International Journal of Biomedical Science and Engineering 2017; 5(5): 53-62 http://www.sciencepublishinggroup.com/j/ijbse doi: 10.11648/j.ijbse.20170505.12 ISSN: 2376-7227 (Print); ISSN: 2376-7235 (Online) ® Potential Role of the Trivedi Effect - Biofield Energy Healing on Immunomodulatory Response of Herbomineral Formulation in Male Sprague Dawley Rats Mahendra Kumar Trivedi1, Alice Branton1, Dahryn Trivedi1, Gopal Nayak1, Cathryn Dawn Nykvist1, Celine Lavelle1, Daniel Paul Przybylski1, Dianne Heather Vincent1, Dorothy Felger1, Douglas Jay Konersman1, Elizabeth Ann Feeney1, Jay Anthony Prague1, Joanne Lydia Starodub1, Karan Rasdan1, Karen Mie Strassman1, Leonid Soboleff1, Maire Mayne1, Mary M. Keesee1, Padmanabha Narayana Pillai1, Pamela Clarkson Ansley1, Ronald David Schmitz1, Sharyn Marie Sodomora1, Sambhu Charan Mondal2, Snehasis Jana2, * 1Trivedi Global, Inc., Henderson, Nevada, USA 2Trivedi Science Research Laboratory Pvt. Ltd., Bhopal, Madhya Pradesh, India Email address: [email protected] (S. Jana) *Corresponding author To cite this article: Mahendra Kumar Trivedi, Alice Branton, Dahryn Trivedi, Gopal Nayak, Cathryn Dawn Nykvist, Celine Lavelle, Daniel Paul Przybylski, Dianne Heather Vincent, Dorothy Felger, Douglas Jay Konersman, Elizabeth Ann Feeney, Jay Anthony Prague, Joanne Lydia Starodub, Karan Rasdan, Karen Mie Strassman, Leonid Soboleff, Maire Mayne, Mary M. Keesee, Padmanabha Narayana Pillai, Pamela Clarkson Ansley, Ronald David Schmitz, Sharyn Marie Sodomora, Sambhu Charan Mondal, Snehasis Jana. Potential Role of the Trivedi Effect® - Biofield Energy Healing on Immunomodulatory Response of Herbomineral Formulation in Male Sprague Dawley Rats. International Journal of Biomedical Science and Engineering. Vol. 5, No. 5, 2017, pp. 53-62. doi: 10.11648/j.ijbse.20170505.12 Received: October 30, 2017; Accepted: November 10, 2017; Published: December 5, 2017 Abstract: A new proprietary herbomineral formulation was formulated, consisting of herbal root extract ashwagandha and minerals (zinc, magnesium, and selenium). The objective of this study was to evaluate the immunomodulatory effect of the Biofield Energy Healing (The Trivedi Effect®) Treatment on the test herbomineral formulation in male Sprague Dawley rats. The test formulation was divided into two parts; one was represented as control, while the other part was treated with the Biofield Energy Healing Treatment by eighteen renowned Biofield Energy Healers and defined as the Biofield Energy Treated formulation. Besides, one group of animals was also received Biofield Energy Treatment by same Biofield Energy Healers under similar conditions. The effect of the test formulation was monitored by an estimation of humoral immune response, delay type hypersensitivity, hematology, biochemistry, body weight, feed intake, relative organ weight, and histopathology in male Sprague Dawley rats. The primary antibody titre level was significantly increased by 36.36% in the Biofield Energy Treated formulation (G3); while decreased by 15.09% in the untreated test formulation (G4) compared to the disease control group (G2). The paw volume was significantly increased by 75% in the Biofield Energy Treated group per se at day -15 (G6) compared to the disease control. The level of red blood corpuscle (RBC) was significantly increased by 14.45% in the G6 group compared to the G2 group. The platelet count was significantly increased by 10.32% in the G3 group; while it was decreased by 5.71% in the G4 group compared with the G2 group. There was a significant elevation of serum phosphorus by 6.04% in the G3 group compared to the G2 group. The concentration of magnesium in serum was increased by 13.00% in the Biofield Energy Treated formulation (G3) compared to the disease control group (G2). The concentration of uric acid was significantly decreased by 8.05% in the Biofield Energy Treated formulation (G3) compared to the G2 group. Further, the change in body weight, feed consumption, organ to body weight ratio data, and histopathology examination did not suggest any statistical difference, which depicts that the Biofield Energy Treated test formulation was found to be safe. These data suggest that the Biofield Treated test formulation can be used for autoimmune and inflammatory diseases such as Rheumatoid arthritis, Alzheimer’s disease, Atherosclerosis, Dermatitis, Diverticulitis, Diabetes, etc. along with stress prevention and management and anti-aging by improving overall health. International Journal of Biomedical Science and Engineering 2017; 5(5): 53-62 54 Keywords: Biofield Energy Healers, The Trivedi Effect®, Herbomineral Formulation, Immunomodulation, Humoral Immune Response, Histopathology, Stress Management, Anti-aging 1. Introduction In the last few years, there has been exponential growth been known for its significant impact on various cancerous reported in the herbal