Note: Descriptions are shown in the official language in which they were submitted.
CA 02855291 2014-06-30
METHODS FOR RADIATION PROTECTION
Background
Radiation may be in the form of X-rays, gamma rays, alpha particles, beta
particles,
neutrons, and charged particles. Exposure to damaging or lethal radiation may
result from a
number of sources including, but not limited to, nuclear accidents, wartime or
terrorist nuclear
attack, therapeutic or diagnostic radiology, improper disposal of nuclear
wastes, and outer
space exploration. The extent of radiation-induced internal injury will depend
on the duration,
dose, and type of radiation exposure. Radiation-induced internal injury may
include, but is not
limited to, cell damage, and cell death; and may affect internal processes in
the body such as
the hematopoietic system (due to the reduction in number of hematopoietic
cells such as
lymphocytes, granulocytes, thrombocytes, and reticulocytes), gastrointestinal
system (due to
damage to epithelial cells lining the intestinal tract), and central nervous
system (e.g., due to
damage to neurons, astrocytes and blood vessels in the brain). What is
provided is a method for
protecting and a method for treating an individual against radiation-induced
internal damage.
Summary
A method is provided for protecting an individual against radiation-induced
internal
damage that comprises administering an effective amount of an oral
composition. The oral
composition comprises a mixture of Gynostemma pentaphyllum, Crataegus
pinnatifida
(hawthorn), and Camellia sinensis (green tea) to an individual so that
radiation-induced
internal damage will be prevented or ameliorated. One possible composition
comprises about
to about 30 percent by weight of Gynostemma pentaphyllurn, about 10 to about
30 percent
by weight of Camellia sinensis (green tea), and about 40 to about 75 percent
by weight of
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Crataegus pinnatifida (hawthorn). The composition could comprise a similar
mixture of an
aqueous extracts and alcohol extracts of those components in that proportion.
The radiation
treated under this method is an ionizing radiation selected from the group
consisting of alpha
radiation, beta radiation, gamma radiation, neutron radiation, X-ray and a
combination
thereof. It is possible for the method to be performed prophylactically where
an effective
amount is administered before exposure to radiation. This method could be
administered at
one or more times of during radiation exposure, and after radiation exposure.
It is also
possible for the oral composition to be administered in multiple doses. This
method could
also be administered to the individual prior to expected exposure to
radiation, during
exposure to radiation, or after exposure to radiation.
Detailed Description
It will be understood that variations in the embodiments can generally be
interchanged
without deviating from the invention.
Definitions: While the following terms are believed to be well understood by
one of
ordinary skill in the art of biotechnology, the following definitions are set
forth to facilitate
explanation of the invention.
The "oral composition comprised of a mixture of Gynostemma pentaphyllum,
Crataegus
pinnatifida, and Camellia sinensis" comprises about 10 to about 30 percent by
weight of
Gynostemma pentaphyllum, about 10 to about 30 percent by weight of Camellia
sinensis (green
tea), and about 40 to about 75 percent by weight of Crataegus pinnatifida
(hawthorn leaves
and/or berries). Although there may be various methods to combine these
ingredients, one
method of making the oral composition is disclosed in U.S. Patent No.
5,910,308. The
composition comprises about 10 to about 30 percent by weight of Gynostemma
pentaphyllum
extract, about 10 to about 30 percent by weight of green tea extract, and
about 40 to about 75
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percent by weight of hawthorn extract. The preferred composition comprises
about 10 to about
30 percent by weight of a mixture of an aqueous extract and an alcohol extract
of Gynostemma
pentaphyllum, about 10 to about 30 percent by weight of a mixture of an
aqueous extract and an
alcohol extract of green tea, and about 40 to about 75 per-cent by weight of a
mixture of an
aqueous extract and an alcohol extract of Crataegus pinnatifida (hawthorn
leaves and/or berries).
