The study was performed to assess the impact of Biofield Treated test
item (DMEM medium) on Human Endothelial Hybrid
Cell Line (EA. hy926) for the expression of cyclic guanosine
monophosphate (cGMP). The test item was divided into three parts.
The first part was received one-time Consciousness Energy Treatment by a
renowned Biofield Energy Healer, Mahendra Kumar
Trivedi and labeled as BT-I, while second part received two-times
Biofield Treatment and is denoted as BT-II. The third part did
not receive any types of treatment and denoted as untreated DMEM. The
level of intracellular cGMP in the BT-I and BT-II groups
showed a significantly (p≤0.001) increased by 296.06% and 339.37%,
respectively in Ea. hy926 cells compared to the untreated
DMEM group. These results suggest that BT-II group showed a significant
improved the level of cGMP with respect to the BT-I group.
Therefore, the Biofield Energy Healing Treatment can be used to treat
the erectile dysfunction patients along with other associated
disorders such as orgasmic disorders, frotteuristic disorder, female
sexual arousal disorder, vaginismus, fetishistic disorder, sex
addiction, hypoactive sexual desire disorder, premature or delayed
ejaculation.
Keywords: Biofield Energy; cGMP; Endothelial Hybrid Cell; PDE-5; DMEM; Erectile Dysfunction
Abbrevations:BT-I: One-time Biofield Energy Treated DMEM; BT-II: Two-times Biofield Energy Treated DMEM; CAM:
Complementary and Alternative Medicine; NCCAM: National Center for Complementary and Alternative Medicine; DMEM:
Dulbecco’s Modified Eagle’s Medium; cGMP: Cyclic guanosine 3′,5′-monophosphate; FBS: Fetal Bovine Serum
Introduction
Erectile dysfunction (ED) or impotence is the inability to get
and keep an erection firm enough for sex. This is the most common
sexual disorder in men across the globe [1]. Erectile dysfunction
symptoms include persistent trouble in getting and keeping an
erection and less sexual desire. Male sexual arousal is a complex
process, which includes the coordination of the brain, hormones,
emotions, nerves, muscles, and blood vessels [2]. ED might occur
due to stress and severe mental health conditions. Besides, the
literature data reported that physical cause of ED are heart disease,
Parkinson’s disease, multiple sclerosis, atherosclerosis, high
cholesterol, peronei’s disease high blood pressure, diabetes, obesity,
sleep disorders, use of tobacco, alcoholism, injury of spinal cord
and various metabolic syndromes such as high insulin levels, body
fat around the waist, etc. [3]. Along with physical and psychological
factors, impaired function of arteries and corpora cavernosa within
the penis are the primary condition for impotence. While the lack of
smooth muscle tone and imperfections in neuronal stimuli can lead
to unsuccessful penile erection [4]. ED eventually leads to neuronal
and cardiovascular disorders [5,6].
Nitric oxide synthase (NOS) enzymes are the major mechanism
involved in ED, and it enhanced the production of cyclic guanosine
monophosphate (cGMP), which results in smooth muscle relaxation
and vasodilation via NO/cGMP pathway [7,8]. Inadequate level
of NO/cGMP leads to ED. Thus, cGMP is the major therapeutically important target to overcome ED by inhibiting the cGMP-specific
phosphodiesterase (PDE-5) enzyme [9]. However, sildenafil a PDE5
inhibitor has been used to treat ED, but it has life-threatening sideeffects
viz. cardiac arrhythmia and hypotension [10] and vascular
or neuronal deficiency like diabetes [11]. Thus, some alternative or
complementary therapeutic approach is the best method to treat
impotence without any side effects. In recent years, a remarkable
outstanding alternative Complementary and Alternative Medicine
(CAM) therapies approach known as Biofield Energy Healing
Treatment (The Trivedi Effect®) have been scientifically reported in
various fields. The human Biofield Energy is a weak electromagnetic
field around of the human body. The Trivedi Effect® can be able to
transform all the living organisms and non-living materials through
a unique energy transmission process [12]
The effects of the CAM therapies have great potential,
which include Johrei, Qi Gong, external qigong, Tai Chi, Reiki,
therapeutic touch, deep breathing, yoga, polarity therapy, pranic
healing, chiropractic/osteopathic manipulation, guided imagery,
meditation, massage, homeopathy, hypnotherapy, special diets,
progressive relaxation, acupressure, acupuncture, relaxation
techniques, mindfulness, Rolfing structural integration, healing
touch, movement therapy, pilates, Ayurvedic medicine, traditional
Chinese herbs and medicines in biological systems both in vitro and
in vivo [12]. Biofield Energy Healing as a CAM showed significant
results in biological studies [13]. Also, the National Center for
Complementary and Alternative Medicine (NCCAM), well-defined
Biofield therapies in the subcategory of Energy Therapies [14].
