. JAMA;285:2486-2497,2001, documented by three non-
consecutive days of food records every two weeks. This was followed by the
experimental phase of the study that consisted of two successive 3 week
treatment periods, separated by a two week washout interval over which
another three day food record was obtained. During the first treatment period,
subjects were randomly assigned to either the KJM+ (Step 2 metabolically
controlled diet enriched with fiber) or the control treatment (the same diet
enriched with wheat bran [WBJ fiber). For the second treatment period, the
subjects were crossed-over. The study began with 5 subjects taking the
KJM+ treatment and 6 the control.
Diet
Both treatments consisted of a three-day rotating Step 2 diet with three
meals per day provided under metabolic conditions. All foods were pre-
weighed, packaged and couriered to participants for consumption at home or
at work. The mean macronutrient profile conformed with a Step 2 diet.
Energy intakes for weight maintenance were provided according to Lipid
Research Clinics Tables with adjustment for physical activity (The Lipid
Research Clinics Population Studies Data Book. Vol. 2. The prevalence
study-nutrient intake. Washington DC: Government printing office (NIH
publication no. 82-2014) (1982)). Total dietary fiber was administrated at
2g/412 kJ (100kcal), with a mean daily intake according to energy intake
ranging from 24g to a plateau of 50g for those consuming 2500Kcal per day
or more. The actual diet consumed is presented in Table 2.
The two treatments differed only in the type of fiber. On the KJM+
treatment, participants received KJM+ enriched biscuits, whereas on the
control treatment they received an equal quantity of wheat bran (placebo)
biscuits. Subjects were instructed to take biscuits together with an 8oz
beverage, 3 times daily as a snack, including once at bed-time. Both were
produced and provided by Dicofarm S.p.A, Roma, Italy, (commercially
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available in Italy as "Dicoman " biscuits). They had similar nutrient profiles
(Table 2) and were indistinguishable in taste and appearance. KJM+ biscuits
contained approximately 15% KJM flour of which 69% was the active high
viscosity glucomannan, 15% other polysaccharides, and 16% excipients by
weight. The KJM flour mix used in the biscuits comprised in % by weight:
69% KJM, 21% xanthan and 10% caragenan. Because KJM flour comprised
half (1g/412 kJ [100 kcal]) of the total fibre on the KJM+ treatment,
approximately 0.7g/412 kJ (100 kcal) was glucomannan. Wheat bran biscuits,
in contrast, had a lower proportion of fiber than KJM+ biscuits (Table 3).
Approximately 14g/day of wheat bran fibre derived from standardized
American Association of Cereal Chemist hard red wheat bran was, therefore,
added to the control (WB) diet to compensate for these fiber differences.
Any food from the metabolic diet together with study biscuits not
consumed was brought to the clinic for weighing to measure compliance.
Dietary changes found to occur during the first three-week treatment period
were duplicated prior to food delivery for the second treatment period for
each
participant.
Laboratory Methods
Serum blood samples were immediately separated and stored in four
aliquots at -70 C after collection. They were thawed at the end of the study
for analysis of total cholesterol, HDL, and triglycerides (TRIG) measured
enzymatically (McNamara et al. (1987); Warnick et al. (1982)). LDL content
was estimated by the formula of Friedewald et al. (Friedewald et al. (1972)).
Apolipoprotein (Apo) Al and B were determined by rocket
immunoelectrophoresis (Fruchart et al. (1982)). Fasting blood glucose was
analyzed by a hexokinase method using a Cobas Mira Autoanalyzer (Roche
Diagnostic, Mississauga, Canada). Serum fructosamine was analyzed in
triplicate using the Cobas Fara 11 (Lloyd et al. (1984)) and plasma insulin in
duplicate by radioimmunoassay with reagent from ICN Biomedicals, Inc.
(Horsham, Pennsylvania) (Livesey et al. (1980)). Finally, C-peptide was
determined by radioimmunoassay (Kuzuya et al. (1976)).
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Physical measurements were obtained by standard techniques. Blood
pressure was taken and expressed as the mean of three measurements to
the nearest 2 mm Hg on both arms. Fasting body weight was determined
using a beam scale in light clothing, with an emptied bladder and in bare
feet.
Waist and hip circumferences were measured by soft non-stretchable tape on
the narrowest and widest parts of the trunk.
Energy and nutrient analysis of the diets was calculated using US
Department of Agriculture data (The Agriculture Research Services.
Composition of Foods, Agriculture Handbook No. 8. Washington, DC, US
Department of Agriculture, 1992). The nutrient composition of the treatment
biscuits was analyzed, using the Prosky method to determine fiber content
(Prosky et al. (1985)
Statistical Analyses
Results are expressed as mean SEM, except for age, anthropometric
measurements and nutrient intake (mean SD). Data were analyzed by the
Statistical Analysis System (SAS) (SAS Institute Inc.: SAS/STAT User's
guide. Version 6, 4 th ed. Cary NC: SAS Institute Inc. 1989)). Differences in
serum lipids, apolipoproteins, glycemia, blood pressure and body weight
between the beginning (week-0) and end (week-3) of each treatment (control
and KJM+) were assessed by two-tailed Student's t-test for paired data
(PROC UNIVARIATE). Analysis of covariance (ANCOVA) with the facility of
General Linear Model procedure (PROC GLM) was used to test for
differences in these same parameters between the two treatments.
Adjustment for multiple comparisons was made by the Bonferroni-Hochberg
procedure (Hochberg (1988)) for primary (fructosamine, total/HDL cholesterol
ratio, and sBP) and secondary (body weight; total, LDL and HDL cholesterol;
Apo A-1; Apo B; Apo A-1/B ratio; glucose; insulin; and dBP) endpoints
separately. P-values for each endpoint were ordered sequentially and
contrasted with the corresponding adjusted comparison wise critical alpha (a)
levels. Null hypotheses were rejected only if the p-values were less than
their
corresponding a-values (Hochberg (1988)). Control of individual variation from
the repeat measures aspect of the design was addressed by incorporating the
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random subject effect as well as the starting measurement. Diet, sex, and
phase effects were also incorporated in this model. To test for confounding
effects of body weight on study parameters, Pearson correlations were
performed (PROC CORR procedure).
EXAMPLE 1
All participants followed the experimental protocol with little difficulty.
According to three-day food records collected over the baseline and washout
periods, subjects ate their usual low-fat (<25% energy) and high-fiber (>27g
per day) diets (Table 3). In addition, during the treatment periods, returned
food and biscuits from metabolic diets indicated that subjects consumed an
average of 93% and 95% of diet calories prescribed on the KJM+ and control
(WB) treatments respectively and 88% (137g/day) KJM+ test and 91%
(142g/day) WB placebo biscuits. Consumption patterns translated into an
insignificant decrease in body weight during both treatment periods (Table 4).
There was no correlation between changes in weight and serum lipids, glucose
or blood pressure (data not shown). The only side effect experienced was a
transient complaint of flatulence and soft stools reported by 37% and 24% of
participants during the KJM+ and the control (WB) treatments respectively, but
none refused to continue the study.
EXAMPLE 2
Blood lipids were improved during KJM+ treatment compared to control
(Table 4). The primary lipid endpoint, total/HDL cholesterol, decreased
significantly by 5.7 2.3% (P=0.034, a=0.05) during the KJM+ treatment
compared to an insignificant increase of 4.7 4.4% (P=0.316, (X=0.017) on
control. The resultant between-treatment decrease of 10 4.0% on the KJM+
treatment was significant (P=0.028, (x=0.05). The secondary endpoints of total
and LDL cholesterol also fell significantly by 16 2.7% (P=0.001, a=0.005),
25 3.9% (P=0.001, a=0.005), during KJM+ treatment compared to 4.9 3.7%
(P=0.20, (x=0.006), and 4.8 5.9% (P=0.45, (x=0.008) on control. Resultant
between-treatment differences of 11 4.2% (P=0.025, a=0.005) and 19 6.8%
(P=0.033, a=0.006), were insignificant, however, after correction by the
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Bonferroni-Hochberg procedure. The combined fall in total cholesterol and LDL
on the KJM+ treatment indicated reclassification of the lipid status of 6 of
the
11 subjects from elevated to normal cholesterolemia (<5.2mmol/L) (2). Values
for LDL, however, were derived from only 9 subjects, because two of the 11
participants had serum triglycerides levels over 4.5 mmo/L, not allowing for
calculation by the Friedewald equation.
