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Relation of antistress and geroprotective effects of deuterium depleted water in aging female rats

Authors:
Issue 10. May 2017 | Cardiometry | 35
DOI: 10.12710/cardiometry.2017.3542
Relation of antistress and geroprotective effects of
deuterium depleted water in aging female rats
Oleg I. Kit1, Alla I. Shikhliarova1, Galina V. Zhukova1*, Stepan S. Dzhimak2,
Mikhail G. Barishev2, Tatiana A. Kurkina1, Elena A. Shirnina1, Tatiana P. Protasova1
1 Rostov Research Institute of Oncology
Russia, 344037, Rostov-on-Don, 14th line str. 63
2 Kuban State University
Russia, 350040, Krasnodar, Stavropolskaya str. 149
* Corresponding author: e-mail: galya_57@mail.ru
Abstract
The use of deuterium depleted water (DDW) in a dosing regimen of 25–30 ml per day
for five weeks in aging female rats (20–22 months) contributed to the overall correc-
tion of age-related disorders of the adaptive status, estrous cycle regulation and non-
specific resistance, as well as the appearance of visually distinct signs of rejuvenation of
the animals. The obtained results show the relation between the geroprotective effect
of DDW and the development of the general nonspecific antistress adaptation reac-
tions of calm and elevated activation that can serve as an experimental basis for the
development of an effective method of geriatrics and prevention of cardiovascular dis-
eases and degenerative processes in women in the peri- and postmenopausal period.
Keywords
Deuterium depleted water, Aging female rats, Estrous cycle, General nonspecific
antistress adaptation reactions, Neuroendocrine regulation
Imprint
Oleg I. Kit, Alla I. Shikhliarova, Galina V. Zhukova, Stepan S. Dzhimak, Mikhail G. Barishev,
Tatiana A. Kurkina, Elena A. Shirnina, Tatiana P. Protasova. Relation of antistress and
geroprotective effects of deuterium depleted water in aging female rats. Cardiometry;
No.10 May 2017; p.35–42; DOI: 10.12710/cardiometry.2017.3542; Available on-
line: http://www.cardiometry.net/issues/no10-may-2017/geroprotective-effects
ORIGINAL RESEARCH
Introduction
Age-linked disorders of the neuro-
endocrine and immune regulation,
as a rule, lead to a reduction in the
nonspecific resistance of an organism
and are responsible for development
of various pathological states, which
may include cardiovascular, degener-
ative and tumor-related diseases [1,
11, 21]. In addition, there are some
specific sex-linked differences in the
age-associated disorders, which man-
ifest themselves as a faster decline in
the reproductive function in the fe-
male organism in comparison to the
male one, along with the existence of
a certain relation between the hor-
monal function of the ovaries and
the resistance of the female organism
to cardiovascular and some other
diseases [19, 28]. It has been demon-
strated that, at the level of the entire
organism, the key role in providing
aging-retarding conditions should
be assigned to a mechanism respon-
sible for generation and changes of
some discrete states in the organism,
namely, some integral adaptation re-
actions of stress [20] and antistress
[3, 4] in character. An accumulation
and predominance of aging-related
stress alterations provoke a gradual
decline in the nonspecific resistance.
In contrast, the development of sta-
ble reactions of the antistress type
contributes to activation of CNS, the
endocrine and immune systems, the
energetic and peroxidation metabo-
lism, i.e. the most important compo-
nents governing an elevation of the
organism resistance to unfavorable
external and internal environment
factors.
Progression of age-associated dis-
orders depend on many of exoge-
nous and endogenous factors, among
them the peculiarities of the water
condition profile in an organism that
is of vital importance for biochem-
ical processes and realization of ef-
fects produced by weak information
actions of water due to the unique
cluster structure of water and its
metastability [18, 26]. In this case, it
should be mentioned that known are
some biotropic properties of water,
which are determined by its actual
isotope composition. It has been de-
tected that even some slight changes
in the relationship between the stable
hydrogen isotope ratios may have
a noticeable effect on the states of a
living organism. By this means it has
been found that water enriched with
the deuterium is quite often the cause
of some pathological changes in the
Submitted: 19.2.2017, Accepted: 10.3.2017, Published online: 25.5.2017
36 | Cardiometry | Issue 10. May 2017
organism; in case when we deal with
a shift in the water isotopic compo-
sition to light protium water, on the
contrary, this type of water is capable
of improving the functional state of
a human or an animal [24, 25, 27].
