DV Biologics, a Leading Biological company, Reports
Earth Science Tech (ETST) High Grade Hemp CBD (Cannabidiol) Oil
Deters the Formation of Reactive Oxygen Species (ROS) on Human
Brain Cells: May serve as a Neuroprotectant
BOCA RATON, FL -- August 18, 2017 -- InvestorsHub NewsWire
-- Earth Science Tech, Inc. ("ETST"
or "the Company"), a unique biotechnology company focused on
nutraceuticals, bioceuticals and dietary supplements, announced
today that DV Biologics, a leading biological company, has
confirmed ETST High Grade Hemp CBD (Cannabidiol) Oil deterred the
formation of reactive oxygen species and cell death in human brain
cells in vitro. The goal of these studies is to test if
ETST High Grade Hemp CBD (Cannabidiol) Oil, as a dietary
supplement, serves as a neuroprotectant which may help with
neurodegenerative disorders such as Alzheimer's and Parkinson's.
The scientific study which concludes the in vitro
experiments done independently on human brain cells by DV
Biologics, a company that provides biological tools and research
for the advancement of medicine, illustrates that using ETST High
Grade Hemp CBD (Cannabidiol) Oil can rescue human brain cells from
the detrimental effects of reactive oxygen species and free radical
formation which cause damage to brain cells (see study below).
Recently, independent studies demonstrated that ETST High Grade
Hemp CBD (Cannabidiol) Oil can reduce lipid peroxidation (oxidative
stress) on human brain cells following the addition of hydrogen
peroxide. This new data supports and extends the previous data that
ETST High Grade Hemp CBD (Cannabidiol) Oil may help serve as a
neuroprotectant. This study looks at another pathway of free
radical formation, oxidative stress and cell death that occurs in
the brain during neurodegenerative disorders such as
Alzheimers and Parkinsons. This data suggest that ETST High Grade
Hemp CBD (Cannabidiol) Oil may have neuroprotective properties.
"Here at ETST we will continue focusing on providing the best
All-Natural High Grade Hemp CBD (Cannabidiol) Oil as a dietary
supplement. We believe the only way to do this is by studying the
effects of ETST High Grade Hemp CBD Oils in various
experiments. Science is what will show the world that ETST provides
high quality products as well as the best All-Natural High Grade
Hemp CBD (Cannabidiol) Oil designed to give consumers maximum
overall wellness benefits. We have relatively exciting studies
which we will be starting soon in our continual goal of providing
the best products for consumers worldwide and great value to
our investors," stated Matthew Cohen, CEO of ETST.
According to Dr Rafael Gonzalez, VP of Research & Development
at DV Biologics, "The data speaks for itself. This in
vitro study is very encouraging as it demonstrates that ETST
High Grade Hemp CBD (Cannabidiol) Oil may be beneficial for the
brain. The great thing is that the data is supported by 3 different
experiments, all showing the same result that it works. With this
data if I had a neurodegenerative disorder such as Parkinsons, I
would seriously consider taking this as there isnt much available
that can deter the progression of the disorder. We hope that ETST
will continue to give us the opportunity to work with their High
Grade Hemp CBD (Cannabidiol) Oil."
*Complete DV Biologics study shown below*
Does Earth Science Tech High Grade Cannabidiol (CBD)
Rich Hemp Oil Have a Neuroprotective Effect? The DV Biologics
Report and data below is encouraging and clearly demonstrates that
Earth Science Technology (ETST)
High Grade HEMP CBD (Cannabidiol) Oil is protective against lipid
peroxidation in human brain cells in vitro.
The Brain
The brain contains trillions of cells. From neurons with its
long axons (nerve fibers) which can extend greater than 2 feet and
serve to communicate to glial cells which support the brain in all
its complex task. Two-thirds of the brain is composed of lipids.
The vast majority of the different cell types in the brain require
lipids for correct transmission of signals, providing energy and
structural support to our cell membranes. Cells in the body and
particularly in the brain are under constant attack by free
radicals and oxidative stress. There are protective mechanisms that
help deter this normal damage that occurs; however, with aging the
protective mechanism declines substantially and degeneration of the
brain occurs.