Similar results were observed for Apo B and the Apo B/A-1 ratio. During
KJM+ treatment both fell significantly by 14 3.4% (P=0.002, a=0.006) and
8.6 2.3% (P=0.004, a=0.007), compared to 3.0 5.0% (P=0.57, (X=0.013) and
3.0 4.8 % (P=0.55, a=0.01) on control respectively. These changes, however,
resulted in an insignificant between-treatment difference of 11 4.3% (P=0.025,
(x=0.005) and 5.6 4.5% (P=0.24, a=0.008), after correction by the Bonferroni-
Hochberg procedure.
In contrast, such effects were not seen on HDL, Apo A-1, or triglycerides.
During KJM+ and control treatments, HDL and Apo A-1 decreased
insignificantly, for insignificant between treatment changes. Similarly,
during
both treatments, triglycerides increased insignificantly, with no significant
difference between treatments.
EXAMPLE 3
Improvements in glycemic control were observed on the KJM+ treatment
compared to control (Table 4). The primary glycemic endpoint, serum
fructosamine, was reduced insignificantly during both the KJM+ and control
treatments by 6.1 2.4% (P=0.03, a=0.025), and 0.5 1.4% (P=0.751, a=0.05)
respectively, after correction by the Bonferroni procedure. The resultant
between treatment decrease of 5.7 1.7% on KJM+ was nevertheless
significant (P=0.007, a=0.017). No significant differences between treatment
regimes were seen for the secondary endpoints of insulin or glucose, although
during the KJM+ treatment, fasting glycemia fell significantly by 11 3.0%
(P=0.004, a=0.008) compared to 1.5 6.1% (P=0.804, a=0.013) on control.
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EXAMPLE 4
An improvement in blood pressure was also observed on the KJM+
treatment compared to control (Table 4). The primary blood pressure
endpoint, sBP, decreased significantly on KJM+ supplementation by 5.5 1.4%
(P=0.003, (x=0.017), compared to 1.4 2.7% (P=0.62, (x=0.03) on control,
producing a significant between-treatment difference of 6.9 2.5% (P=0.021,
a=0.025) or 9.4 3 mm Hg. During both treatments, diastolic blood pressure
(dBP), however, remained virtually unchanged with no significant difference
between treatments. The result was a reclassification in sBP status from
moderately high to normotensive (<135mmHg) in 5 of 11 subjects after the
KJM+ treatment.
DISCUSSION OF EXAMPLES 1-4
Examples 1-4 illustrate that the addition of 0.7g/412kJ (100kcal) of high
viscosity glucomannan (KJM mix) in biscuit form to conventional CHD
treatment (a low saturated fat diet combined with drug therapy) improved
metabolic control beyond the effect of conventional treatment alone in high-
risk individuals with type 2 diabetes. Amelioration in three major CHD risk
factors - hyperglycemia, hypertension, and hyperlipidemia - relative to a
matched placebo control treatment as measured by the primary endpoints
fructosamine, sBP, and total/HDL cholesterol respectively was observed.
Differences between secondary glycemic, blood pressure, and lipid endpoints
were insignificant after adjustment for multiple comparisons by the Bonferroni-
Hochberg procedure. With greater power derived from a larger sample size,
significance might have been achieved in these cases.
To achieve similar metabolic benefits, the recent dietary
recommendations of the American Diabetes Association have changed their
emphasis from encouraging carbohydrate and less processed fiber foods to
increased consumption of monounsaturated fat (American Diabetes
Association (ADA): Nutrition Recommendations and principles for people with
diabetes mellitus. Diabetes care 22:S42-S43, 1999). Their reasoning is that
fiber has only very modest effects on LDL cholesterol and does nothing to
raise HDL cholesterol levels. Nevertheless, the diet usually prescribed for
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the management of CHD risk factors in people with diabetes resembles an
NCEP Step 1 or 2 diet. The recommendations for these diets are as follows:
for Step 1, of total calories <30% from total, <10% from saturated, and <10%
from polyunsaturated, with <300mg/day of cholesterol and for Step 2 the
same except <7% of calories from saturated fat with <200mg/day of
cholesterol. In the two well-controlled clinical studies in this area,
limitations
of the diets are evident. Hunninghake et al., following hypercholesterolemic
subjects on a Step 2 diet for three months, found that LDL was reduced by
only 5% (Hunninghake et al. (1993)). Schaefer and colleagues found a
reduction in LDL in subjects provided a Step 2 diet on a metabolic basis to be
as much as 17%, but with adverse effects on other lipid parameters and no
effect on total/HDL cholesterol ratio (Schaefer et al. (1995)). A high inter-
subject variability in LDL reductions was also noticed. The present results,
however, showed that KJM+ treatment resulted in an improvement in lipid
ratios. The suggestion is that a Step 2 diet supplemented with KJM+ may
confer additional benefits over this diet alone.
Lipids
Improvements in blood lipid control have previously been shown when
Step 2 diets were supplemented with soluble fibre from different dietary
sources (Jenkins et al. (1993)) or fibre supplements (Anderson et al. (1986);
Olson et al. (1997)). While such studies have reported reduced total and LDL
concentrations, few, as has been the case for NCEP diets, have reported
improved lipoprotein ratios. Out of the three lipid trials that used KJM+
(Arvill
et al. (1995); Terasawa et al. (1979); Venter et al. (1987)) the former two
did
not show a significant change in these ratios. In contrast, Venter and
coworkers (Venter et al. (1987)) found 4.5g/day glucomanan significantly
improved both LDL and the LDL/HDL ratio in 18 hypercholesterolemic
subjects. The present examples showed a more significant 10 4.0% decrease
in the total/HDL ratio were noticed on the KJM+ treatment compared to
control. The mechanism by which the KJM+ supplemented biscuits had this
lipid lowering effect is not clear. While not wishing to be bound by any
particular theory, possibilities include an inhibition of cholesterol
absorption in
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the ' jejunum (Ebihara et al. (1989)) and bile acid absorption in the ileum
(Kiriyama et at. (1974)) or less postprandial stimulation of HMG CoA
reductase (Jenkins et at. (1993)). Other options include the generation of
short chain fatty acids by colonic microflora, predominantly propionate, which
may decrease hepatic cholesterol synthesis (Venter et al. (1990)).
Glycemic control
Improvements in diabetes control after soluble fibre supplementation
have been shown (Morgan et al. (1990)). KJM, in particular, has been shown
to have a beneficial effect following both acute (Shima et al. (1982)) and
long-
term (Doi et al. (1979); Shima et at. (1982)) administration. In the present
invention KJM+ treatment compared to control, a 5.7 1.7% reduction was
observed in serum fructosamine, a short-term marker of diabetes control, with
no effect on either fasting glucose or insulin concentrations. These results
were not altered by excluding four subjects treated with insulin. An effect of
the gel forming KJM+ on digestion may explain this finding. It has been
suggested that decreases in glucose and insulin levels after the consumption
of water-soluble fibers are related to slower rates of food absorption in the
small intestine associated with increased viscosity (Ebihara et al. (1981)).
KJM has been shown to have very high viscosity, approximately five times
higher than guar gum (Ebihara et al. (1981)) and considerably more than
pectin (Venter et at. (1987)). Consequently, in some studies it has been given
at half the dosage relative to these other fibers (Ebihara et al. (1981)). The
present inventors have further increased the viscosity of KJM by the addition
of polysaccharides such xanthan, caragenan, acetan, guar or xyloglucena.
Blood pressure
Finally, although few studies have demonstrated an effect of fibre on
blood pressure, significant reductions both in sBP and dBP have been
reported after consumption of guar granulates (Landin et al. (1992)) and
soluble dietary fibre supplements (Alison et at. (1992)). The same effect has
been shown for KJM, but only on sBP (Arvill et al. (1995)). This last finding
agrees with the results set out in the present examples, in which KJM
treatment significantly reduced sBP by 6.9% compared to control but did not
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affect dPB. The commonly recommended oat bran, in contrast, has been
shown to affect neither systolic nor diastolic blood pressure (Swain et al.