A number of favorable effects pro-
duced by deuterium depleted water
(DDW) on the conditions of various
protection systems in the organism
have been detected [7, 17, 23], up to
and including pronounced triggering
action by DDW on the mechanisms
of the anti-tumor resistance [16, 22].
Evidence data obtained from epide-
miological studies show that there
are cases of extreme longevity for
those individuals reported who drink
original water supplied by some nat-
ural springs, differing from standard
drinking water in a number of its
properties, among which is deuteri-
um depletion [15]. All this allows us
to consider DDW use as a new way
for correction of some homeostatic
disorders associated with aging.
Our challenge was to carry out ex-
perimental studies addressing a cor-
rective effect produced by DDW on
the adaptation status, the state of the
estrous cycle and the microflora com-
position on skin surfaces in outbred
female Albino rats in their aging pe-
riod.
Materials and methods
Our experimental studies were per-
formed in the early spring period in
27 outbred Albino aging female rats
aged 20–22 months, who had weights
290,7 ± 31,6 g, against 10 young fe-
male rates aged 8–10 months with a
weight of 210,4 ± 31,6 g. All our ex-
perimental studies involving use of
animals have been carried out in full
compliance with the applicable inter-
national bioethics rules, guidelines
and regulations [12]. The aging an-
imals were divided into two groups:
Group A and Group B, correspond-
ingly. Group A included 13 rats who
received DDW with a daily amount of
25–30 ml during 5 consecutive weeks.
Group B covered 15 female rats who
received only standard drinking wa-
ter, the quality of which met the re-
quirements specified by the relevant
Russian National Standard GOST.
DDW had a deuterium concentra-
tion three times lower than it was the
case with regional standard-type nat-
ural water (150 ± 6 ppm) and showed
no essential differences from the lat-
ter in its mineral analysis.
In order to assess effects induced by
DDW on the adaptation status of the
animals, in our experimental studies
traced was dynamics of progression
of general nonspecific adaptation re-
actions (GNARs) of the organism [3.
4]. Each type of GNARs, covering the
reactions of acute stress and chronic
stress as well as the antistress reactions
of training, calm and elevated activa-
tion, was identified according to the
Shilling’s blood count data with the
use of Giemsa – Romanovsky staining
technique, with a 200 blood cell count-
ing accuracy. In doing so, the percent-
age of lymphocytes in peripheral blood
was taken as an identifier making pos-
sible to properly attribute each actual-
ly initiated reaction to the respective
GNAR type. The corresponding leu-
kogram patterns in the animals were
identified first initially, prior to the
experimental studies, and later, on a
regular weekly basis, in the course of
the experiment. Blood samples were
taken from a rodent femoral vein lo-
cated under the skin in the cellular tis-
sue, throughout the medial surface, in
the morning time before feeding. For
the purpose of processing of the exper-
imental data, utilized was our original
Software Statistica 6.0, and the statisti-
cal analyses were completed in accor-
dance with Students t-test technique.
Cytological data on the functional
status of the ovaries were obtained
using vaginal smear reading. Upon
flushing from the vaginal lining,
smears were read with the Leica DM
LS2 microscope two times a day, in
the morning and in the evening, re-
spectively. For that purpose, the fol-
lowing phases of the estrous cycle
were traced: 1. Diestrus (D) phase
which implies the functional rest and
which is marked by the presence of
leu-kocytes and mucus found in the
smears; 2. Proestrus (Р) phase recog-
nized by the presence of polygonal,
usu-ally nucleated, epithelial cells,
with excentrically located nuclei; 3.