Lipid Peroxidation
Lipid peroxidation is the degradation of lipids that occurs as a
result of oxidative damage and is a useful marker for oxidative
stress. Lipids are susceptible to an oxidative attack, typically by
reactive oxygen species, resulting in a well-defined chain reaction
with the production of end products such as malondialdehyde
(MDA).
Reactive oxygen species (ROS)
Reactive oxygen species are molecules that contain oxygen that
are chemically active (i.e. oxygen ions and peroxides). They are
formed as a natural byproduct of metabolism in our bodies. However,
during stress and disease or dysfunction, these byproducts are not
taken removed correctly and damage occurs in our body. The harmful
effect of ROS occurs in cells in the form of DNA damage and
oxidation of amino acids in proteins. ROS come in various forms
such as superoxides and hydroxyl radicals and it is well documented
that ROS play a crucial role in the formation of neurodegenerative
disorders, cancer and aging.
Experiment:
In this experiment, we look at three (3) different
parameters:
- 1. Fluorometric intracellular ROS with a Kit that provides a
sensitive, one-step fluorometric assay to detect intracellular ROS
(especially superoxide and hydroxyl radicals) after induction of
peroxidation with hydrogen peroxide (with and without ETST High
Grade Hemp CBD (Cannabidiol) Oil).
2. Lipid peroxidation is determined by the reaction of MDA with
thiobarbituric acid (TBA) to form a fluorometric complex,
proportional to the amount of MDA present, after induction of
peroxidation with hydrogen peroxide (with and without ETST High
Grade Hemp CBD (Cannabidiol) Oil).
3. Neutral Red Assay to determine viability of cells after
induction of peroxidation with hydrogen peroxide (with and without
ETST High Grade Hemp CBD (Cannabidiol) Oil).
Experimental procedure ROS:
- *Cells were plated in 96 well until it reach 90%. Neural
cells were at a density of 2 X 104 cells/cm2. The cells were grown
in neural media supplemented with 20 ng/ml of epidermal growth
factor (EGF) and basic fibroblast growth factor (bFGF).
*Media was changed every 2-3 days.
*ETST Hemp CBD/Neural-Growth media was prepared by diluting the
ETST Hemp CBD in dimethyl sulfoxide (DMSO). The suspension was
filtered with a 0.22 micron filter. The ETST Hemp CBD suspension
was then diluted in N-GRO media to contain the following
concentrations of Hemp CBD: 5 M, 2.5 M, and 1 M CBD.
*Cells were incubated with ETST Hemp CBD for 48 hours prior to the
addition of hydrogen peroxide.
*After 48 hours, ROS was induced by addition of H2O2 at
concentration of 25 M to certain wells in quadruplicate. H2O2 was
added to the wells of a 96 well plate according to the following
design:
- *Set up the Master Reaction Mix.
*Add 100 mL/well (96 well plate) Master Reaction Mix into the cell
plate.
*Incubate the cells in a 5% CO2, 37 C incubator for one hour.
*Treat cells with 20 mL/well of H2O2 (96 well plate).
*For control wells (untreated cells), PBS was used.
*To induce ROS, incubate the cell plate in a 5% CO2, 37 C incubator
for 16 hours.
*Measure the fluorescence intensity at lex = 490/lem = 525 nm for 1
second/well.
Experimental Procedure Lipid
Peroxidation:
- *Two 24-well plates were coated with Matrigel for 1 hour at 27
Celsius.
*The plates were seeded with human neural cells at a density of 2 X
104 cells/cm2. The cells were grown in neural media supplemented
with 20 ng/ml of epidermal growth factor (EGF) and basic fibroblast
growth factor (bFGF).
*Media was changed every 2-3 days.
*ETST Hemp CBD/Neural-Growth media was prepared by diluting the
ETST Hemp CBD in dimethyl sulfoxide (DMSO). The suspension was
filtered with a 0.22 micron filter. The ETST Hemp CBD suspension
was then diluted in N-GRO media to contain the following
concentrations of Hemp CBD: 10 M, 5 M, 2.5 M, and 1 M CBD.
*After the cells reached 80-90% confluency, base media was replaced
with media containing ETST Hemp CBD or control media; the controls
were media only, media with DMSO 1:100, and media with hydrogen
peroxide (H2O2) (H2O2 was added during induction phase).
*The cells were incubated under experimental conditions for 48
hours in media containing ETST Hemp CBD.