Gynostemma pentaphyllum, also known as 5-leaf ginseng or Jiaogulan or southern
ginseng, is
from the cucumber family and is rich in special saponins termed "gypenosides"
some of which
are similar, and some of which are different, to the ginsenosides found in
ginseng, but at a level
several fold higher. The leaves and berries of Crataegus pinnatifida, also
known as hawthorn,
contain saponins, flavonoids (including hyperoside), and anthocyanins
(including
proanthocyanidins). Leaves from the Camellia sinensis plant, particularly when
processed into
green tea, contain polyphenols including catechins such as epigallocatechin-3
gallate (EGCG),
epigallocatechin, and epicatechin-3-gallate. While Gynostemma pentaphyllum,
Crataegus
pinnatifida, and Camellia sinensis have been used individually for health
promoting and
therapeutic purposes, not described is the ability of a composition comprising
a mixture of
Gynostemma pentaphyllum, Crataegus pinnatifida (hawthorn) and Camellia
sinensis (green tea)
to work together synergistically to protect an individual against radiation-
induced internal
damage. Optionally, the oral composition contains one or more carriers to
facilitate one or more
of formulation and administration.
The term "individual" is used herein, for purposes of the specification and
claims, to
mean an animal, preferably a mammal, and more preferably a human.
The term "radiation-induced internal injury" refers to injury or damage caused
by
radiation exposure, wherein the injury or damage is inside the body of an
individual exposed to
radiation; i.e., excluded is sunburn.
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The term "ionizing radiation" is used herein, for purposes of the
specification and claims,
to mean radiation that has sufficient energy to eject one or more orbital
electrons from an atom or
molecule (e.g., a particles, 13 particles, y particles, neutrons, protons, x-
rays).
The terms "treatment" or "treating" are used herein, for purposes of the
specification and
claims, to mean administration of the oral composition in an amount that is
effective in
preventing, reducing or ameliorating radiation-induced injury to an individual
who has been, is
being, or will be exposed to radiation (thus, the oral composition being
"radioprotective"). In the
case that an individual will be exposed to radiation, the treatment is
prophylactic (i.e., prior to
radiation exposure, the oral composition is ingested by the individual using a
prophylactically
effective amount). In the case that that an individual is being or has been
exposed to radiation,
the treatment is therapeutic; i.e., after radiation exposure, the oral
composition is ingested by the
individual using a therapeutically effective amount.
Radiation-induced injury may include internal injury or internal damage to the
human
body caused by exposure to radiation. Such internal injury may include, but is
not limited to,
bone marrow cell damage, intestinal damage, damage to the central nervous
system, DNA
mutations causing cell injury and/or death, and development of cancer. The
methods are
particularly useful for treating an individual (including a single individual
or more than one
individual) engaging in activities involving a high risk of radiation
exposure. Also, the methods
can be used to treat individuals exposed to radiation so as to prevent or
ameliorate radiation-
induced injury. Exposure to radiation may be a result of radiation emanating
from radioactive
materials released by terrorists or as a result of a nuclear accident, from
diagnostic machines such
as an X-ray machine, a C-T scanner, or a synchrotron, from therapeutic
radiation (e.g., intended
to kill cancer cells, wherein the radiation-induced injury to be prevented or
ameliorated is to the
nonmalignant tissue, surrounding or adjacent to the tumor intended to be
killed by radiation).
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For use in a method, the oral composition is formulated in any acceptable oral
dosage
form, including but not limited to, a tablet, pill, caplet, capsule, lozenge,
powder, solution,
suspension, and the like for ingestion. Oral compositions comprising a
tablets, pills, caplets,
capsules, or lozenges generally include carrier. Such a carrier may include
one or more of a
binder (e.g., such as, but not limited to, polyol, microcrystalline cellulose,
gelatin, gum such as
arabic or tragacanth, etc.), an excipient (e.g., such as, but not limited to,
starch, lactose, calcium
carbonate, sodium citrate, calcium phosphate, etc.), a disintegrant (e.g.,
such as, but not limited
to, alginic acid, corn starch, silicates, etc.), a lubricating agent (e.g.,
such as, but not limited to,
talc, sodium lauryl sulfate, magnesium stearate or other stearates, etc.), a
coating (e.g., such as,
but not limited to, lecithin, etc.), and a flavoring agent (e.g., a sweetener
such as a carbohydrate,
sugar alcohol, saccharin, aspartame, stevia, and the like; or fruit or mint
flavoring). Where the
oral composition is liquid in nature or containing a liquid, the carrier may
be a solvent or
dispersion medium including but not limited to one or more of an inert
diluent, water, an alcohol,
an edible oil, and a syrup.