Estrus phase (О) when non-nucleat-
ed cornified squamous epithelial cells
appear and form large clumps and
sheets; 4. Metestrus phase (М), the
day after estrus, exhibiting a mixed
cell composition, when a mixture of
cornified squamous epithelial cells
and leukocytes predominates. Length
of each phase or stage and their se-
quences within the estrous cycle were
recognized that has made possible
to evaluate periodicity of hormonal
changes in the ovaries [2].
In order to assess the state of the
nonspecific resistance of the organ-
ism, under administering DDW to
aging female rats, applied has been the
trivial assessment procedure by Klem-
parskaya – Shalnova for the proper
identification of autoflora species on
the skin as current status testing for
information [6]. To examine the actual
nomenclature of the microorganisms
Issue 10. May 2017 | Cardiometry | 37
sampled from rat tails with the agar
replica test technique, the Korostylev
medium consisting of meat-pepto-
ne agar with an addition of a 1,5%
Bromthymolblau alcohol solution and
a 1% mannit solution was used. The
cultivated bacteria have been differ-
entiated and recognized according to
their ability to ferment mannit and
change color of the colonies of Staph-
ylococci both of pathogenic and non-
pathogenic nature.
Results and discussion
Our analysis of leukocyte count data
on peripheral blood in aging rates in
the above mentioned Groups com-
pleted first before our experimental
studies has revealed that the animals
in 88,6 % of cases had low relative
numbers of lymphocytes (30–45 %)
at a rather high total amount of leu-
kocytes in blood (see Table 1 below
herein). Some cases of aneosinophilia
were reported, too.
The said indices were in correspon-
dence to the leukogram pattern typi-
cal for the symptom complex of stress.
Moreover, they were statistically sig-
nificantly different from the hemato-
logical characteristics in the female
rats at a young age, whose lympho-
cyte percentage was 1.7 times greater
Table 1. Leukogram data as percentages of major blood cells in aging female rats under DDW influence as against their initial state and
as against the state of the reference group of young and aging female rats without DDW administration
Blood cell types Young female rats
Aging female rats
Initial state After DDW administration Reference
Group А: activation Group А: training Group В: stress
Total leukocyte count /mcL 7300±1200 17100±2900 19100±1700 210800±1200 216500±1700 1
Lymphocytes, % 75,5±4,5 44,0±2,94 168,0±2,45 256,5±1,7 237,5±2,15 1
Eosinophils, % 1,0±0,13 0,5±0,11 1,0±0,7 1,5±0,1 0
Neutrophils, % 27,5±3,5 42,0±2,7 130,0±3,3 246,0±2,9 62,0±4,4 1
Notes:
1 statistically significantly differing from the values in young female rats (p<0,001);
2 significantly differing from the values in aging female rats in their initial state (p<0,001).
as compared with the aging rats, with
a lower total leukocyte count and
with no cases of aneosinophilia.
Upon completion of the DDW treat-
ment course, the animals in Group A
demonstrated the normalized leuko-
gram pattern. First it was applicable
to the relative number of lymphocytes
as the key index, showing the type of
the adaptation reaction of the organ-
ism, which was higher by a factor of
1,5 in comparison with the initial state
(see Table 1 below herein). The total
count of leukocytes in blood became
normal, too, and no aneosinophilia
cases were observed. The peripheral
blood leukogram changes in question
made it possible to recognize the de-
velopment of the integral reactions of
the physiological classes in the Group
A rats as follows: GNARs of training,
calm and elevated activation. The he-
matological data obtained from the
animals in Group B, who received
standard drinking water, practical-
ly did not show any differences from
their initial values: an elevated total
count of leukocytes was recorded, and
lower lymphocyte percentages were
reported than those identified in the
young animals. So, an analysis of de-
veloping GNARs of differing types
within each Group and between the
specified Groups has shown that the
use of DDW instead of the standard
drinking water has resulted in a sub-
stantial increase in the occurrence
rate of the antistress reactions detect-
ed in Group A covering the aging fe-
male rats.
