Antiaging, Cognition and Anti-Inflammatory Potential of the Biofield Energy Treatment in Vitamin D 3 Deficiency Diet (VDD) Induced Sprague Dawley Rats

on this situation authors constructed the current research work to evaluate the impact of Consciousness Energy Healing Treatment on aging after induction of Vitamin D 3 Deficiency Diet (VDD) in Sprague Abstract A proprietary formulation was designed that consist of minerals (zinc, magnesium, iron, calcium, selenium, and copper), vitamins (pyridoxine HCl, cyanocobalamin, ascorbic acid, alpha tocopherol, and cholecalciferol), Panax ginseng extract, β-carotene, and cannabidiol isolate. The present study was aimed to evaluate the impact of Consciousness Energy Healing Treatment (the Trivedi Effect ® ) on a novel test formulation in male Sprague Dawley (SD) rats, fed with vitamin D 3 deficiency G6, G7, G8, and G9 groups, respectively as compared to the G2 group. Altogether, results suggested that the Biofield Treated test formulation and Biofield Energy Treatment per se significantly increased antiaging, cognitive, and anti-inflammatory biomarkers that could be helpful in various aging/psychiatric or inflammatory disorders. Thus, the results showed a significant slowdown of disease progression and all other disease-related complications/symptoms in the preventive Biofield Energy Treatment group and the Biofield Energy Treated Test formulation groups ( viz . G6, G7, G8, and G9) as compared to the disease control group.

Human Biofield Energy has subtle energy that can work effectively [9]. CAM therapies have been practiced worldwide with reported clinical benefits in different health disease profiles [10]. This energy can be harnessed and transmitted by individuals into living and non-living things via the process of Biofield Energy Healing. Biofield Energy Treatment (the Trivedi Effect ® ) has been published in numerous peer-reviewed science journals with significant outcomes in many scientific fields such as cancer research [11,12], microbiology and biotechnology [13][14][15], pharmaceutical science [16][17][18][19], agricultural science [20][21][22], materials science [23][24][25], dietary supplement [26,27], skin health [28,29], human health and wellness. The planned to evaluate the impact of the Biofield Energy Healing Treatment (the Trivedi Effect ® ) on the test formulation for antioxidant action concerning lipid peroxidation, antioxidant activity using standard assays.
Sodium selenate and ascorbic acid were obtained from Alfa Aesar, India. Panax ginseng extract and cannabidiol isolate were obtained from Panacea Phytoextracts, India and Standard Hemp Company, USA, respectively. Other chemicals used in this experiment were analytical grade procured from India.

Experimental animals
Randomly breed male Sprague Dawley (SD) rats with body weight ranges from 200 to 300 gm were used in this study. The animals were purchased from M/s. Vivo Bio Tech, Hyderabad, India.
Animals were randomly divided into nine groups based on their body weights consist of 6 animals of each group. They were kept individually in sterilized polypropylene cages with stainless steel top grill having provision for holding pellet feed and drinking water bottle fitted with stainless steel sipper tube. The animals were maintained as per standard protocol throughout the experiment.

Consciousness energy healing strategies
The test formulation was divided into two parts.

Experimental procedure
Seven days after acclimatization, animals were randomized and grouped based on body weight. All the animals except G1 were fed with Vitamin D3 deficient diet (VDD) from day -12 to till the end of  The Klotho protein expression was determined using Rat Klotho ELISA Kit in rat's CSF in according to the manufacturer's instructions [30].

Assessment of Serum C-reactive protein (CRP)
Serum C-reactive protein were estimated using standard ELISA assay followed by manufacturer instructions. Serum was collected from all the animals after completion of the experiment was examined for level of CRP. The detailed test procedure of the identification of serum C-reactive protein were performed using manufactured instructions as per individual ELISA kit. The CRP level was tested using CUSABIO, ELISA Assay Kit as per manufacturer instructions.

Statistical analysis
The data were expressed as mean ± Standard Error of Mean (SEM) and subjected to statistical analysis using Sigma Plot (Version 11.0). For multiple comparison One-way analysis of variance (ANOVA) followed by post-hoc analysis by Dunnett's test and for between two groups comparison Student's t-test was performed.
The p≤0.05 was considered as statistically significant.

Results and Discussion
Estimation of klotho protein in Cerebrospinal fluids (CSF)

Assessment of CSF biomarker -β-endorphin
The level of β-endorphin in the normal control group (G1)  groups, respectively as compared to the untreated test formulation group (G4). β-endorphin is an endogenous opioid neuropeptide and peptide hormone, considered as cognition, pain and inflammatory biomarker Figure 2. It is produced in certain neurons within the central nervous system and peripheral nervous system to relieve pain when bound to their mu-opioid receptors [33].

Estimation of 5-hydroxy tryptamine (Serotonin) in CSF
The level of serotonin or 5-hydroxy tryptamine ( serotonergic neurotransmission also leads to aging process [34]. In this experiment, the Biofield Energy Treated test formulation had significantly improve the level of serotonin, which might reduce aging process Figure 3.    The effect of the novel test formulation on the level of serum c-reactive protein (CRP) is presented in Figure 1. The serum CRP level in the disease control (vitamin D 3 deficiency) group was 895.29 ± 6.02 ng/mL, which was found to be 98.06% higher than that of the The Biofield Energy Healing Treatment also helped to slow down the disease progression and disease-related complications impacting the overall animals' health. These data suggested that Biofield Energy Treatment per se and Biofield Energy Treated Test formulation in combination would be the best treatment strategy to prevent and protect from the occurrence of any type of disease.

Effect of the test formulation on serum CRP level
Therefore, the Biofield Energy Healing Treatment (the Trivedi Effect ® ) per se might be effective in healthy humans when used as a preventive maintenance therapy to sustain good health, to boost