(1990)). While not wishing to be bound by any particular theory, a possible
mechanism for the blood pressure lowering effect of soluble fibers may
involve increased insulin sensitivity (Anderson et at. (1986)), which may
reduce blood pressure by influencing sodium absorption in the distal tubule,
increasing sympathetic nervous system activity and peripheral vascular
resistance (Modan et al. (1985)).
The effect of KJM+ fibre supplements on the three CHD risk factors
persist even in subjects who are taking conventional drug therapy
concurrently. Consistent with the findings set forth in the examples, a
combination of fiber and drugs has been shown to be more effective clinically
than the drug given alone in improving metabolic control. Toumilehto and
coworkers (Tuomilehto et al. (1989)) found that the viscous soluble fiber guar
gum and gemfibrozil administered together reduced total cholesterol and
LDL/HDL ratio significantly more than gemfibrozil and placebo. Elsewhere
this same effect has been noticed for blood glucose and blood pressure. A
significant reduction was found in postprandial blood glucose after
consumption of sulfonylyurea (glibenclamide) and glucomannan with a test
meal compared to sulfonylyurea alone with the same test meal (Shima et al.
(1983)). Similarly, a significant decrease in diastolic blood pressure was
noticed after administration of guar gum compared to placebo in patients
receiving drug treatment for hypertension (Uuistupa et al. (1984)). Together
these findings suggest that highly viscous soluble fibre may augment or
potentiate the effect of drugs.
In conclusion, the application of KJM+ supplementation in the high-risk
diabetic study group of the examples demonstrated simultaneous
improvement in all three diet-modifiable risk factors, indicating a reduction
in
overall CHD risk (Jenkins et al. (1995)). One of the benefits is that KJM+
supplemented therapy may lower required drug dosages and improve overall
cost-effectiveness and acceptability of treatment. Although it is agreed that
food should be the normal way to achieve an adequate fiber intake, fiber
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supplemented foods have advantages in the treatment of individuals at high
risk for CHD and represent a possible intermediate step between diet and
drug therapy.
GENERAL METHODS FOR EXAMPLES 5-8
Subjects
278 free-living subjects were screened from the Canadian-Maltese
Diabetes Study between the age of 45 and 65 years. This population is
known to have one of the highest rates of diabetes (Katona et al. (1983)).
Thirty eight of them satisfied the initial inclusion criteria: impaired
glucose
tolerance (IGT) (World Health Organization Diabetes Mellitus: Report of the
World Health Organization Study Group. Technical report No. 727:9-15,
1985); clinical absence of CHD; body-mass index of less than 30 kg/m2; not
taking medications for hyperglycemia, hyperlipidemia or hypertension; not
smoking; nor consuming more than two alcoholic drinks per day. These
is subjects were further screened for the presence of the full multiple
metabolic
syndrome (Trevisan et al. (1998)) (syndrome X). This included moderate
hypertension (>135/85 and less than 145/95 mm Hg), dyslipidemia (low-HDL
[levels below 0.9 mmol/L for men and 1.2 mmol/L for women], and elevated
triglycerides [greater than 2.3 mmol/L and less than 4.5 mmol/L]). Based on
power analysis from the previous study (Vuksan et al. (1999)) and Example 1,
eleven subjects (5 men, 6 women) who qualified were recruited. In addition to
meeting the above criteria, their fasting (98 13pmol/L) and 2-hour
postprandial (439 68pmol/L) plasma insulin levels was greater (p<0.05) than
two standard deviation of the initial screening pool (71 8 and 316 47pmol/L
respectively). All eleven also had moderately high serum cholesterol
(between 5.2 and 6.7 mmol/L) and were sedentary, with a mean ( SD) age of
55 4 years (range: 46-61 years); a BMI of 28 3kg/m2; a waist-hip ratio of
0.98 0.2 (waist: 96 12cm) in men and 0.91 0.4 (waist: 87 19cm) in women.
They gave written informed consent to participate in the current study that
was approved by the Human Ethic Committees of St. Michael's Hospital and
the University of Toronto.
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Study Design
The study employed a double-blind, placebo-controlled, cross-over
design that was identical to that used in the previous study (Vuksan et al.
(1999)) and Example 1. It began with an 8-week baseline period during which
participants followed a National Cholesterol Education Program (NCEP) Step
2 (American Diabetes Association (ADA): Nutrition Recommendations and
principles for people with diabetes mellitus. Diabetes Care 22:s42-s43,
1999)) ad libitum diet, documented by three non-consecutive days of food
records every two weeks. This run-in phase was included to eliminate
io possible effects of dietary change on metabolic parameters. The
experimental phase of the study followed. This consisted of two successive
3-week treatment periods, separated by a two-week washout interval over
which a Step 2 diet was followed and documented by another three-day food
record. During the first treatment period, subjects were randomly assigned to
either the KJM+ (Step 2 metabolically controlled diet enriched with KJM+
fiber) or the control treatment (the same diet enriched with wheat bran [WB]
fiber). For the second treatment period, the subjects were crossed-over.
Blood collection, weight, blood pressure, and waist and hip measurements
were done at the beginning and end of each 3-week treatment period. The
study began with 5 subjects taking the KJM+ treatment and 6 the control.
Diet
Both treatments consisted of a three-day rotating Step 2 diet with three
meals per day provided under metabolic conditions. All foods were pre-
weighed, packaged and couriered to participants for consumption at home or
at work. The mean macronutrient profile closely conformed to a Step 2 diet (of
calories <30% from total fat, <7% from saturated fat, and <300mg/day
cholesterol) (American Diabetes Association (ADA): Nutrition
Recommendations and principles for people with diabetes mellitus. Diabetes
Care 22:s42-s43, 1999)). Energy intakes for weight maintenance were
provided according to Lipid Research Clinics Tables with adjustment for
physical activity (24). Total dietary fiber was administrated at 1.5g/100kcal,
with a mean daily intake according to energy intake ranging from 24g to a
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plateau of 40g for those consuming 2800kcal per day or more. The actual diet
consumed is presented in Table 5.
The two treatments differed only in the type of fiber. On the KJM+
treatment, participants received KJM+ enriched test biscuits whereas on the
WB-control treatment they received an equal quantity of wheat bran control
biscuits. Subjects were instructed to eat an equal amount of biscuits together
with an 8oz beverage, 3 times daily as a snack, including once at bedtime.
Both were provided by Dicofarm S.p.A, Roma, Italy. They had similar nutrient
profiles and were indistinguishable in taste and appearance. KJM+ biscuits
contained approximately 10% KJM flour of which 69% was the active high
viscosity glucomannan, 15% other polysaccharides, and 16% excipients by
weight (Vuksan et al. (1999)). The KJM flour mixture of the KJM+ biscuits
comprised 69% + 17% xanthan, 9% caragenan, and 8% guar. Because
KJM flour comprised half (0.75g/100 kcal) of the total fibre on the KJM+
treatment, approximately 0.5g/100 kcal (8-13g/day) was glucomannan.
Wheat bran biscuits, in contrast, had a lower proportion of total dietary
fiber
than KJM+ biscuits. Approximately 11g/day of wheat bran fibre derived from
standardized American Association of Cereal Chemist hard red wheat bran
was, therefore, added to the WB-control diet to compensate for these fiber
differences. Subjects were instructed to sprinkle the additional fiber on
cereal,
yogurt, and/or other compatible foods to improve palatability.
Any foods from the metabolic diet together with study biscuits not
consumed during the study were returned to the clinic for weighing to
measure compliance. Dietary changes found to occur during the first three-
week treatment period were duplicated in the diets for the second treatment
period for each participant.
Laboratory Methods
Laboratory methods were identical to those used in (Vuksan et al.
(1999)). In brief, blood samples were separated immediately and stored as
serum in four aliquots at -70 C after collection. They were thawed at the end
of the study for analysis of total cholesterol, HDL, and triglycerides
measured
enzymatically. LDL content was estimated by the formula of Friedewald et al.