Table 2 below herein summarizes
variability in the occurrence rate of
different GNARs for a five week peri-
od in aging female rats of the Groups
under examination. As may be seen
from the Table, the occurrence rate
of the stress reaction in Group A
identified prior to the administration
of DDW was comparable with that
reported for Group B, and the occur-
rence rate of the antistress reactions
of different types has varied. Upon
DDW receiving, during the first
week, reported were GNARs of the
antistress types in the Group A rats
only, and the same stable maintenance
of the physiological reactions was re-
corded till the completion of the ex-
perimental studies. In the animals of
Group B, in 35% of cases, in contrast
thereto, observed was stabilization of
the development of the stress reac-
tion pattern, which was especially ap-
parent by the end of the experiment.
Differences of decisive importance
between the aging animals belonging
38 | Cardiometry | Issue 10. May 2017
to different Groups were also found in
the occurrence rates of the reactions
of the same name of the antistress
types. So, beginning with week 2 till
week 5 of the experiment time, much
more frequently (up to 3–7 times) ob-
served were the reactions of the calm
and elevated activation in Group A,
which were considered to be the most
favorable for the neuroendocrine reg-
ulation, against those seen in Group B,
where and when the reaction of stress
was accompanied by the antistress re-
action of training.
The established inter- and intra-group
relations between the antistress (AS)
and stress (S) reactions have demon-
strated the dominance of the antistress
link within the entire structure of the
adaptation reactions in the animals,
who received DDW, and actually it has
shown high effectiveness of the DDW
action. As is known, the calm activa-
tion GNAR and the elevated activation
GNAR are featured by higher per-
centages of lymphocytes in blood (for
outbred rats 61–70% and 71–80%,
correspondingly), in comparison with
the stress GNAR and that of training
(under 50% and under 51–60%, cor-
respondingly). The above performed
correlation & regression analysis of
the relationship between the level of
lymphocytes and the respective mor-
Table 2. Relations of occurrence rates of different adaptation reactions in aging female rats receiving DDW (Grpup A) vs. those receiving
standard drinking water (Group В)
Types of adaptation reactions
Relation of occurrence rates of ARs of the same name (%)
in aging animal groups under examination (A vs. B)
Initially Week 1 Week 2 Week 3 Week 4 Week 5
Stress 20,3/21,4 0/21,4 0/21,4 0/35,7 0/35,7 0/35,7
Training 44,1/40,8 30,8/31,4 30,8/57,2 15,4/50,0 15,2/50,0 0/50,0
Calm activation 20,6/20,6 38,4/28,6 23,1/21,4 38,4/14,3 30,8/14,3 30,8/0
Elevated activation 15,0/17,8 30,8/18,6 46,1/0 46,2/0 54,0/0 69,2/14,3
Intra-group relation between ARs:
Antistress / Stress reation in Group A 3,9 AS only AS only AS only AS only AS only
Intra-group relation between ARs:
Antistress / Stress reation in Group B 3,7 3,7 3,7 1,8 1,8 1,8
phological, biochemical and hormonal
indices of the functional state of the
nervous, endocrine and immune sys-
tems in the organism has revealed that
we deal with high values of the paired
and multiple correlation coefficients
as well the determination, which are
decisive for production of a strong cor-
relation between the key index and the
multisystem attributes of an adaptation
reaction [9, 10].
As is seen, the mechanism of devel-
opment of the antistress calm and ele-
vated activation GNARs has contrib-
uted to a favorable correction of the
length of the estrous cycle stages in
the rodents. The final data obtained
in the Groups in question by the end
of the experimental studies should be
treated as evidence in support of the
DDW effect on the organism for the
aging female rates in Group A. As is
indicated in Table 3, the estrous cycle
length in the aging animals in Group
B, who received standard drinking
water, differed markedly from that
recorded in the young animals in the
reproduction age and was featured
by a considerably reduced length of
the proestrus and estrus stages (2-3
times) with a significantly increased
length both of the metestrus phase
(more than 3 times) and dioestrus
(more than 1.5 times) stage. The
DDW use for five consecutive weeks
resulted in the statistically signifi-
cant changes found in the estrous
cycle that should be considered as
evidence for the partial restoration
of the reproductive function in the
aging female rats in Group A. So, by
the end of the experiment, the ani-
mals in Group A have demonstrated
a substantial shortening of the length
of the metestrus and dioestrus stages
(1,5 and 1,8 times, respectively) and
a lengthening of the proestrus stage,
as compared with Group B, that has
placed the said indices closer to those
recorded for the young age rats (see
Table 3 below herein).