*After 48 hours, lipid peroxidation was induced by addition of H2O2
at concentration of 100 M to certain wells in duplicates. H2O2 was
added according to the following protocol:
Well ID
|
A
|
B
|
C
|
D
|
E
|
F
|
1
|
Media only (control)
|
1 M CBD + H2O2
|
5 M CBD + H2O2
|
2
|
Media +DMSO (control)
|
Media+2.5M CBD only
|
Media+10M CBD only
|
3
|
Media + H2O2
|
2.5 M CBD + H2O2
|
10 M CBD + H2O2
|
4
|
Media+1M CBD only
|
Media+5M CBD only
|
|
|
|
|
|
|
|
|
- *The cells were incubated with H2O2 for 120 minutes at 37
Celsius.
*Media was aspirated and the cells were lysed.
*Total protein was precipitated, and reaction to form MDA-TBA
adduct was performed. The reactions were placed in a 96-well
plate.
*MDA fluorometric detection was done by measuring fluorescence
intensity (excitation 532 nm/ emission 553 nm) of samples and
standard curve.
*Relative lipid peroxidation was measured by subtracting blank
value from all readings.
Experimental Procedure Neutral Red
Assay:
- *Human prenatal neural cells were seeded in a 24 well flask
coated with Matrigel.
*The cells were grown at 37 C, 90 % humidity, and 5.0 % CO2 until
they reach 80% confluency.
*ETST Hemp CBD/Neural-Growth (N-GRO) media was prepared by diluting
the ETST Hemp CBD in dimethyl sulfoxide (DMSO). The suspension was
filtered with a 0.22 micron filter. The ETST Hemp CBD suspension
was then diluted in N-GRO media to contain the following
concentrations of Hemp CBD: 5 M, 2.5 M, and 1 M CBD.
*Cells were incubated with CBD for 48 hours prior to the addition
of H2O2.
*After 48 hours, H2O2 was added at concentration of 25 M to certain
wells in Duplicate. H2O2 was added according to the following
protocol:
|
1
|
2
|
3
|
4
|
5
|
6
|
A
|
Control
|
1uM CBD + H2O2
|
5uM CBD + H2O2
|
|
B
|
DMSO control
|
2.5uM CBD
|
Blank
|
|
C
|
Media+ H2O2
|
2.5uM CBD + H2O2
|
|
|
D
|
1uM CBD
|
5uM CBD
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
- *The cells were incubated with H2O2 for 16 hours at 37 C.
*After incubation, media was aspirated from all wells and 100uL of
NR medium was added to each well.
*Incubate for 1 hour with Neutral red (NR) medium at 37 C.
*NR media was aspirated from all wells and washed each well with
150uL PBS
*100 l of lysis solution (ETOH/acetic acid) solution was added to
all wells, including blanks.
*Plate was shaken rapidly on a microtiter plate shaker for 20
45 min to extract NR
*Lysis solution was transferred to a 96-well plate and absorbance
was determined at 540nm in quadruplicate.
Results
Figure Legend:
Earth Science Tech (ETST) High Grade CBD (Cannabidiol) Oil
rescues human brain cells from reactive oxygen species in vitro.
Human brain cells were cultured and treated with ETST High Grade
CBD (Cannabidiol) Oil diluted to concentrations indicated on the
graph for 2 days (1.0 M CBD = 8 ppm CBD rich hemp Oil; 2.5 M CBD =
20 ppm CBD rich hemp oil; 5.0 M CBD rich hemp oil= 41 ppm Hemp
Oil). The cells were treated with 25 M hydrogen peroxide (H2O2) for
16 hours and incubated with reactive oxygen species (ROS) master
mix reagents for 1 hour to detect intracellular ROS. The
amount of ROS was determined by fluorescence intensity at lex =
490/lem = 525 nm. The experiment is performed in
quadruplicates, with error bars representing standard deviation
error. Control: DMSO only. Positive control: Media + H2O2 +
DMSO.
When brain cells are incubated with H2O2 + DMSO intracellular
ROS accumulates and is much higher than normal physiological states
as seen in the figure. ETST Hemp CBD on its own maintains
intracellular ROS to similar levels as seen in physiological
states. When ETST Hemp CBD is used prophylactically at all
concentrations used (1, 2.5 and 5 M) there is a decrease in the
formation of ROS.