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Apolipoprotein (Apo) Al and B were determined by rocket
immunoelectrophoresis. Fasting blood glucose was analyzed by a hexokinase
method using a Cobas Mira Autoanalyzer (Roche Diagnostic, Mississauga,
Canada). Serum fructosamine was analyzed in triplicate using Cobas Fara II
and plasma insulin in duplicate by radioimmunoassay with reagent from ICN
Biomedicals, Inc. (Horsham, Pennsylvania). C-peptide was determined by
radioimmunoassay.
Statistical Analyses
Results are expressed as mean SEM, except for age, anthropometric
measurements and nutrient intake (mean SD). Data were analyzed by the
Statistical Analysis System (SAS Institute, Cary, NC). Differences between
the diets were assessed by two-tailed Student's t-test for paired data
(univariate procedure). This same statistic also assessed differences in
serum lipids, apolipoproteins, glycemia, blood pressure and body weight
between the beginning (week-0) and end (week-3) of each treatment (WB-
control and KJM+). Analysis of covariance (ANCOVA) with the facility of
General Linear Model (GLM) procedure was used to test for differences in
these same parameters between the two treatments. Control of individual
variation from the repeat measures aspect of the design was addressed by
incorporating the random subject effect as well as the starting measurement.
Diet, sex, and phase effects were also incorporated in this model. Adjustment
for multiple comparisons was made by the Bonferroni-Hochberg procedure
(Hochberg (1988)). P-values for each endpoint were ordered sequentially and
contrasted with the corresponding adjusted comparison-wise critical alpha (a)
levels. The null hypotheses were rejected only if p-values were less than
their
corresponding a-values.
EXAMPLE 5
All participants followed the experimental protocol with little difficulty.
Returned food from metabolic diets indicated that subjects consumed an
average of 96% and 95% of diet calories prescribed on the KJM+ and WB-
control treatments respectively. Returned biscuits indicated they consumed
81% (97g/day) of KJM+ and 86% (103g/day) of WB-control biscuits.
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Consumption patterns translated into an insignificant decrease in body weight
during both treatment periods with no difference between treatments (Table
6). The only side effect experienced was a transient complaint of flatulence
and soft stools reported by 3 and 2 of the participants during the KJM+ and
the WB-control treatments respectively, but none chose to discontinue the
study.
EXAMPLE 6
Blood lipids improved during KJM+ treatment compared to WB-control
(Table 6). Total and LDL cholesterol fell significantly by 19 2.7% (P<0.0001)
and 29 3.4% (P<0.0001) during KJM+ treatment compared to 6.3 3.4%
(P=0.088) and 6.6 5.0% (P=0.231) on control. The between-treatment
differences were 12.4 3.1% (P<0.005) and 22 3.9% (P<0.003) respectively.
The combined fall in total cholesterol from 6.2 0.3 to 5.0 0.2mmol/L and LDL
from 3.9 0.2 to 2.8 0.2mmol/L on KJM+ treatment indicated reclassification of
1s the lipid status of the group (8 of 11 subjects) from elevated to normal
cholesterolemia (National Cholesterol Education Program: Second report of
the expert panel on detection, evaluation, and treatment of high blood
cholesterol in adults (adult treatment panel II). Circulation 89:1333-1445,
1994)). Similar results were observed for Apo B. During KJM+ treatment Apo
B fell significantly by 19 2.8% (P<0.0004) compared to 4.5 4.5% (P=0.34) on
control, for a significant difference of 15.1 4.3% (P<0.0004) between the
treatments.
In contrast, such effects were not seen on Apo A-1, or triglycerides.
During KJM+ and control treatments, HDL decreased significantly on both
treatments (8.5 2.2%, P<0.04 on KJM+ diet and 9.6 2.2%, P<0.003 on WB-
control, with an insignificant between-treatment change (P=0.98). Similarly,
during both treatments, triglycerides increased insignificantly, with no
significant difference between treatments.
Despite this lack of effect of KJM+ treatment on HDL, Apo A-1, or
triglycerides, the decreases in total cholesterol and Apo B were sufficient to
improve lipid ratios. During KJM+ treatment total/HDL, LDL/HDL and Apo B/A-
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1 ratios decreased by 11 3.0% (P<0.005), 22 3.7% (P<0.0002) and 13 3.0%
(P<0.003) respectively. This compares to an insignificant increase of
4.1 4.1% in total/HDL ratio, 0.2 6.3% in LDL/HDL ratio and 0.7 3.6 % in Apo
B/A-1 on WB-control. The resultant between-treatment differences were
15.2 3.4% (P<0.003) for total/HDL cholesterol, 22.2 4.1% (P<0.002) for
LDL/HDL cholesterol, and 13.1 3.4% (P<0.0003) for Apo B/ A-1.
EXAMPLE 7
An improvement in glycemic control was observed on the KJM+ treatment
compared to WB-control (Table 6). Serum fructosamine was reduced during
the KJM+ treatment by 5.6 1.5% (P<0.003), compared to 0.39 1.3% (P=0.77)
on control, with a between-treatment difference of 5.2 1.4% (P<0.002). No
significant between-treatment differences were seen for insulin or glucose
concentrations. On KJM+ however, fasting glycemia fell by 13 2.5%
(P<0.0001) compared to 9.6 4.3% (P<0.05) on control.
EXAMPLE 8
No change in systolic or diastolic blood pressure was observed on
either treatment or between treatments (Table 6).
All above results remained unchanged after adjustment for multiple
comparisons by the Bonferroni-Hochberg procedure.
DISCUSSION OF EXAMPLES 5-8
These Examples demonstrate that the addition of 0.5g/100kcal (8-
13g/day) of high viscosity glucomannan in biscuit form to a high-
carbohydrate/low-saturated fat NCEP Step 2 diet improved metabolic control
beyond diet alone in individuals with the insulin resistance-dyslipidemic
syndrome. Significant reductions in hyperglycemia as measured by the short-
term marker of glycemic control, fructosamine, were observed. Also observed
were significant reductions in hyperlipidemia as measured by total, LDL,
LDL/HDL and total/HDL cholesterol, apo B and apo B/A-1, relative to a
matched WB-control treatment. These findings represent the first to
demonstrate such improvements using soluble fiber in individuals with this
particular cluster of risk factors that also includes the intermediate
diabetic
classification, IGT. [syndrome X]
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Because of the strong implications of this syndrome, a more
aggressive approach has been suggested to achieve similar reductions.
Diabetes and heart disease share common precursors for the development of
atheroslerosis that often co-occur. Long before diabetes becomes manifest,
the,clustering of metabolic abnormalities exerts a synergistic effect on the
atherosclerotic process (Haffner et al. (1990)). Based on findings from
Trevisea and colleagues, cardiovascular disease (CVD) risk appears to
increase linearly with an increase in the number of these risk factors. It is
recommended therefore that insulin resistant patients have their CHD risk
factors managed as if they have established coronary heart disease (Haffner
eta[. (1998)).
Low-fat/high carbohydrate diets may still have promise as a therapeutic
approach. Although there has been a shift away from their advocacy in favor
of those rich in monounsaturated fat (American Diabetes Association (ADA):
Nutrition Recommendations and principles for people with diabetes mellitus.
Diabetes Care 22:s42-s43, 1999), these diets supplemented with fiber may
have similar metabolic advantages. Guar gum, pectin, oat products, and
psyllium added to high carbohydrate diets have been shown to improve total
and LDL cholesterol significantly, with no improvement to triglycerides and
slight or no adverse effects on HDL (Jenkins et al. (1978)). Both guar (Aro et
al. (1981)) and KJM+ (Arvill et al. (1995); Vuksan et al. (1999))
supplementation have also been shown to improve other risk factors,
including glycemia and blood pressure. This lead to support for the use of
guar in the treatment of the multiple metabolic syndrome (Landin et al.
(1992)). Evidence further suggests that supplementation with these soluble
fibers may augment concurrent drug therapy. Improvements in these assorted
risk factors following supplementation have been noticed beyond what was
achieved by drugs alone in subjects receiving hypolipidemic (Aro et al.
(1981);
Vuksan et al. (1999); Tuomilehto et al. (1989)), hypoglycemic (Aro et al.
(1981); Vuksan et al. (1999); Shima et a). (1983)), and hypotensive (Vuksan
et al. (1999); Uuistupa et al. (1984)) medications.