When comparing the length of the
individual stages in common within
the estrous cycle over the entire peri-
od of observations, under developing
of GNARs of various types, a clear-
cut relationship between the integral
changes and the hormonal fluctua-
tions was also traced. It appeared as
the restored length and sequence of
the individual stages within the cy-
cle that was primarily detected under
formation of the stable reactions of
activation under the DDW effect. As
evidenced by the data given in Table 4,
when supporting GNARs of the calm
and elevated activation in the Group
A rats, the length of each individual
Issue 10. May 2017 | Cardiometry | 39
Table 3. DDW administration effect on the length of individual stages of estrous cycle in aging female rats
Animal Groups Length of a separate stage of the estrous cycle (hours)
DР О М
Norms for young age 57 (55–57) 12 (12–14) 27 (25–27) 6 (6–8)
Aging female rats Group В 91,1±7,0 4,1±0,7 15,0±2,4 25,5±3,8
Group А 51,2±5,4 19,1±1,8 113,8±2,7 16,5±2,4 2
Note:
1 statistically significant differences from values in Group В, p < 0,01–0,05
stage of the cycle in common, con-
sidering a period of 30-35 days, was
brought closer to the normal indica-
tive values, that is to say, to those val-
ues identified in the young rat Group.
So, the length of the diestrus stage in
general in those cases decreased ap-
proximately by 50 hours in compari-
son to the same value in Group B, and
it was found to be maximally close
to the values detected in the young
age animals. The duration of the
proestrous stage in the Group A rats,
with the development of GNAR of the
calm and elevated activation, demon-
strated no difference from the normal
values. At the same time, in the ani-
mals in Group B, with the prevalence
of GNAR of training, when and where
the reaction of stress was often record-
ed, the length of the above mentioned
stage was 3 times shorter than that re-
ported for the young female rats.
The restoration of the estrus stage,
which either has been absent in the
initial state or exceeded by 1.6 times
the limits of the stage range in young
female rats, is of considerable impor-
tance for an evaluation of the gero-
protective effect produced by DDW
with the development of GNAR of
calm and elevated activation. The du-
ration of the metestrus stage in female
rats in group A exceeded the norm,
but was recorded to be statistically
significantly shorter than that under
the development of the training reac-
tion. In cases of development of the
reaction of the same name in animals
in group B, the process of the estrous
cycle normalization was moderate-
ly expressed only, but the indices of
change of the phases and their length
in common differed from those re-
corded under stress, which was char-
acterized by profound disorders in
Table 4. Change in length of individual stages within the estrous cycle in general during the experiment period (in hours) in aging
female rats with the development of different GNARs under DDW effect
Stages of estrous cycle Young female rats
Aging female rats
Group В Group А
GNAR of training and
stress
GNAR of calm and
elevated activation GNAR of
Diestrus 346,6±7,2 389,4±42,2 training 316,8±19,9
Proestrus 71±2,4 24,1±1,7 178,1±2,4 260,3±7,9 1,2
Estrus 148±14,7 235,1±39,1 1144,7±11,3 264,5±7,6 1,2
Metestrus 32,4±2,2 20,5±3,8 147,4±2,3 1,2 58,4±2,7 1,2
Notes:
1 statistically significantly differing from values in young rats (р<0,01)
2 statistically significantly differing from values in aging rats (р<0,01)
the sexual function rhythmic activity
in aging female rats in the reference
group under examination.
Microscopic examination of vaginal
smears in the aging female rats pro-
vided a means for visualizing the reg-
ulatory effect by DDW at the cell level.
Figure 1 below herein exhibits vaginal
smear readings obtained in the aging
female rats who received DDW, where
the cell composition corresponds to
the normal cell types of the estrous
cycle stages. Epithelial cells prevailed
in the proestrus phase; squamous ep-
ithelial cells dominated in the estrus
phase; mixed composition of cells
(cornified squamous cells, epithelial
cells, leukocytes) were dominant in
the metestrus phase, and leucocytes
were prevalent in the diestrus stage.