The Trivedi Effect® has been reported to have created significant
changes in the materials science [15-17], agricultural science
[18,19], microbiology [20-22], biotechnology [23,24], improved
bioavailability [25-27], skin health [27-29], nutraceuticals [30,31],
cancer research [32,33], bone health [34-36], human health and
wellness. Based on the outstanding benefits of The Trivedi Effect®,
the present study was aimed to investigate the effect of Biofield
Treated DMEM on the level of cGMP, in order to eradicate the ED
using standard in vitro assay in Human Endothelial Hybrid Cell Line
(EA. hy926).
Material and Methods
Requirement of Chemicals
Dulbecco’s Modified Eagle’s Medium (DMEM) and fetal
bovine serum (FBS) were obtained from Life Technology, USA.
Antibiotics solution (penicillin-streptomycin) was purchased from
HiMedia, India, while ethylenediaminetetraacetic acid (EDTA) was
purchased from Sigma, USA. Sildenafil citrate was purchased from
Clearsynth, India. All the other chemicals used in this experiment
were analytical grade procured from India.
Cell Culture
Human Endothelial Hybrid Cell Line (EA. hy926) was used as a
test system in this experiment. The cells were maintained in DMEM
growth medium for routine culture supplemented with 10% FBS.
Growth conditions were maintained at 37°C, 5%CO2, and 95%
humidity and subcultured by trypsinization followed by splitting
the cell suspension into new flasks and supplementing with fresh
cell growth medium. Three days before the start of the experiment,
the growth medium of near-confluent cells was replaced with fresh
phenol-free DMEM, supplemented with 10% charcoal-dextran
stripped FBS (CD-FBS) and 1% penicillin-streptomycin [37].
Study Design
The experimental groups consisted of group 1 (G-I) with serumfree
DMEM defined as the untreated DMEM. Group 2 (G-II) consisted
of positive control (sildenafil citrate) at different concentrations.
Further, group 3 (G-III) included DMEM medium (test item group)
with the one-time Biofield Energy Treatment and denoted as BT-I,
while the group 4 (G-IV) included the test item with the two-times
Biofield Energy Treatment and indicated as the BT-II.
Biofield Energy Healing Treatment Strategies
The test item, DMEM was divided into three parts. One part of
the test item was treated with the one-time Biofield Energy Healing
Treatment by a renowned Biofield Energy Healer (The Trivedi
Effect®) and coded as the Biofield Energy Treated DMEM (BT-I),
while the second part was received the two-times Biofield Energy
Healing Treatment and denoted as the BT-II. Further, the third part
did not receive any treatment and defined as the untreated DMEM
group. This Biofield Energy Healing Treatment was provided by
a renowned Biofield Energy Healer, Mahendra Kumar Trivedi,
remotely for~3 minutes. The Biofield Energy Healer was located in
the USA, while the test item was located in the research laboratory
of Dabur Research Foundation, New Delhi, India. This Biofield
Energy Treatment was administered for ~3 minutes through the
Healer’s unique Energy Transmission process remotely to the
test items under the standard laboratory conditions. Mahendra
Kumar Trivedi never visited the laboratory in person, nor had any
contact with the test item (DMEM medium). Further, the untreated
DMEM group was treated with a “sham” healer for comparative
purposes. The “sham” healer did not have any knowledge about the
Biofield Energy Treatment. After that, the Biofield Energy Treated
and untreated samples were kept in similar sealed conditions for
experimental study.
Assessment of PDE-5 Enzyme Inhibition
The cells were counted using an hemocytometer and were
seeded at a density of 0.4 X 106 cells/well in DMEM with 10 % FBS
in 6-well plates. The details test procedure was followed as per
Branton et al. 2018 [38-40]. Increase in cGMP level was determined
as the following equation (1)
% Increase in intracellular cGMP level = {(B-A)/A} x 100-----
-- (1)
Where, B = OD of cells treated with test item and A is the OD of
untreated wells (media treated).
Statistical Analysis
Values were expressed as Mean ± SEM of three independent
experiments. For multiple group comparison, one-way analysis
of variance (ANOVA) was used followed by post-hoc analysis by
Dunnett’s test. Statistically significant values were set at the level
of p≤0.05.