The ability of soluble fiber to improve a high carbohydrate/low fat diet is
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supported by the findings of these Examples. Total and LDL cholesterol were
decreased and glycemic control was improved significantly. Also, although
HDL, Apo A-1, and triglycerides were unaffected, as has been noticed with
other fibers, this was balanced by the significant improvements in the other
lipid endpoints, leading to significant reductions in all three lipid ratios:
Total/HDL, LDL/HDL, and Apo B/A-1. Similar improvements in these ratios
have rarely been reported using dietary interventions (Tuomilehto et al.
(1989); Shima et al. (1983)). Overall, the present findings indicate that a
Step-2 diet supplemented with KJM+ may confer additional benefits over the
Step-2 diet alone, benefits that may be comparable to strategies using
monounsaturated fat.
KJM+ may be better suited than the other major soluble fibers in
improving outcomes with high-carbohydrate/low-fat diets. Although meta-
analyses use variance adjusted values that tend to underestimate
effectiveness, KJM+ can be compared to other soluble fibers in terms of its
lipid lowering ability per gram of fiber, using recent meta-analytical data
(Brown et al. (1999)). Daily intake of glucommanan from KJM+ on this study
produced an average net change in total and LDL cholesterol of -0.084 and -
0.119mmol/L per gram of fiber respectively. These reductions represent
approximately triple the lipid lowering capacity of psyllium (-0.028 and -
0.029mmol/L respectively), oat products (-0.037 and -0.032mmol/L
respectively), and guar gum (-0.037 and -0.033mmol/L respectively) (Brown et
al. (1999)). In the case of pectin, they represent comparable total
cholesterol
lowering capacity (-0.070 mmol/L) and approximately twice the LDL lowering
capacity (-0.055mmol/L) (Brown et al. (1999)). The very high viscosity of
KJM+ used in this present study may explain these differences. It has been
shown to be approximately five times higher than that of guar gum (Ebihara et
al. (1989)) and beta-glucan (Wood (1990)), and considerably more than that
of pectin (Venter et al. (1987)).
Contributions made by its theological properties may offer insight into
the proposed mechanism by which the KJM+ supplemented biscuits had their
beneficial effects. While not wishing to be bound to any particular theory,
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possibilities for its lipid lowering action may include an inhibition of
cholesterol
absorption in the jejunum (Venter et al. (1987) and bile acid absorption in
the
ileum (Kiriyama et al. (1974)) mediated by viscosity or less postprandial
stimulation of HMG CoA reductase (Jenkins et al. (1993)). Other options
include the generation of short chain fatty acids, predominantly propionate,
by
colonic microflora that may decrease hepatic cholesterol synthesis (Venter et
al. (1990)). The improvement in glycemic control may be attributable to an
effect of the gel forming KJM+ on rate of digestion. It has been suggested
that decreases in glucose and insulin levels after the consumption of water-
soluble fibers are related to slower rates of food absorption in the small
intestine associated with increased viscosity (Jenkins et al. (1978)). This
mechanism may explain why a reduction in serum fructosamine, but did not
concomitant reductions in fasting glycemia and insulinemia were observed:
KJM+ may be exerting its effect mainly postprandially.
In conclusion, the results in these Examples support the role of KJM+
mix as a means for improving high-carbohydrate diets in the amelioration of
the insulin resistance-dyslipidemic syndrome. Improved metabolic control
resulted in the correction of several risk factors that characterize the
syndrome and figure prominently in the etiology of atherosclerotic CHD.
Example 9 - Effect of Various PolySaccharides on Blood Glucose
Ten healthy male (3) and female (6) volunteers (Age=37;
BMI=26.3kg/m2) were randomly assigned to consume either wheat bran
control, Psyllium fiber, xanthan , KJM alone (FMC Co.) or KJM mix
(preparation: KJM 74%, xanthan 26%) on five separate occasions. Each of
the treatments contained 5 grams of above-mentioned ingredients which was
added to 75g available carbohydrate (300 ml) derived from glucose drink
Glucodex solution (Technolab, Quebec) drink. Capillary blood glucose was
taken after 10-12hrs fasting and 15, 30, 45, 60 and 90 minutes after start of
the test meals. Blood glucose was analyzed using YSI 2300 instrument.
Results are presented as area under the glucose curve (Figure 1) and
absolute and incremental blood glucose levels (Figure 2) for individual time
points. The results indicated that the area under the glucose curve for KJM
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mix (39) was significantly lowered (P<0.011) than control treatment (113;
p<0.02), psyllium (100; p<0.032), xanthan (81; p<0.041) and KJM alone (80;
p<0.027). Incremental and absolute glucose levels were significantly reduced
on KJM mix treatment at time 30 (P<0.05) compared to all other treatment.
In summary, figures 1 and 2 provide illustrations of the significant
effects on blood glucose a konjac mannan mixture of the present invention
has over the effects of individual saccharides and konjac mannan alone. The
mixture of konjac mannan comprised konjac mannan and Xanthan although,
other saccharides which can be use include, carragenan, acetan, guar, or
xyloglucana.
Example 10 - Chronic Feeding of Konjac-Mixture and Effect on
Postprandial Glycemia in Insulin Resistance
Atherosclerosis and diabetes have been characterized as postprandial
phenomena. Recent epidemiological analyses demonstrated that diets with a
low glycemic load reduce their incidence. To investigate the ability of KJM
mix to reduce postprandial glycemia in the insulin resistance syndrome
(syndrome X) that underlies these diseases, 12 participants were studied
(age:55 4y, BMI:28 3kg/m2) who satisfied the criteria for the syndrome (IGT,
reduced-HDL, elevated triglycerides and mild-hypertension) following 21 days
of KJM mix supplementation. In a crossover design, participants were
assigned to take a metabolically controlled NCEP Step-2 diet either with
0.7g/100kcal of KJM mix enriched biscuits (as outlined in Examples 5-8), or
matched wheat bran control biscuits over two 21 day treatment periods (see
Figure 3 for composition profiles). Venous blood samples were drawn at 0, 30,
45, 60, 90, 120, and 180 min after a standard breakfast (see Table 7), before
(day-1) and after (day-3) chronic feeding of the KJM mix or wheat bran
biscuits on each treatment period. Plasma glucose [Figure 4A (day-1), Figure
4B (day-3)] and insulin [Figure 5A(day-1) and Figure 5B (day-3)]
concentration profiles were determined and whole body insulin sensitivity was
calculated using both fasting and postprandial values, according to Matsuda
and DeFronzo (Diabetes Care 1999; 22:1462-70). Area under the curves for
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glycemia (-23 .5% versus 0.4 2.3%, P=0.000022)(Figure 4C) and insulinemia
(-40.5 4.5% versus -2.0 2.9%, p=0.000012) (Figure 5C) were significantly
reduced on the KJM mix treatment compared to wheat bran control treament.
These decreases translated into a significant increase in postprandial insulin
sensitivity on KJM compared to control (55.9 9.2% versus 9.7 4.5%,
P=0.00056). Insulin sensitivity index (ISI) was calculated as follows:
Composite Whole Body ISI = 10,000/(FPG x FPI) x (G x I)
Where 10,000 is the constant; CompositeWhole Body ISI is hepatic &
peripheral tissue insulin sensitivity, FPG is fasting plama glucose (mg/dL);
FPI
is fasting plasma insulin ( U/ml); G is mean glucose after glucose challenge;
and I is mean insulin after glucose challenge.
From this it may be concluded that prolonged consumption of KJM
improves glycemic control, as indicated by lower postprandial glycemia and
insulinemia.
Example 11 - Effect of Highly Refined Konjac Mannan And Konjac
Mannan Mix On Postprandial Glycemia In Normal Individuals
To determine the effect, if any, of viscosity of KJM fiber preparations on
KJM activity, the efficacy of the KJM mix compared with highly refined
commercially available KJM RS (Opta Co., US) on lowering postprandial
blood glucose levels was studied.
First viscosity measurements were taken of 2 KJM fibers, commercially
available highly purified KJM RS produced by Opta C. (KJM 3) and the same
KJM mixed with 23% of xanthan (KJM 1). To determine the difference in
viscosity over time, 1.5 g of each KJM sample was mixed with 150 ml of
water. Theological measurements using a Synchro-electric Viscometer (D.W.