An interpretation of the obtained
results from the standpoint of the es-
trous cycle neuroendocrine regulation
40 | Cardiometry | Issue 10. May 2017
in the context of the dominant inte-
gral reaction of activation, under the
DDW effect, has demonstrated that
the stimulation by estrogens, which
are produced by ovaries and initiate
the vaginal proliferative activity, de-
termines also the cell composition.
An indication thereof firstly are the
implied DDW effects responsible for
approaching the normal value of the
proestrus stage, that indirectly con-
firms the fact of the activation of hy-
pophysis follicle-stimulating hormone
secretion. Secondly, despite the fact of
the normalization of the estrus pe-
riod, it has become apparent that we
deal with a decrease in the metestrus
period length with reference to the
respective values in the aging female
rats as an evidence of the corpus lute-
um shrink and inhibition of excessive
luteinizing hormone secretion. And,
thirdly, the noted normalization of the
metestrus stage has shown there has
Figure 1. Cell composition of vaginal smears read at different stages of estrus cycle in aging
female rats under DDW influence:
1) proestrus; 2) estrus; 3) metestrus; 4) diestrus.
been found the state of hormonal bal-
ance approaching.
The studies identifying the skin aut-
oflora peculiarities in aging female
rats have also revealed some differ-
ences attributed to the type of evolving
GNARs by comparing two cases: that
with DDW administration and anoth-
er one with use of standard drinking
water (see Table 5 below herein).
Thus, with the development of the
stable calm and elevated activation
reactions, under the DDW effect, ob-
served has been a noticeable suppres-
sion of growth up to even complete
disappearance of pathogenic staphylo-
coccus with a negative mannit effect of
the “yellow” colonies formation, while
the “green” colonies with a positive
mannit effect of nonpathogenic forms
of staphylococcus remained. With the
development of the training reaction,
the “green” colonies significantly pre-
vailed over the “yellow” ones. Under
stress, in animals of Group В observed
has been an increased pathogenicity in
skin autoflora: against the background
of suppression of the “green” colonies,
a progressive growth in the «yellow»
colonies of pathogenic staphylococcus
has been detected. Hence, the domi-
nant in Group A adaptation reactions
of calm and elevated activation are the
most efficient, when considering the
antimicrobial resistance.
The antistress effect induced by the
DDW administration led to an ap-
pearance of visually distinguishable
signs of rejuvenation in the Group A
animals. In the female rats, receiving
DDW, a change in hair coat was re-
corded. Observed was replacement of
a coarse stiff hair coat, yellowish toned,
comprising multiple local areas char-
acterized by varying levels of hair loss,
by a white soft fur with a new thick un-
dercoat; the eye sclera have brightened
(see Figure 2), and the motion activity
significantly higher in the animals un-
der examination was also reported.
Thus, the obtained data demonstrate
pronounced geroprotective DDW ef-
fect associated with DDW antistress
action which has manifested in for-
mation of stable GNAR of the calm
and elevated activation, contributing
to a considerable increase in nonspe-
cific resistance and initiation of the
restoration process in the aging fe-
male rats. It is obvious that the said
effects take place due to such a specif-
ic property of DDW as its capability
to activate the cell metabolism and
elevate excitability of the neuroendo-
crine regulation centers [13, 14].
The data on the development of
GNAR of the calm and increased ac-
tivation under the DDW influence are
in good agreement with the known
findings in favor of the activating effect
Issue 10. May 2017 | Cardiometry | 41
Table 5. Changes in skin autoflora in aging female rats under DDW effect, depending on
the adaptation reaction type
Types of microflora
colonies
Group В
GNAR of stress
Group A
GNAR of calm and
elevated activation GNAR of training
Total number of the
colonies 43,6±2,18 57,0±1,6 134,3±1,27 1
“Yellow” colonies
– pathogenic
staphylococcus
36,9±5,1 N/A 5,9±1,3 1
«Green» colonies
– nonpathogenic
staphylococcus
7,0±1,3 54,5±3,05 126,3±2,8 1
Note:
1 statistically significantly differing from values in group B, р<0,01
Figure 2. Change in appearance of aging fe-
male rats 5 weeks after DDW administration.