medicine sector. In developing and cells [12]. According to many scientific studies, Biofield developed countries alike, medicinal plant-derived drugs Energy Healing has been reported to have significant are gaining popularity due to their eco-friendly nature and outcomes that may prove to be a more cost effective less side effects. Many traditional and complementary alternative to other approaches [13]. Complementary and medicines currently in use are derived from medicinal Alternative Medicines (CAM) are now rising as preferred plants, animals, and minerals, which are commonly used for method of treatment, among which Biofield Therapy (or the prevention and treatment of many diseases [1]. Healing Modalities) is one approach that has been reported However, the use of traditional remedies has gained to have several benefits to enhance physical, mental and importance in cases when conventional medicine is emotional human wellness. However, Biofield Energy can ineffective for certain diseases. The traditional medicines exist in different forms such as kinetic, magnetic, potential, are suitable candidates for new therapeutics due to their electrical, and electromagnetic. The human body has the vast chemical diversity and various biological effects [2]. power to produce low intensity electromagnetic signals Withania somnifera (ashwagandha) is an important known as the Biofield. Thus, a human has the ability to medicinal plant that belongs to the family Solanaceae. It is harness energy from the environment and transmit it to any commonly known as Indian ginseng and is used for various living or nonliving object(s) around the globe. The objects treatments as alternative therapy [3]. Withanolides have always receive the energy and respond in a useful way. This been reported as major active constituents that are isolated process is known as Biofield Energy Healing Treatment from the root and leaves of the ashwagandha plant for (The Trivedi Effect®). Based on the literature data, Biofield biological activity [4]. Apart from its important Energy Treatment in terms of a CAM approach was antibacterial activity, several reports have demonstrated its practiced worldwide [14] in addition to herbal medicine. potent immunomodulatory and anti-tumor activity [5]. The National Center of Complementary and Integrative Preclinical and clinical studies report that each of the active Health (NCCIH) has been recognized and accepted Biofield constituents of ashwagandha have shown Energy Healing as a CAM health care approach in addition immunomodulatory effects in various inflammatory to other therapies, medicines and practices such as natural diseases [6], but the mechanisms of anti- products, deep breathing, yoga, Tai Chi, Qi Gong, inflammatory/immunomodulation remained unknown. chiropractic/osteopathic manipulation, meditation, massage, Selenium, zinc, copper, and magnesium, etc. are highly special diets, homeopathy, progressive relaxation, guided recommended trace elements due to their imagery, acupressure, acupuncture, relaxation techniques, immunomodulatory impact [7, 8]. The coordinated hypnotherapy, healing touch, movement therapy, pilates, interactions of these molecules with the immune cells may rolfing structural integration, mindfulness, Ayurvedic produce an appropriate immune response. Recently, we medicine, traditional Chinese herbs and medicines, prepared a new proprietary herbomineral formulation, naturopathy, essential oils, aromatherapy, Reiki, and cranial which consisted of a combination of the ashwagandha root sacral therapy. To this day, Biofield Energy Healing has had extract and minerals (zinc, magnesium, and selenium). significant impact in the transformation of living organisms These ingredients of the test formulation possess significant and nonliving materials including metals, polymers, anti-inflammatory, antioxidant, anti-infective, anti-viral and ceramics, chemicals, and pharmaceutical compounds. Even immune-modulating properties [5, 7, 9, 10], which plays a further, Biofield Energy Healing Treatment (The Trivedi key role in protecting cells from oxidative stress. Based on Effect®) has been published in numerous peer-reviewed the recent literature, it was reported that the herbomineral science journals due to its significant impacts in the science formulation had exhibited the level of phagocytic index and fields of biotechnology [15-16], genetics [17-19], cancer improved antibody titre to suggest a significant [20-21], microbiology [22-26], materials science [27-30], immunomodulatory response. Further, it was reported that and agriculture [31-34]. the immunomodulatory effect was potentiated in the The authors of this study sought to evaluate the impact of presence of minerals [11], which can be useful for immune- Biofield Energy Treatment (The Trivedi Effect®) on the given compromised patients, autoimmune disorders, cancer, anti- herbomineral formulation, which might improve its stress, anti-aging and in reducing the risk of cardiovascular immunomodulatory function in male Sprague Dawley rats diseases on long term basis. In recent years, Biofield with respect to the humoral immune response, hematological, Energy Treatment (The Trivedi Effect®) has been reported biochemical, body weight, feed consumption, relative organ worldwide as an alternative treatment method which has weight, and histopathology parameters. 