Brookfield Ltd., Cooksville, ON) (Shear 0.6/12, spindle E) were taken at 15,
30, 45, 60, 90, 120, 180min and 24h. Relative viscosity was compared
between the samples. The two KJM fibres from Opta Co. (KJM 3) or the KJM
mix (KJM 1) were compared in vivo testing. In vitro analysis indicated that
viscosity of KJM 1(KJM mix) was significantly higher (42,000 cps) compared
to commercially available most purified konjac RS fibre, KJM 3 (16,200 cps).
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For the in vivo study seven healthy male (4) and female (3) volunteers
(Age=43; BMI=24.7kg/m2) were randomly assigned to consume either wheat
bran control, KJM RS (KJM 3) or KJM mixture treatment containing 23 % of
xanthan gum (KJM 1) on three separate occasions. Each of the treatments
contained 3 grams of above-mentioned ingredients which was added to 50g
available carbohydrate (356 ml) derived from vanilla flavored EnsureTM drink
(Ross Abbott Laboratories). Capillary blood glucose was taken fasting (i.e
after a 10 to 12 hrs fast) and 15, 30, 45, 60 and 90 minutes after start of
the
test meals. Blood glucose was analyzed using YSI 2300 instrument. Results
are presented as area under the glucose curve (Figure 6C) and absolute and
incremental blood glucose levels (Figure 6A and 6B) for individual time
points.
The results indicate that the area under the glucose curve for KJM 1 (68.4)
was significantly lowered (by about 44%) (P<0.015) than control treatment
(120.5), and KJM 3 (P<0.034) treatment (95.6) (lowered by about 21%).
There were significant differences between the two KJM fibers on time.
Incremental and absolute glucose levels were significantly reduced on KJM 1
treatment at time 60 (P<0.02) and 90 minutes (P<0.01) as compared to KJM 3
treatment. When compared to the control meal KJM 3 showed no significance
difference at any time points, or for the area under the glucose curve. As
such
it can be concluded that KJM mixed with polysaccharides such as xanthan in
proportion to act synergistically is more effective on postprandial glycemia
than highly refined KJM alone. KJM in mixture with other polysaccharides will
express its efficacy in glycemic control of healthy individuals.
Example 12 - Effect of American ginseng, Konjac Mannaan Mix
and Combination Thereof on Postprandial Blood Pressure in Type 2
Diabetes.
The effect of the proprietary KJM MIX, AG, and their combination
(AG&KJM MIX) on postprandial blood pressure regulation in type 2 diabetic
individuals as compared with wheat bran control was studied. Seventeen
healthy male (10) and female (7) volunteers (Age=64.7; BMI=29.4kg/m2) were
randomly assigned to consume either wheat bran control, AG, KJM mixture
treatment containing 27 % of xanthan gum (KJM MIX), or combination of AG
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and KJM MIX, on four separate occasions. Each of the treatments contained
3 grams of above-mentioned ingredients which was added to 50g available
carbohydrate (356 ml) derived from vanilla flavored EnsureTM drink.
Standardized lunch was served at 240 minutes time after start of breakfast.
Blood pressure was taken -30 min, 0, 30, 60, 120, 240, 360, and 420 minutes
after start of the test meals. Blood pressure was determined using
conventional mercury sphyngomanometar device. Results are presented as
systolic and diastolic blood pressure (See Figure 7). When compared to the
control treatment, systolic blood pressure (SBP) was significantly reduced
after taking KJM MIX (at time30, 60, 120, and 360 minutes); KJM MIX and AG
combined (at 30,60,120, and 240 minutes), whereas AG lowered SBP only at
30 min and 120 min after start of test breakfast. The diastolic blood pressure
was not different between four treatments. Therefore, it can be concluded that
KJM MIX and AG whether taken alone or in combination reduce SBP in type 2
diabetic individuals. Further, the effect of the combination of KJM and AG
seems to be different than each individual component. This indicates that
there is a different mechanism of action which might be beneficial and
superior to wither of the two components alone.
Example 13- Effect of American ginseng, Konjac Mannaan Mix
and Combination Thereof on Postprandial Blood Pressure, Blood
Glucose and Insulin in Type 2 Diabetes.
The effect of the proprietary KJM MIX, AG, and their combination
(AG&KJM MIX) on postprandial blood pressure, blood glucose and insulin
regulation in type 2 diabetic individuals as compared with wheat bran control
was studied.
Seven volunteers with type 2 diabetes were recruited; duration of
diabetes 8.4 4.9yr, HbAlc 6.9 1%, 86.4 15.9 kg. The study utilized a
crossover, double blind design. Four sets of breakfasts and lunches were
administered in random order. Lunch was a standard meal, only the breakfast
meal contained either wheat bran (control) or KJM mix with or without AG.
The test days were randomized for each subject and were scheduled at least
one week apart. The total test day spanned 7 hours. Blood samples were
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taken using an indwelling catheter at -30,0,5,10, 15,30,45,60,90,120,150,180,
240 (lunch), 270, 300, 330, 360, 390 and 420min. Samples will be analyzed
for glucose and insulin.
Clinical blood pressure was measured using conventional mercury
sphygmomanometer according to Joint National Council (JNC) VI criteria at
-30, 0, 30, 60, 120, 240, 360, 420min. Subjects were asked to record their
satiety levels throughout the day using a bipolar scale ranging from -3
(extremely hungry), 0 (neutral), to +3 (uncomfortably full). The palatability
of
the meals was also recorded on a scale from 1 to 10, where 1 was "dislike
extremely", 5 "neutral" and 10 "delicious". At each visit weight was measured
using a beam scale and total body fat was measured with the Futrex 5000,
using infrared technology. Additional samples were taken at the beginning
and end of the study and measured for hemoglobin levels. This measurement
was suggested by the Ethical Board of St Michael's Hospital as there was a
concern regarding possible anemia.
The results indicated that weight and total body fat did not significantly
differ for each intervention. Hemoglobin decreased significantly by 12.2 g/L.
There were no differences between meals in palatability of either breakfast or
lunch. The blood pressure results look promising.
Postprandial blood glucose was significantly lower in all 3 interventions
(KJM mix, AG, and its combination) compared to control wheat bran. The
lowest results are found to be with combination of KJM mix and AG, indicating
an additive effect of mixing the two compounds (Figure 8A and 8b). Insulin
results indicated significantly higher insulin secretion, but lower on KJM mix
and even lower when 2 compounds were combined. (Figure 9)
Overall, the effect of KJM, and AG in the present study confirmed that
they both beneficially affect postprandial glycemia. Postprandial blood
glucose
was lowered by two mechanisms that are different but complementary;
ginseng increase insulin secretion and KJM mix reduces insulin levels
(increases insulin sensitivity). The effect of the combination of KJM and AG
seems to be different than each individual component. This indicates that
there is a different mechanism of action which might be beneficial and
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superior to wither of the two components alone. The fact that blood glucose
was lowest on KJM mix and AG combination with lowest insulin secreted,
indicate that this combination provides the most effective and economic
intervention of all three. Saving insulin (insulin economy) to achieve lowest
blood glucose results would be extremely beneficial in a diabetes therapy.
Example 14 - The Effect of Protopanaxadiol/Protopanaxatriol
Ratios Of Ginseng On Glucose Levels
There are a wide variety of ginsengs and not all, even ones within the
same species will necessarily have the same effect. As such the composition
profiles of various ginsengs and their effect on blood glucose and insulin was
studied.
The effect of ginsengs with different diols/triaols ratios was
investigated. In series of successful clinical studies American ginseng was
used, with composition of protopanaxadiols (Rbi, Rb2, Rc, and Rd) (diols)
relative to protopanaxatriols (Rgi, Re, Rf) (triols), that had a ratio of
above
approximately 1.5 (i.e. Chinese ginseng with 1.91 ratio and American with
1.51 and 2.44 ratio). As such ginseng with a specific ginsenoside profile
significantly decreases glucose. Ginseng profiles with weight ratios lower
then
1.5 were also studied.