In the method for protecting an individual against radiation-induced internal
damage, the
oral composition comprising a mixture of Gynostemma pentaphyllum, Crataegus
pinnatifida,
and Camellia sinensis is administered to an individual that may be potentially
exposed to
radiation (e.g., as a result of engaging in an activity that has a high risk
of radiation exposure), an
individual that is in the process of being exposed to radiation, or an
individual who has already
been exposed to radiation. The effective amount of the oral composition to
administer to the
individual in the method will vary depending on several factors including, but
not limited to, the
age, general health, and body weight of the individual at the time of
administration; and the
severity of the radiation exposure, expected radiation exposure, or apparent
radiation-induce
injury. Additionally, an effective dose may be determined by standard
pharmaceutical procedures
CA 02855291 2014-06-30
such as using cell cultures, or experimental in vivo models, and by the
medical profession (e.g.,
doctor, nurse, pharmacist, pharmacologist, etc.) taking into consideration
factors relating to the
individual in need of treatment.
The radioprotective oral composition may be administered once per day, or
multiple
times (e.g., 2 to 5 times) per day, as needed, more preferably 2 to 3 times
per day, as needed, or 3
times per day, to an individual in need of such treatment. Preferably, during
each administration
of a dose, 1 to 3 tablets, caplets, capsules, pills or other form of the oral
composition are ingested
by the individual in need of treatment. The active ingredients in the oral
composition are
components in the 5 mixture of Gynostemma pentaphyllum, Crataegus pinnatifida,
and Camellia
sinensis. A dosage amount of the oral composition for each administration may
contain from
about 0.1 gram to about 1 gram of the active ingredients, and more preferably
from about 0.2
grams to about 0.75 grams of the active ingredients.
The examples presented herein are intended to be illustrative in nature, and
in no way
intended to limit the scope of the claimed methods and discussed in detail
above. In these
examples, four methods were used to illustrate a method; i.e., use of an oral
composition
comprising a mixture of Gynosternma pentaphyllum, Crataegus pinnatifida, and
Camellia
sinensis can work synergistically to prevent or ameliorate radiation-induced
injury or damage.
Bone marrow failure is the major cause of radiation lethality in mammals. An
accepted and
standard indirect method to determine the consequences of damage to marrow
resulting from
radiation exposure includes measurement of peripheral blood cells, wherein
reductions in
peripheral blood cell counts reflect radiation damage.
The second method utilized in this assessment is the micronucleus assay.
Ionizing
radiation is a strong clastogenic agent, and thus a potent inducer of
micronuclei (chromosomal
aberrations, typically the result of unrepaired or misrepaired double-strand
DNA breaks). The
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micronucleus assay quantifies radiation-induced chromosome damage expressed as
post-mitotic
micronuclei. Many studies have shown that the number of radiation-induced
micronuclei is
strongly correlated with dose and type of radiation. Thus, the micronucleus
assay has become a
validated and standard technique for evaluating in vivo radiation exposure of
individuals, as well
as for determining the radioprotective effect of a test compound.
The third method used to illustrate a method of the invention is the comet
assay or single
cell gel-electrophoresis assay. The assay is based on the embedding of cells
in agarose, followed
by cell lysis, and subsequent electrophoresis. The electric current pulls the
charged DNA from
the nucleus, where broken DNA fragments migrate further from the nucleus than
intact DNA.
