The Role of Biofield Energy Treated DMEM in Erectile Dysfunction using Detection of cGMP Levels in Human Endothelial Cell Line

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.

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]. 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][16][17], agricultural science [18,19], microbiology [20][21][22], biotechnology [23,24] [32,33], bone health [34][35][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). 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%CO 2 , 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.

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

Assessment of PDE-5 Enzyme Inhibition
The cells were counted using an hemocytometer and were seeded at a density of 0.4 X 10 6 cells/well in DMEM with 10 % FBS in 6-well plates. The details test procedure was followed as per Branton et al. 2018 [38][39][40]. Increase in cGMP level was determined

Detection of PDE-5 Enzyme Inhibition
The result of the intracellular cGMP level in Ea. hy926 cells is shown in Figure 1.  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