Figure legend:
Earth Science Tech (ETST) High Grade CBD (Cannabidiol) Oil is
protective against lipid peroxidation in human brain cells in
vitro. Human brain cells were incubated with media (control),
hydrogen peroxide (H2O2) to cause oxidative stress, DMSO which is
the solvent to dilute ETST Hemp CBD in media and different
concentrations of ETST Hemp CBD with and without H2O2. Error bar =
Standard deviation. Control Media alone and DMSO alone. Positive
control: H2O2.
When brain cells are incubated with H2O2 by itself lipid
peroxidation occurs substantially as seen in the graph above. Lipid
peroxidation as measured my MDA present is seen to reach greater
than 2.5 nM as compared to controls of media and DMSO alone which
register 0.5 nM or less. When human brain cells are treated with
ETST High Grade CBD (Cannabidiol) in concentrations varying from 1
M to 10 M prior to the addition of H2O2 (prophylactically), human
brain cells are protected against the damaging effects of lipid
peroxidation.
Figure legend:
Earth Science Tech (ETST)
High Grade CBD (Cannabidiol) Oil deters cell death in human brain
cells in vitro. Human brain cells were cultured and treated with
ETST Hemp CBDemp He diluted to concentrations indicated on the
graph for 2 days (1.0 M CBD = 8 ppm CBD rich hemp oil; 2.5 M CBD =
20 ppm CBD rich hemp oil; 5.0 M CBD = 41 ppm CBD rich hemp
oil). The cells were challenged with 25 M (0.85 ppm) H2O2 for
15 hours and stained with neutral red, which stains only live
cells. The amount of Neutral Red is quantified by absorbance
at 540 nm. The experiment is performed in quadruplicates,
with error bars representing standard deviations. Control: Media.
Positive control: Media + DMSO + H2O2.
When human brain cells are incubated with H2O2 cell death occurs
as compared to normal physiological states of growing cells as
measured using neutral red assay (see figure). When ETST Hemp CBD
is used prophylactically at various concentrations, cell death is
deterred even in the presence of H2O2.
DV Biologics Summary
Reactive oxygen species (ROS) and lipid peroxidation is damaging
to human brain cells. ETST High Grade CBD (Cannabidiol) Oil in
varying concentrations has shown to rescue the damaging effects of
ROS and lipid peroxidation, two oxidative stress pathways that play
a crucial role in various neurodegenerative disorders, cancer and
aging. Moreover, ETST High Grade CBD (Cannabidiol) Oil deters cell
death that occurs following the addition of hydrogen peroxide, a
known ROS and lipid peroxidation inducer. This suggests a
neuroprotective effect in vitro to human brain cells following the
use of ETST Hemp CBD. This data merits the use of ETST High Grade
CBD (Cannabidiol) Oil in human clinical studies for
neurodegenerative disorders such as Alzheimers and Parkinsons.
*This is the end of the DV Biologics Study and
Report*
Earth Science Tech (ETST) is
focused on the science, research and studies of its High Grade Hemp
CBD Oil as a nutraceutical and dietary supplement. In
order to assure the public that ETST provides the best High Grade
Hemp CBD (Cannabidiol) Oil, we continue to focus on providing the
public with sound scientific research and keeping the public
informed on the progression of studies being done on our High Grade
Hemp CBD products. Science is what will progress the future of ETST
High Grade Hemp CBD (Cannabidiol) Oil and its products. Cannabidiol
(CBD) is a naturally occurring constituent of industrial hemp oil.
It is the most abundant non-psychoactive cannabinoid in
hemp/cannabis, and is commonly used as a dietary supplement to
support general wellness. ETST all natural full spectrum High Grade
Hemp CBD Oil is derived from Grade-A European Industrial Hemp
Plants. ETST Hemp CBD oils are constantly analyzed for their purity
and purity from potential contaminants. ETST encourage researching
cannabidiol (CBD) from reliable informational sources to see what
is being investigated and discussed about CBD oil. Numerous
properties and benefits of CBD are being investigated at academic
research centers in the United States and elsewhere. The FDA has
not evaluated the validity or truthfulness of these claims;
therefore, we encourage you to review published researches relating
to the benefits and properties of Hemp CBD oils and other Hemp CBD
products.