1. Group B: Female rats receiving drinking
water.
2. Group A: Female rats receiving DDW.
made by DDW on the CNS serotoner-
gic structures [25], since the relation-
ship between the given GNAR devel-
opment and an increase in serotonin
concentration in brain structures and
blood formed elements had been es-
tablished earlier [5]. This fact is of great
significance due to an important role
of stress-limiting serotonergic process-
es for the purpose of ischemic heart
damage prevention and a reduction of
cardiovascular disease risks [7] related
to pathologies associated with aging.
The evidence of the correlation be-
tween mortality due to cardiovascular
and degenerative diseases and a decline
in the reproductive function in females
during peri- and postmenopause re-
veal an additional aspect in the import-
ant role of properly maintaining the
ovarian hormonal function to provide
resistance of the female organism to
damaging factors [28]. In this connec-
tion, the antistress and geroprotective
DDW effects detected and traced in
our studies is a matter of great interest.
Conclusions
The results obtained in our experi-
mental studies bear witness that there
is pronounced geroprotective effect
produced by DDW in outbred aging
female rats and that there exists the
relation between this effect and the
development of the general nonspe-
cific antistress adaptation reactions of
calm and elevated activation. The ob-
tained evidence data provide unique
insight into the effect exerted by DDW
on mammalian organisms, and it may
serve as an experimental foundation
for development of an effective meth-
od suitable for use in geriatrics and
prevention of cardiovascular diseases
and degenerative processes in females
during peri- and postmenopause.
Statement on ethical issues
Research involving people and/or ani-
mals is in full compliance with cur-
rent national and international ethi-
cal standards.
Conflict of interest
None declared.
Author contributions
The authors read the ICMJE criteria
for authorship and approved the final
manuscript.
References
1. Anisimov VN. Molecular and
physiological mechanisms of aging.
St. Petersburg. 2008. V.1 461 p. [in
Russian]
2. Volkova OV. Structure and regu-
lation of ovary function. Мoscow:
Meditsina, 1970. 68 p. [in Russian]
3. Garkavi LKh, Ukolova MA, Kvak-
ina EB. Law of development of qual-
itatively different general unspecific
adaptation reactions. Patent No.158,
Bulletin of Discoveries in USSR.
Мoscow, 1975. No. 3. p. 56–61. [in
Russian]
4. Garkavi LKh, Kvakina EB, Kuzmen-
ko TS, Shikhliarova AI. Anti–stressor
reactions and activation therapy.
Ekaterinburg, 2002. Part I. 196 p. [in
Russian]
5. Zhukova GV, Garkavi LKh, Mikhai-
lov NU, et al. On information value
of some histochemical, cytologic and
biorhythmical indicators for evalua-
tion of the organism functional state.
42 | Cardiometry | Issue 10. May 2017
Bulletin of Southern Scientific Cen-
ter of Russian Academy of Sciences.
2010;6(3):49–59. [in Russian]
6. Klemparskaya NN, Shalnova GA.
Autoflora as an indicator of radiation
affection. Moscow : Meditsina, 1966.
208 p. [in Russian]
7. Meerson FZ, Pshennikova MG.
Adaptation to stress situations and
physical loads. Мoscow, 1988. [in
Russian]
8. Timakov AA, Smirnov VM, Gusa-
rov EA. Immunity increase by natural
way. М.: Nauka, 2003, 51p. [in Russian]
9. Shikhliarova AI. Correction of in-
tersystem relations on experimental
model of tumor growth. Mechanisms
of integration in biological systems.
Problems of adaptation. Stavropol,
1989. p. 67–70. [in Russian]
10. Shikhliarova AI. On possibility
of predicting the efficiency of DDW
influence on living systems. Modern
problems in research and preserva-
tion of biosphere. Saint Petersburg
:Gidrometeoizdat, 1992. p. 179–182.