The resultant pattern, named for its resemblance to comets, is used to measure
and determine the
extent of DNA damage. For example, it is known that comet parameters such as
tail length, tail
moment, and percent of DNA in tail were increased in the blood leukocytes
exposed to high y -
radiation. When a radioprotective agent is administered before the radiation
exposure, the
increase in the comet parameters as a result of radiation was prevented,
indicating a protection of
cellular DNA. Thus, the comet assay is a validated and standardized technique
for determining
the radioprotective effect of a test compound.
The fourth method is measurement of total antioxidant capacity (T-AOC). It is
known
that ionizing radiation exposure induces free radicals which damage cells.
Scavenging of free
radicals produced from radiation exposure can protect cells from radiation-
induced injury or
death. This assay can be used to ascertain whether or not the mechanism of
action of a
radioprotectant involves free radical scavenging.
EXAMPLE 1
In this Example, illustrated are methods of radioprotection using an oral
composition
comprising a mixture of Gynostemma pentaphyllum, Crataegus pinnatifida, and
Camellia
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sinensis to treat radiation-induced injury or damage in standard models for
determining a
radioprotective effect. Male BALB/c mice, weighting from 22g to 28g, were
divided into a two
groups, A "treatment" group was pre-treated by administering an oral
composition comprising a
mixture of Gynostemma pentaphyllum, Crataegus pinnatifida, and Camellia
sinensis. The oral
composition, comprising a mixture of Gynostemma pentaphyllum, Crataegus
pinnatifida, and
Camellia sinensis as previously described herein in more detail, that was used
in these
experiments is commercially available (ONCOLYN ). Mice in the treatment group
were treated
with the oral composition at dose of 500mg/kg for 7 days by oral gavage. The
"control" group of
mice was administered distilled water as the same volume and timing as was
administered the
oral composition to the treatment group.
Whole body irradiation of each of the treatment group and control group was
performed
with an X-ray source. Mice were placed in ventilated caged and irradiated in
groups of five
mice simultaneously. The source to skin distance was 100 cm with a dose rate
of 300 cGy/min
at room temperature. The mice were irradiated with a total dose of 1.5 Gy or
3.0 Gy. At 24
hours after irradiation, mice were anesthetized. Blood samples were taken from
abdominal
aorta. 0.5m1 blood was put into heparin for anticoagulation. Blood cell
classification and
counting were performed using an automated cell analyzer. As shown in Table,
1, a method of
the invention for protecting an individual against radiation-induced internal
damage comprises
administering the oral composition comprising a mixture of Gynostemma
pentaphyllum,
Crataegus pinnatifida, and Camellia sinensis to an individual that is
subsequently exposed to
radiation. As shown in Table 1, the treatment group (receiving the oral
composition comprising
a mixture of Gynostemma pentaphyllum, Crataegus pinnatifida, and Camellia
sinensis) prior to
radiation exposure showed a radioprotective effect as evident by a
significantly increased
number of peripheral blood cells as compared to that of the control group
which received the
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corresponding dose of radiation but did not receive treatment with the oral
composition. In
Table 1, the total number of cells counted is expressed in 109/L, wherein
"WBC" is the white
blood cell count; "GRAN" is the granulocyte count; "LYM" is the lymphocyte
count; "MID" is
the count of less frequently occurring and rare cells correlating to
monocytes, eosinophils,
basophils, blasts and other precursor white cells.
Table 1- Radiation-induced internal damage of bone marrow, as measured by
peripheral
blood cell counts
Groups WBC GRAN LYM MID , platelets
No radiation 10.8911.19 1.5410.38 5.1311.24
1.740.47 578.60171.50
3.0Gy Control 4.4710.87* 0.6710.19*
2.8010.57* 0.8410.19* 313.25 67.67*
3.0Gy Treatment 8.53 0.92* 1.8110.32 5.0811.08* 1.78 0.51
415.14 55.25*
1.5Gy Control 7.32 1.36* 1.03 0.24* 4.75 0.79
1.08 0.23* 494.16 82.83
1.5Gy Treatment 10.40 2.01* 1.65 0.31# 5.36 1.06 1.8710.64*
503.57184.33
Notes: *- control group compared with non-radiated animals p<0.05;
#- treatment group compared with control group and at the corresponding
radiation dose p<0.05.