ETST invites all interested parties to contact the
company at info@earthsciencetech.com in regards
to securing its"High Grade Hemp CBD (Cannabidiol) Oil" in bulk for
wholesale, resale, private labeling or distribution. CBD
(Cannabidiol) is a natural constituent of hemp oil and legal to
consume, sell and possess in the U.S.A. and over 40 countries as
dietary supplement. From product formulating, mixing, labeling and
launching ETST can do it all. ETST is always open to forming new
strategic partnerships, joint ventures and/or supplying vendors,
resellers and third party distribution companies with its High
Grade Hemp CBD (Cannabidiol) Oil/Products at wholesale prices in
order to better help supply consumers as well as the health and
wellness markets on a large scale. You can view, research and
purchase small quantities or bottles of ETST High Grade CBD
(Cannabidiol) Rich Hemp Oil at www.earthsciencetech.com/cbd-rich-hemp-oil.
Humans are designed with a built-in Endocannabinoid
System (ECS). Research shows that many scientists believe
cannabinoids work synergistically to support the whole human
endo-cannabinoid system (internal and natural human cannabinoid
system). It is your body's way of communicating with itself and
properly regulating the way every other body system functions from
your brain to your toes. The Endocannabinoid System (ECS) has shown
to control pain sensation, appetite, temperature regulation, stress
reactivity, immune function, and sleep as well as other processes.
Even more interesting is that muscle and fat tissue also utilize
these receptors to control their processes. Consumers should think
of the Endocannabinoid System as one of the body's main control and
command centers for tweaking your metabolism's ability to react and
adapt to the world around it. Like many other nutrients in our
diet, Cannabinoids can become exhausted and in need of steady
dietary replenishment. Scientific studies and data show that Hemp
CBD (Cannabidiol) Oil as well as all micronutrients
(phytonutrients) and phyto-cannabinoids found in the oil work in
harmony with the human body's own endocannabinoid system making a
positive effect for overall wellness. Hemp oil contains CBD -- a
non-psychoactive compound of the plant. There are millions of hemp
oil consumers around the world and this number is rapidly rising
with an increasing number of reports and studies showing a variety
of benefits of hemp oil.
ETST High Grade Hemp CBD (Cannabidiol) Oil:
ETST High Grade Hemp CBD (Cannabidiol) Oil is all-natural, organic
and formulated using a wide array of cutting-edge technologies.
ETST's High Grade Hemp CBD Oil is all-natural and derived
completely from the federally legal industrial hemp plant.
Industrial Hemp (Hemp) is not marijuana and will not get you 'high'
and it does not require a medical license of any kind to authorize
purchase. It is lab tested multiple times during the manufacturing
process, from seed to shelf. This also includes being tested for
CBD content, other Cannabinoid content, yeast/mold/fungus, and
bacteria like E. coli to ensure safety and top quality. ETST
believes that Hemp CBD Oil awareness today is where Omega 3 and
Omega 6 fatty acids were 15 to 20 years ago. ETST goal is to help
consumers Worldwide with their overall wellness through advanced
formulations with its High Grade Hemp CBD (Cannabidiol) Oil and
other cutting edge nutraceuticals and dietary supplements. Hemp CBD
Oil is posed to be the most renowned and effective natural compound
and botanical alternative introduced to the nutritional and dietary
supplement industry in decades. Consumers worldwide desperately
need more effective safe botanical alternatives to help them with
their overall health and internal wellness. Modern science has
identified over 400 phytonutrients in various parts of the
industrial hemp plant. These include hundreds of Terpenoids,
Essential Oils and Antioxidants but perhaps the most dynamic are a
group of 80 compounds called Cannabinoids that are exclusively
found in Hemp and one other place; our bodies. There are roughly 80
different phyto-cannabinoids, most prominently CBD, CBC, CBG, CBN,
and most likely others yet to be effectively identified. In
addition to the cannabinoids present in High Grade Hemp CBD
(Cannabidiol) Oil extracts, there are also many other types of
natural molecules such as amino acids, fatty acids, flavonoids,
ketones, nitrogenous compounds, alkanes, alcohols, glycosides,
pigments and terpenes among other things. The most common terpenes
in our Hemp CBD oils are Beta-caryophyllene, Caryophyllene Oxide,
Myrcene, Terpinolene, Bisabolol, Pinene, Nerolidol, Ocimene and
Phytol.