Result and Discussion
Detection of PDE-5 Enzyme Inhibition
The result of the intracellular cGMP level in Ea. hy926 cells
is shown in Figure 1. Sildenafil citrate, used as positive control at
25 μM, 50 μM, and 100 μM exhibited a significant increase in the
intracellular cGMP in Ea. hy926 cells by 34%, 84%, and 234%,
respectively compared to the untreated DMEM group. The onetime
Biofield Energy Treated DMEM (BT-I) group showed 5.03
pmol/mL, while two-times Biofield Energy Treated DMEM (BTII)
group showed 5.58 pmol/mL level of cGMP. Thus, BT-I group
showed a significant (p≤0.001) increase in intracellular cGMP
level by 296.06% in Ea. hy926 cells with respect to untreated
DMEM. Similarly, the BT-II group showed a significant increase in
the intracellular cGMP levels by 339.37% in Ea.hy926 cells than
untreated DMEM. Thus, the data suggest that the two-times Biofield
Energy Healing Treatment (BT-II) showed better results with
respect to the increased cGMP level as compared with the one-time
Biofield Treated DMEM group, which meant that the Biofield Energy
Healing Treatment inhibited PDE-5 enzyme; resulting to the higher
level of cGMP which can help to treat the erectile dysfunction (ED).
Figure 1: Effect of the test items (untreated and Biofield Treated DMEM) on the expression of intracellular cyclic guanosine
monophosphate (cGMP) in human endothelial hybrid (Ea. hy916) cells. BT-I: One-time Biofield Energy Treated DMEM; BT-II:
Two-times Biofield Energy Treated DMEM. ***p≤0.001 vs. untreated DMEM group.
The drugs, which are available in the market for the
treatment of ED, are having serious side-effects along with short
time treatment of ED [41]. Thus, ED can be treated with some
alternative mode of treatment without having any type of adverse
effects. Biofield Energy Healing Treatment is one of the best CAM
approach worldwide to treat various clinical disorders along with
a significant change in different scientific fields. The results are
outstanding and can be comparable with the marketed synthetic
drug, sildenafil citrate. Scientific literature results showed that
relaxation in smooth muscles results in improved level of cGMP
production leading to penile erection [42]. Increase in cGMP results
in decreased level of intracellular calcium, which supports penile
erection [43]. However, cGMP activation is regulated by the PDE-5
enzyme, which is abundant in the corpus cavernosum and results
in improved blood circulation that leads to penile erection [44].
Biofield Energy Healing based DMEM and Biofield Energy Healing
Treatment might work by the relaxation of penile smooth muscles,
which could lead to penile erection.
Conclusion
Erectile dysfunction results an unsatisfactory sex life, mental
stress or anxiety, embarrassment or low self-esteem, relationship
problems, inability to get your partner pregnant, which can
produce lot of pathological implications like hypertension,
hypercholesterolemia, diabetes mellitus, cardiovascular disease,
and depression. Thus, the Biofield Energy Healing Therapy is one of
the best approach to treat various sexual disorders and its related
diseases. The present study results showed that the Biofield Energy
Treated DMEM significantly increased the level of intracellular cGMP
in Ea. hy926 cells compared with the untreated DMEM group. PDE-
5 is the predominant phosphodiesterase, while the intracellular
cGMP level was significantly (p≤0.001) increased by 296.06% in
the one-time Biofield Energy Treated DMEM group (BT-I) in Ea.
hy926 cells compared to the untreated DMEM group. Additionally,
the BT-II group i.e., the two-times blessed in DMEM also showed
a significantly (p≤0.001) increased the level of cGMP by 339.37%
compared with the untreated DMEM group. Henceforth, it can be concluded that the Biofield Energy Treated (The Trivedi Effect®)
DMEM were found to have a significant impact on cGMP level, which
might significantly inhibit the PDE-5 enzyme that leads to the penile
erection. Thus, the Biofield Therapy can be used for the treatment
of numerous sexual disorders viz. hypoactive sexual desire disorder,
fetishistic disorder, dyspareunia, frotteuristic disorder, vaginismus,
exhibitionistic disorder, voyeuristic disorder, sex addiction,
premature or delayed ejaculation (or sexual malfunction or sexual
disorder) improve normal sexual activity, desire, including physical
pleasure, arousal or orgasm, preference, and neurological disorders,
hormonal imbalances, sexual performance, desire disorders (lack of
sexual desire or interest in sex), marital or relationship problems,
effects of a past sexual trauma, feelings of guilt, depression, and
pain disorders (pain during intercourse).
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Tj is the mechanical stress along axis j, νmi is the electro elastic
module, for example, the piezo module, ij sΨ is the elastic compliance
for the control parameter Ψ = const , and the indexes i= 1, 2, … ,
6; j = 1, 2, … , 6; m = 1, 2, 3. The main size along axis i for the electro
elastic actuator is determined us the working length l = {δ , h,b}
in form the thickness, the height or the width for the longitudinal,
transverse or shift piezo effect.
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propagation coefficient, cΨ is the sound speed for the control
parameterΨ = const ,α is the damping coefficient.
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the Laplace transforms of the appropriate displacements and the
forces on the faces 1, 2. For the micro and nano surgical repairs the
structural schemes of the voltage-controlled or current-controlled
piezo actuator are obtained from its mathematical model.