In the studies, ginseng was given with a 75g oral glucose tolerance test
(OGTT) and compared the results to a similar study done previously with
ginseng with higher ratios (Figure 10). Normal subjects were studied with 3
different ginsengs; in a first study 12 (gender: 6m: 6f, age: 31 3years, BMI:
28 2kg/m2) with Chinese ginseng with diaols/triols ratio of 0.8; in the second
and third studies, 10 (gender: 4m: 6f, age: 41 6years, BMI: 26.1 0.4kg/m2)
with AG with diaols/triols ratio of 1.2 and 1.37, respectively. A single-
blind,
crossover design was used in which all subjects received control or 6g, 40min
before a 75g-OGTT. Control in the present study consisted of identical
capsules containing cornstarch, whereas in the previous study it was the 75g-
OGTT done alone. Both protocols followed the Canadian Diabetes
Association guidelines for the OGTT, with venous blood samples drawn at
-40, 0, 15, 30, 45, 60, 90, and 120min. Repeated measures analysis of
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variance demonstrated that there was no significant effect of the AG
containing diols/triols ginsenosides with ratios lower then 1.5. on
incremental
change or area under the curve (AUC) for glycemia or insulinemia. That was
in contrast with results where ginseng with ratios above approximately 1.5 had
beneficial effects. One of the explanation for this discrepancy is that the
level
of total or specific active ginsenosides, possibly in dials fraction rather
then
trials fraction of ginseng. These data suggest that the ginsenoside profile of
ginseng, particularly ratio between protopanaxadials to protopanaxatriols of
above 1.5, so this play a role in its effects.
That part of ginseng's profile that gives improvement in human
metabolism seems to be related to ginseng composition, mainly to ratios
between main ginsneosides groups. The ginsenoside content of the 6
different ginseng grinded root and extracts used in the series of clinical
studies was studied and found that reduction of blood glucose, lipids, and
blood pressure; and also increase in insulin secretion, and nitric oxide, and
reduction in oxidative stress had a high proportion of protopanaxadiols (Rbi,
Rb2, Rc, and Rd) relative to protopanaxatriols (Rgi, Re, Rf).
The present study indicates that an optimal level of this ratio of ginseng to
be
effective should be higher then approximately 1.5 (Figure 10).
In Figure 10, each bar represents ginseng with different diols/triols
ratio. Once a ratio is reached of approximately 1.5 or higher the postprandial
blood glucose is significantly reduced (letter b vs. a on graph). The first
bar
(area= 167) is Chinese ginseng root with ratio 0.8. The second bar (area=
196) is American ginseng extract (75%water: alcohol 25%) with ratio 1.2 . The
third bar (area= 159) is American root ginseng with ratio 1.37.Fourth bar
(area= 159) is American root ginseng with ratio 1.37. The fifth bar represents
(area= 129) is Chinese ginseng extract (59%water: 41 %alcohol) and ratio of
1.91 and this reduced blood glucose significantly. The sixth bar represents
(area= 142) American root ginseng with ratio 1.51 and this also reduced blood
glucose significantly. The seventh bar represents (area= 109) is American
ginseng root with ratio 2.44 and this reduce blood glucose significantly.
A further pilot study in healthy volunteers was conducted to explore the
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effect of the whole root ginseng versus an alcohol:water=55:45 extract,
both with dials to trial ratio > 1.5 with or without the addition of KJM mix.
Results showed that both ginseng types and the KJM lowered the incremental
glucose area and combining the ginseng with the KJM mix had an additional
effect (results not shown).
Example 15 - The Effect of American Ginseng on Insulin And
Nitric Oxide
Other metabolic benefits such as increase in insulin secretion and nitric
oxide generation (Figure 11) are demonstrated with ginseng with ratio of
above 1.5. In this study ginseng had ratios of diols/triols of 2.44. Ginseng
was
administrated to 8 (gender: 3m: 5f, age: 34 3years, BMI: 24.6 0.8kg/m2)
healthy individuals. Data presenting reduction of postprandial blood glucose,
and increase in plasma insulin and nitric oxide generation, relative to
control.
Data are presented in Figure 11.
Comparison of incremental change and area under the curve (AUC) in
plasma glucose, (Figure 11A, 11B), insulin (Figure 11C and 11D) and nitric
oxide (NOx) (Figure 11E and 11F) following ginseng (diols/trials ratio=2.44))
taken 40 min before (r) or together (0) with a 75g oral glucose tolerance test
(75g-OGTT) or a 75g-OGTT done alone previously (g) in 8 non-diabetic
subjects. Glucose and insulin were measured by glucose oxidase method (67)
and double antibody radioimmunoassay method respectively (68). Plasma
NOx was measured as total nitrite (N02) + nitrate (N03) concentrations by
the chemiluminescence method (69,70) using a Sievers 280 NO Analyzer
(Boulder Colorado, USA). Points or bars with different letters are
significantly
different (repeated measures ANOVA adjusted for multiple pairwise
comparisons with the Newman Keuls procedure, P<0.05). Data are
mean SEM.
Example 16 - Effect of ginseng on postprandial glycemia
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The present inventor conducted four studies to determine the
effect of American ginseng in humans were conducted (Table 8).
In the first of these studies, the glycemic responses in 10 normal and 9
type 2 diabetic subjects was studied after the administration of 3g American
ginseng or placebo given 40 minutes before (-40 minutes) or together with a
25g oral glucose challenge. It was observed that selected blood glucose
concentrations and the area under the curve were reduced significantly when
ginseng was administered either before or together with the challenge
compared to placebo in the diabetic subjects and only when given before in
the normal subjects (Vuksan, Arch.lntern. Med 2000; 160:1009-13.
Similar reductions in postprandial glycemia both in nondiabetic and diabetic
subjects were observed, in three subsequent acute dosing and timing
response studies that followed (Table 8). The first of these two studies
showed that 3, 6, or 9g of American ginseng compared to placebo
significantly reduced the postprandial glycemic response to a 25g oral glucose
challenge when administered 0 (together with), 40, 80, or 120 minutes before
a glucose challenge in 10 diabetic subjects (Vuksan, Doabetes Care 2000;
23:1221-6) and when administered 40, 80, or 120 minutes before the
challenge in 10 nondiabetic subjects (Vuksan, J Am Coll Nutr 2000: 18:738-
744). There were no differences observed in either study between the doses
or the times of administration in their glycemic lowering effect. These data
suggested that ginseng is equally effective at doses above 3g and when
administered at any time together or before the challenge in diabetic
subjects,
but only when administered 40 minutes or more before the challenge in
nondiabetic subjects. The third study showed that 1, 2, or 3g of American
ginseng compared to placebo reduced significantly the postprandial glycemic
response to a 25g oral glucose challenge when administered 40 minutes but
not 20, 10, or 0 minutes before a glucose challenge in 12 nondiabetic subjects
(Vuksan Am. J Clin Nutr, ). Again there were no differences detected
between the doses studied. The suggestion was that ginseng is equally
effective at doses above 1g, but needs to be administered a minimum of 40
CA 02410556 2007-10-17
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min before the challenge in nondiabetic subjects.
Having illustrated and described the principles of the invention in a
preferred embodiment, it should be appreciated to those skilled in the art
that
the invention can be modified in arrangement and detail without departure
from such principles. We claim all modifications coming within the scope of
the following claims.