[in Russian]
11. Dilman V., Dean W. The Neu-
roendocrine Theory of Aging, The
Center for Bio–Gerontology, Pensa-
cola, 1992.
12. European Treaty Series – No. 123.
European Convention for the pro-
tection of vertebrate animals used
for experimental and other scientific
purposes. Strasbourg, 1986
13. Fernandes de Lima V.M., Hanke
W. Modulation of CNS excitability by
water movement. The D2O effects on
the non–linear neuron-glial dynam-
ics. Journal of Biophysical Chemistry.
2011; 2(3):353–60.
14. Goncharuk V, Lapshin VB, Bur-
deinaya TN, et al. Physicochemical
Properties and Biological Activity
of the Water Depleted of Heavy Iso-
topes. X Journal of Water Chemistry
and Technology. 2011;33(1):8–13.
15. Ignatov I., Mosin O. Water in the
Human Body is Informational Bearer
about Longevity. 2012.
16. Krempels K, Somlyai I, Gyöngyi
Z, Ember I, Balog K, Abonyi O and
Somlyai G. A retrospective study of
survival in breast cancer patients
undergoing deuterium depletion in
addition to conventional therapies. J
Cancer Res Ther 2013, 1(8):194–200
17. Lisicin AB, Barishev MG, Basov
AA, et al. Influence of deuterium
depleted water on the organism of
laboratory animals in various func-
tional conditions of nonspecific pro-
tective systems. Biophysics. 2014;
59(4):620–7.
18. Lobyshev V.I. Water is a sensor
to weak forces including electromag-
netic fields of low intensity. Elec-
tromagnetic Biology and Medicine.
2005; 24(3): 449–61. DOI: 10.1080/
15368370500382248
19. Sampson N., Untergasser G., Plas
E., Berger P. The ageing male repro-
ductive tract 2007
20. Selye H. Thymus and adrenals in
the response of the organisms to in-
juries and intoxication Brit. J. Exp.
Path. 1936;17:234–48.
21. Shaw AC, Daniel R, Goldstein
DR, et al. Age-dependent dysregula-
tion of innate immunity. Nature Re-
views Immunology. 2013;13:875–87.
doi:10.1038/nri35472013
22. Somlyai G. Defeating Cancer: The
Biological Effect of Deuterium Deple-
tion. Budapest: AuthorHouse. 2002. 160 p.
23. Somlyai G. Use of deuterium de-
pleted water for the treatment of insulin
resistance. Patent WO2012004620A2.
2012. Global patent index EP 2590715
A2 20130515
24. Somlyai G, Kovács A, Guller I. et
al. Deuterium has a key role in tumour
development – new target in antican-
cer drug development. European Jour-
nal of Cancer. 2010; 8(5): 155–225.
25. Strekalova T, Evans M, Cherno-
piatko A, et al. Deuterium content
of water increases depression sus-
ceptibility: The potential role of a
serotonin–related mechanism. Be-
hav. Brain Res. 2015;277:237–44. doi:
10.1016/ j.bbr.2014.07.039
26. Tsai CJ, Jordan KD. Theoretical
Study of the (H20)6 Cluster. Chemical
Physics Letters. 1993;213:181–8.
27. Vertes A. Physiological effects of
heavy water. Elements and isotopes:
formation, transfor–mation, distribu-
tion. Dordrecht: Kluwer Acad. Publ.,
2004. 112 p.
28. Yang Y, Kozloski M. Sex Differenc-
es in Age Trajectories of Physiological
Dysregulation: In–flammation, Meta-
bolic Syndrome, and Allostatic Load.
Journal of Gerontology: Biological
Sciences. 2011;66(5):493–500.
... Also, the recent developments have demonstrated that reduction of deuterium concentration in organism fluids promotes activation of adaptogenic processes at the molecular level [17][18][19][20][21]. As supposed mechanisms of action of low concentrations of deuterium in drinking diet for an organism, the potentiation of nonspecific defense systems is assumed as well as implementation of "isotopic shock" that leads to the activation of energy reserves accumulation in mitochondria [22,23]. ...
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