A standard comet assay was performed from the blood samples collected. The
entire
assay was conducted under low indirect incandescent light (60 Watts) to
minimize light-induced
damage to lymphocyte DNA. Two fully-frosted microscopic slides per sample were
prepared.
Each slide was covered with 100 pi of 1% normal melting agarose at about 42 C
in phosphate
buffered saline (PBS), and the gel was immediately covered with a cover slip
and kept for 20
minutes in a refrigerator to solidify. After gently removing the cover slip,
100 1 mixture of 30 pl
blood sample in PBS and 90 Al of 1% low melting agarose was rapidly add onto
the agarose
layer, and then spread using a cover slip. The cover slips were then removed,
and the slides were
submersed in freshly prepared cold lysis solution (2.5M NaCI, 100 mM Na2EDTA,
10mM Tris
base, with freshly added 1% Triton.' X-100 and 10% DMSO, pH=10) at 4 C for at
least 1 hour.
Slides were then placed in a horizontal electrophoresis tank with freshly
prepared alkaline
electrophoresis solution (1 mM Na2EDTA, 300 mM NaOH, pH>13) for 30 minutes at
4 C to
allow the unwinding of the DNA, and expression of alkali-labile site, before
being
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electrophoresed under neutral conditions at 12 volts and 300 milliamps for 30
minutes at room
temperature using an electrophoresis compact power supply. When put in an
electrical field, the
intact DNA was such a large molecule that it hardly moved. DNA breaks,
however, lead to
smaller pieces of DNA which migrated away from the intact DNA. After
electrophoresis, the
slides were equilibrated in neutral solution (300 mM sodium acetate, 100 mM
Tris, pH 9) for 5
minutes. The slides were stained with ethidium bromide (20 mg1-1,
351.d/slide). Quantification of
DNA damage was assessed in over 100 cells in the center of each gel, by using
a microscope
(equipped with a 100-W mercury lamp and WG filter block), and taking pictures
at 400x
magnification. The cells containing damaged DNA had the appearance of a comet
with a bright
head (undamaged) and tail. Measurements of Comet parameters included presence
of comets, tail
length. About 1000 cells from each slide were examined by comet image analysis
software was
used to measure comet extent (tail length) and percentage of comet cells.
Comet extent is a
measure of total comet length from the beginning of the head to the last
visible pixel in the tail.
The percentage of comet cells is calculated by dividing the comet cell number
counted in 100
lymphocytes per slide by 100 lymphocytes.
A standard micronucleus assay was performed. Peripheral blood cells were
smeared, and
then immediately stained with a DNA specific stain (e.g., acridine orange).
The proportion of
immature among total (immature + mature) erythrocytes is determined for each
animal by
counting a total of at least 1000 erythrocytes for peripheral blood. At least
2000 immature
erythrocytes per animal are scored for the incidence of micronucleated
immature erythrocytes.
As shown in Table 2, a method for protecting an individual against radiation-
induced
internal damage comprises administering the oral composition comprising a
mixture of
Gynosternma pentaphyllurn, Crataegus pinnatifida, and Camellia sinensis to an
individual.
As shown in Table 2, as compared to the control group with the corresponding
radiation
CA 02855291 2014-06-30
exposure, treatment with the oral composition resulted in a significant
reduction in the
treatment group of the: (a) number of micronucleated cells; (b) percentage of
comet cells; and
(c) tail length of comet cells (i.e., demonstrated is a radioprotective
effective).