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TABLE 1
Baseline Characteristics of the Study Subjects According to Sex*
MEN WOMEN
CHARACTERISTICS (N = 5) (N = 6)
Age - yr 62 8 59 7
Body weight - % desirablet 133 33 143 22
Android obesity - prevalence $ 5 4
Baseline values:
Serum total cholesterol- mmol/L 6.2 0.4 5.9 0.5
Glycosylated hemoglobin - % 7.4 2.1 8.3 3
Systolic/Diastolic pressure - mm Hg: 139/78 136/82
Known duration of:
Diabetes - yr (self-reported) 11.5 9 18.1 6
Hypertension - yr 7.1 3 6.0 2
Hyperlipidemia - yr 6.3 3 5.6 2
Drug / insulin treatment - prevalence:
Insulin 1 3
Sulfonylurea and/or Metformine 5 6
Diuretics 2 4
Other hypothensive 4 3
Lipid lowering medications 5 6
* Except for drug treatment, blood pressure, and android obesity values are
expressed as
mean SD. To convert values for cholesterol to mg/dI multiply by 38.67.
t Values were assessed from Metropolitan Life Insurance tables, 1983.
t Android obesity is indicated by a wait-to-hip ratio grater than 0.9 for men,
and 0.8 for
women
Bile acid sequestrant, nicotinic acid and/or coenzyme A reductase inhibitor
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TABLE 5
Average Intake of Energy and Nutrients Before and During Study Periods in
Eleven Subjects
Parameters Baseline KJM WB
Total energy (kcaUd) 2 070 700 2 579 628 2 355 420
Total fat (% of energy) 30.5 4.3 29.3 3.2 28.7 2.4
Saturated fat (% of energy) 7.2 4.7 6.7 0.8 6.4 0.7
Monounsaturated fat (% of energy) 10.3 5.1 12.7 2.1 12.2 2.6
Polyunsaturated fat (% of energy) 13.0 5.7 9.9 1.8 10.1 0.9
Cholesterol (mg/d) 328 102 219 48 236 77
Total protein (% of energy) 14.6 8.2 16.2 2.7 15.6 3.2
Available carbohydrate (% of energy) 54.9 21 54.5 9.4 55.7 7.3
Sugars (% of energy) 13.3 3.6 11.2 0.9 9.2 1.4
Total fiber (g/d) 24.2 11 34.7 8.4 33.4 9.6
Water soluble (g/d) 6.9 3.2 23.4 1.7 9.9 3.2*
Water insoluble (g/d) 17.3 7.3 11.2 3.8 23.1 2.6*
Sodium (mg) 5 810 2 384 3 162 648 3 380 647
Potassium (mg) 3 882 713 4 530 611 4 840 872
Calcium (mg) 1 366 193 1.260 238 1 487 446
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Table 8
Study Sample Treatments OGTT AUC P value
Reductions
Study 1 10 NGT 3g AG vs placebo @ Omin - P=NS
(Age: 34 7years, 25g
BMI: 25.6 3kg/m2) 3g AG vs placebo @ -40min 18% P<0.05
9 T2DM 3g AG vs placebo @ 0min 19% P<0.05
(Age: 62 7years, 25g
BMI: 29 5kg/m2, 3g AG vs placebo @ -40min 22% P<0.05
HbAi0: 7.6 0.5%)
Study'2 10 NGT Dosing: 3, 6, or 9g AG vs placebo 26.6, 29.3, P<0.05
(Age: 41 13years, 25g 38.5% for 3,6,
BMI: 24.8 3.5kg/m2) Timing: -40min vs -120 or -80min and 9g P=NS
Study 3 10 T2DM Dosing: 3, 6, or 9g AG vs placebo 19.7, 15.3, 15.9 P<0.05
(Age: 63 2years; 25g % for 3,6, and
BMI: 27.7 1.5kg/m2; Timing: -120, -80, -40 or 0min 9g P=NS
HbAlc: 7.3 0.3%)
Study 4 12NGT Dosing: 1, 2, or 3g AG vs placebo 14.4, 10.6, 9.1 P<0.05
(Age: 42 7 years, 25g % for 1,2, and
BMI: 24.1 1.1 kg/m2) Timing: -40min vs -20, -10 or Omin 3g P<0.05
14.1, 15.0, 9.2
% for -40min
AG, NGT , OGTT, T2DM, NS denote American ginseng, normal glucose tolerance,
oral glucose tolerance
test, type 2 diabetes mellitus, and nonsignificant respectively. P-values are
for repeated measures analysis
of variance (ANOVA) comparisons between absolute values. Values are mean:~SD.
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DETAILED LEGENDS OF THE TABLES
Table 1
* Except for drug treatment, blood pressure, and android obesity values are
expressed as mean SD.
t Values were assessed using Metropolitan Life Insurance tables, 1983.
t Android obesity is indicated by a wait-to-hip ratio grater than or equal to
0.9
for men, and 0.8 for women
Bile acid sequestrants and/or HMG-coenzyme A reductase inhibitors.
Table 2
* Values are mean SD. Konjac-mannan and wheat bran diets are based on
actual intake.
t Based on the mean of four 3-day food records.
t Differences between Konjac-mannan and wheat bran study periods were
calculated by students t-test for paired data.
Table 3
*Values are calculated by difference: 100 - (moisture + protein + fat + total
dietary fiber + ash). Added sucrose was between 37-40% of total available
carbohydrate.
t Average values for dietary fiber in wheat bran and flour analyzed by method
of Prosky et al., 1985.
t Value represents 69% glucomannan polymer derived from KJM flour.
Table 4
*Except for body weight (mean SD), all values are expressed as mean SEM.
t Between treatment differences assessed by ANCOVA (PROC GLM)
t Comparisonwise alpha (a) level was adjusted for multiple endpoint
comparisons with the Bonferroni-Hochberg procedure for primary and
secondary endpoints separately.
Significant after adjustment of alpha level by the Bonferroni-Hochberg
procedure. Null-hypotheses were rejected only if the p-values were less than
their
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corresponding a-value. P-values for during-treatment changes were
assessed by paired t-test
II LDL values are for nine subjects, since two subjects had triglycerides
above
4.5 mmol/L preventing calculation by Friedewald equation.
Table 5: Data are mean SD. KJM+ and WB-control diets are based on
actual intake. Baseline values are based on the mean of four 3-day food
records. *P<0.001 for differences between KJM+ and WB-control treatments
(student's t-test for paired data)
Table 6: Data are expressed as mean SEM, except for body weight which is
mean SD. Within-treatment differences (week-0 versus week-3) were
assessed by paired Student's t-test and between-treatment differences by
ANCOVA (GLM procedure). *Significant after adjustment of alpha level by the
Bonferroni-Hochberg procedure. Null-hypotheses were rejected only if the p-
values were less than their corresponding a-value.
Table 7: Shows the Test Breakfast used in Example 10.
Table 8 Is a comparison of 4 studies. See Example 16.
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FULL CITATIONS FOR REFERENCES REFERRED TO IN THE
SPECIFICATION
Alison K, Ryttig KR, Hylander B, Rossner S: A dietary fibre supplement in the
treatment of mild hypertension. A randomized, double-blind, placebo
controlled trial. J Hypertens 10:195-199, 1992
American Diabetes Association (ADA): Nutrition Recommendations and
principles for people with diabetes mellitus. Diabetes care 22:S42-S43, 1999
Anderson JW, Tietyen-Clark J: Dietary fiber: hyperlipidemia, hypertension,
and coronary heart disease. Am J Gastroenterol 81:907-919, 1986
Aro A, Uusitupa M, Voutilainen E, Hersio K, Korhonen T, Siitonen 0:
Improved diabetic control and hypocholesterolaemic effect induced by long-
term dietary supplementation with guar gum in type 2 (insulin-independent)
diabetes. Diabetologia 21:29-33, 1981
Arvill A, Bodin L: Effect of short-term ingestion of Konjac glucomannan on
serum cholesterol in healthy men. Am J Clin Nutr 61:585-589, 1995
Brown L, Rosner B, Willett WW, Sacks FM: Cholesterol-lowering effects of
dietary fiber: a meta-analysis. Am J Clin Nutr 69:30-42, 1999
Burt VL, Cutler JA, Higgins M, Horan MJ, LaBarthe D, Whelton P, Brown C,
Rocella EJ: Trends in the prevalence, awarness, tretment and control of
hypertension in the adult US population: data from the Health Examination
Surveys, 1960-1991. Hypertension 26:60-69, 1995
DCCT Research Group: The effect of intensive treatment of diabetes on the
development and progression of long-term complications in insulin-dependent
diabetes mellitus. The diabetes control and complications trial. New Engl J
Med 329:977-986, 1993
Doi K, Matsuura M, Kawara A, Baba S: Treatment of diabetes with
glucomannan Konjac mannan. Lancet 1:987-988, 1979
Eastwood MA, Morris ER. Physical properties of dietary fiber that influence
physiological function: a model for polymers along gastrointestinal tract. Am
J
Clin Nutr 55:436-442, 1992
Ebihara K, Masuhara R Kiriyama S: Major determinants Plasma glucose-
flattening activity of a water-soluble dietary fiber: effects of konjac mannan
on
gastric emptying and intraluminal glucose diffusion. Nutr Reports Intl 23:1145-
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