ABOUT ETST (www.earthsciencetech.com): ETST (ETST) is a
unique Biotechnology company focused on delivering cutting edge
Nutraceuticals, Bioceuticals and Dietary Supplements designed for
excellence in the areas of Health, Wellness, Sports and Alternative
Medicine and to improving the quality of life for Consumers
Worldwide. ETST is also dedicated to providing Natural Alternatives
to prescription medications through the use of its cutting edge
Nutritional and Dietary Supplements. This may include products such
as its High Grade Hemp CBD (Cannabidiol) Oil, Vitamins, Minerals,
Herbs, Botanicals, Personal Care Products, Homeopathies, Functional
Foods and other products. These products may be in various
formulations and delivery systems including (but not limited to)
capsules, tablets, soft gels, chewables, liquids, creams, sprays,
powders, and whole herbs. ETST is focused on researching and
developing innovative Hemp extracts and to make them accessible
Worldwide. ETST plans to be the premier supplier of the highest
quality and purity of High Grade Hemp CBD (Cannabidiol) Oil. ETST's
primary goal is to advance different High Quality Hemp extracts
with a broad profile of Cannabinoids and additional natural
molecules found in Industrial Hemp and to identify their distinct
properties. The company is dedicated in offering its consumers the
finest and purest quality All Natural-Organic Hemp CBD Oil while
never compromising on quality. ETST High Grade Hemp CBD
(Cannabidiol) Oil is classified as "food based" and therefore
perfectly permissible in all 50 states and more than 40 countries.
Cannabinoids (Cannabidiol/CBD) are natural constituents of the Hemp
plant and CBD is derived from Hemp stalk and seed. Hemp oil is a
well-known dietary supplement and the naturally occurring CBD
possesses no psychoactive qualities and presents a continuing
stream of overwhelming evidence of significant Wellness benefits.
With no psychoactive ingredient, Hemp CBD Oil is a ready-for-market
Hemp-based Nutraceutical. The United States Food and Drug
Administration (FDA) currently considers non-THC hemp based
cannabinoids, including CBD, to be "food based" and therefore
saleable. These new non-psychoactive CBD-rich hemp oil products
that ETST has geared up to market and distribute are beyond
reproach. CBD (cannabidiol), a naturally occurring constituent of
the Industrial Hemp plant, promotes and supports the nutritional
health of aging bodies in particular. Source: US Government Patent
#6,630,507 "Cannabinoids as antioxidants and neuroprotectants."
ETST does not grow, sell or distribute any substances that violate
United States Law or the controlled substance act. ETST does sell
and distribute hemp based products.
ABOUT DV BIOLOGICS (www.dvbiologics.com):
DV Biologics is a global supplier of human biological tools to
academic institutions and pharmaceutical companies engaging in cell
and drug based discovery and development. Our mission is to provide
biological tools needed to advance the innovation of technology
that will ultimately be used to treat or prevent multiple human
degenerative disorders and diseases. DV Biologics offers a diverse
range of novel human biological tools and services that can be used
to study various human pathological conditions in addition to an
expanded product portfolio of unique cell types and tissue-derived
products.
FOOD AND DRUG ADMINISTRATION (FDA) DISCLOSURE:
These statements and products have not been evaluated by the FDA
and are not intended to diagnose, treat or cure any disease. Always
check with your physician before starting a new dietary supplement
program.
FORWARD LOOKING DISCLAIMER: This release
contains forward-looking statements that involve risks and
uncertainties. Readers are referred to the Securities and Exchange
Commission filings filed by the Company on EDGAR at http://www.sec.gov/edgar.shtml,
specifically the most recent reports which identify important risk
facts that could cause actual results to differ from those
contained in the forward-looking statements. The Company undertakes
no obligation to review or confirm analysts' expectations or
estimates or to publicly release any revisions to any
forward-looking statements. The information contained in this press
release should not be construed as any indication of the Company's
future stock price, its revenues or results of operations.
Contact:
Earth Science Tech, Inc. (ETST)
Boca Raton, FL
info@earthsciencetech.com
Phone: +1(305) 615-2118
DV Biologics
Biotechnology Company
Costa Mesa, CA