Table 2- Radiation-induced internal damage, as measured by micronucleus assay
and by
the comet assay.
Groups Micronucleated % comet Tail length of
cells comet cells (pm)
cells (Too)
no radiation 2.83 0.75 4.2 0.91 11.45 2.56
3.0Gy X-ray control 19.5 5.68* 42,67 7.88* 42.67 6.58
3.0Gy X-ray treatment 11.14 4.89# 25.47 5.464 27.19 4.73
1.50y X-ray control 14.67 4.12* 30.13 7.94* 31.56 5.33
1.5Gy X-ray treatment 9.50 3.03# 17.68 6.394 22.20 4.11
Notes: * control group compared with non-irradiated animals, p<0.05;
# treatment group compared with the control group of the corresponding
radiation dose, p<0.05
A commercially available assay for measuring total antioxidant capacity was
used.
Briefly, serum from mice of a control group or from mice of a treatment group
is mixed with a
reagent containing pholasin, and incubated for 30 minutes at 37 C.
Antioxidants can reduce Fe3+
to Fel', and Fe2+ binds with pholasin which produces a visible chelating agent
measurable at an
absorption of 520 nm. An increase in 0.01 of the absorption value per minute
per milliliter serum
was determined as a unit of total antioxidant capacity. Total antioxidant
capacity unit= [(Optical
Density of the determined tube - Optical Density of the assay control tube) x
N x n] 0,01;
wherein "N" is the diluted fold in the reaction system (total volume of
reaction/serum sample
volume), and "n" is the fold dilution of sample. As shown in Table 3, a method
for protecting an
individual against radiation-induced internal damage comprises administering
the oral
composition comprising a mixture of Gynostemma pentaphyllum, Crataegus
pinnatifida, and
Camellia sinensis to an individual. As shown in Table 3, as compared to the
control group with
the corresponding radiation exposure, treatment with the oral composition at
the higher doses of
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radiation (e.g., 3.0Gy) resulted in a significant increase in the total
antioxidant capacity in the
treatment group. This suggests that at radiation doses approaching 3.0Gy or
greater, the free
radical scavenging (antioxidant activity) of the oral composition is
contributing to the method of
protecting against radiation-induced internal damage. The results also suggest
that after certain
doses of radiation exposure (e.g., 1.5Gy), the mixture in the oral composition
works
synergistically by mechanisms in lieu of or in addition to free radical
scavenging in protecting an
individual from radiation-induced internal injury or damage. Such other
mechanism(s) could
include, but is not limited to: protecting DNA from radiation-induced damage;
enhancing repair
of DNA damage induced by radiation; inhibiting apoptotic pathway(s) induced by
radiation or
radiation-induced damage; inhibiting radiation-induced activation of cell
signaling pathways;
activating of radiation-induced inhibition of cell signaling pathways; or a
combination thereof.
Table 3 Measurement in serum of total antioxidant capacity (T-AOC)
Groups T-A0C(U/m1)
No radiation 11.99 3.66
1.50y X-ray control 9.18 3.04
1.5Gy X-ray treatment 9.46 1.89
3.0Gy X-ray control 7.03 1.82*
3.0Gy X-ray treatment 12.26 2.42#
Notes: * compared with non-irradiated animals (assay control) p41.05;
# Treatment group compared with the corresponding X-ray Control group p41.05
EXAMPLE 2
In this Example, illustrated are methods of radioprotection using an oral
composition
comprising a mixture of Gynostemma pentaphyllum, Crataegus pinnatifida, and
Camellia
sinensis to treat radiation-induced internal injury or damage in an individual
for determining a
radioprotective effect. Eight human volunteers, aging from 25 years to 30years
in age, gave
blood samples, and then took 1 gram of an oral composition, comprising a
mixture of
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Gynostemma pentaphyllum, Crataegus pinnatifida, and Camellia sinensis as
previously
described herein in more detail. Following the administration of the oral
composition, the human
volunteers gave blood samples at 1 hour and also at 3 hours post treatment
with the oral
composition. All blood samples were then subjected to radiation exposure (1,5
Gy y -radiation).
A portion of each blood sample was used for a comet assay test, and the
remainder of each blood
sample was used for the micronucleus test (using the assay protocols
essentially described in
Example 1 herein).
For the comet assay, the lymphocytes were separated from polymorphonuclear
leucocytes
and erythrocytes contained in the heparinized whole blood sample, and the
lymphocytes were
then washed twice and suspended in 500 pi ice-cold phosphate-buffered saline
(PBS). The comet
assay was performed on the lymphocytes isolated from each blood sample. As
shown in Table 4,
a method for protecting an individual against radiation-induced internal
damage comprises
administering the oral composition comprising a mixture of Gynostemma
pentaphyllum,
Crataegus pinnatifida, and Camellia sinensis to an individual. As shown in
Table 4 and Table 5,
as compared to the control group with the corresponding radiation exposure,
treatment with the
oral composition resulted in a significant reduction in the treatment group of
the: (a) percentage
of comet cells; and (b) tail length of comet cells (i.e., demonstrated is a
radioprotective
effective). In Table 4 and Table 5, "Individual" represents the different
individuals who gave
blood samples for treatment; "baseline" represents comet assay results on
blood samples which
were not exposed to radiation nor treatment; Hy ray control" represents comet
assay results from
blood samples exposed to radiation but not treated; and "7 ray post treatment"
represents comet
assay results from blood samples from individuals treated with the oral
composition, which
blood samples were subsequently exposed to radiation after the designated
number of hours.
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Table 4- Radiation-induced internal damage, as measured by comet cell number
(%)
(%)ual Individ y ray post y ray post
baseline y ray control treatment 1 treatment 3
hours hours
1 5.2 1.1 62.57115.44 40.12 8.57* 22.6616.1244
2 4.8 10.92 58.79111.09 41.23 9.66* 23.5815.72*#
3 5.711.1 59.87110.38 38.6417.38* 19.6718.97*#
4 5.711.0 61.72115.44 42.71 9.37 25.4116.77*#
6.211.3 68.99117.78 46.3519.16* 27.3 2 5.64*
6 5.011.3 54.12110.23 34.7216.97* 18.67 5.12*#
7 5.2 1.4 56.3818.67 39.6617.82* 19.6817.53*#
8 5.811.5 61.46114.55 40.1919.68* 20.1813.22*#
Notes: *- post treatment group compared with y ray control group, p<0.05;
# post treatment 3 hour group compared with y ray control group and with y ray
post treatment- 1 hour group, p<0.05
Table 5- Radiation-induced internal damage, as measured by tail length of
comet cells
(lun
y ray post y ray post
Individual baseline y ray control
treatment 1 treatment 3
hour hours
1 10.45 2.56 72.35115.12 49.6716.58*
32.1914.73*#
2 11.2312.35 68.95114.32 47.26 7.85*
29.6715.06*#
3 10.7213.01 71.26116.38 46.28 6.57
25.64 6.31*4
4 11.3811.47 70.23114.89 48.6718.77
27.0915.68*#
5 12.5912.84 69.26110.22 - 47.66 8.31*
22.59 6.80*#
6 10.6711.29 70.43111.76 42.39 8.05*
21.3814.52*#
7 10.98 2.03 67.91116.34 46.1219.60*
24.3115.30*#
8 11.6512.31 68.55110.33 41.2816.38*
22.37 6.01*#
Notes: *- post treatment group compared with y ray control group, p<0.05;
# post treatment 3 hour group compared with y ray control group and with y ray
post treatment- 1 hour group, p<0.05
This invention has been described with reference to several preferred
embodiments.
Many modifications and alterations will occur to others upon reading and
understanding the
preceding specification. The scope of the claims should not be limited by the
preferred
embodiments set forth in the examples, but should be given the broadest
interpretation
consistent with the description as a whole.
14