55 Mahendra Kumar Trivedi et al.: Potential Role of the Trivedi Effect® - Biofield Energy Healing on Immunomodulatory Response of Herbomineral Formulation in Male Sprague Dawley Rats 2. Materials and Methods were returned in the similar sealed condition and kept in recommended storage condition. 2.1. Chemicals and Reagents 2.4. Antigen (Sheep RBC) Pyrogallol and sodium carboxymethyl cellulose were purchased from Sigma Chemical Co. (St. Louis, MO). The fresh sheep blood was collected aseptically from the Ashwagandha (Withania somnifera) root extract powder was jugular vein of a healthy sheep and transferred immediately procured from Sanat Products Ltd., India. Zinc chloride and to the heparinized tube. The collected erythrocytes were magnesium (II) gluconate hydrate were procured from TCI, separated from plasma by centrifugation (400 g, 10 ºC, 10 Japan. Sodium selenate was procured from Alfa Aesar, minutes), washed twice with the normal saline and then U.S.A. Levamisole hydrochloride was procured from Sigma, further diluted in the normal saline and the samples were U.S.A. All other chemicals used were of analytical grade analyzed using Hematology analyzer (Abbott Model-CD- available in India. 3700). Based on the number of erythrocytes, the samples were further diluted (using normal saline) before injecting to 2.2. Laboratory Animals the rats [35]. Sprague Dawley (SD) male rats were purchased from M/s. 2.5. Experimental Procedure Vivo Bio Tech Ltd., Hyderabad, India. All animals were maintained with 12 hours light and dark cycle during the 24 After 5 days of acclimatization, the animals were hours period alongwith a temperature control 22 ± 3°C, randomized and grouped based on the body weight. Normal humidity of 30% to 70%. The standard chow diet was control (G1) was received oral suspension of 0.5% carboxy procured from M/s. Golden feeds, Mehrauli, New Delhi, methyl cellulose-sodium salt via gavage. The disease control India, which was provided reverse osmosis filtered drinking group (G2) was received pyrogallol through intraperitoneal water ad libitum to all the groups of animals. All the animal (i.p.) route at a dose of 100 mg/kg once daily for 7 days. The experimentation procedures were performed with the Guide G3 and G4 animals were received the Biofield Energy for the Care and Use of Laboratory Animals published by the Treated and untreated test formulations, respectively at US National Institutes of Health. The approval of the 1105.005 mg/kg b.wt, per oral (p.o.). The G5 animals were Institutional Animal Ethics Committee was taken prior to received levamisole at a dose of 50 mg/kg p.o. G6 animals animal experiments. were received the Biofield Energy Treatment per se at day - 15. Further, all the animals except normal control group (G1) 2.3. Biofield Energy Treatment Strategies received pyrogallol at a dose of 100 mg/kg through i.p. route once daily from day 1 to day 7. The animals were treated The herbomineral test formulation was divided into two with the Biofield Energy Treated and untreated herbomineral parts. One part of the test formulation was treated with formulations to the G3 and G4 animals respectively, 1 hour Biofield Energy by renowned Biofield Energy Healers (also known as The Trivedi Effect®) and coded as the Biofield before pyrogallol challenge in the morning once daily for 22 days. The treatment was continued to all the tested groups Energy Treated formulation, while the second part of the test (G1 to G6) with 5 mL/kg body weight dose volume. On day formulation did not receive any sort of treatment and was 7th and day 13th, all the animals in the G2-G6 except normal defined as the untreated test formulation. This Biofield control (G1) were challenged with sheep red blood cells Energy Treatment was provided to the test formulation (sRBC) (0.5 X 109/100 gm; i.p.) as the antigenic material to through a group of eighteen Biofield Energy Healers who sensitize them for immunological parameters. On the days participated in this study and performed the Biofield Energy 13th and 20th, blood sample was collected from retro-orbital Treatment remotely. Eleven Biofield Energy Healers were plexus and subjected to hemagglutination test to evaluate the remotely located in the U.S.A., four were remotely located in humoral immune response. On the same days, the animals Canada, one in the UK, one in Russia and one in Ireland. The were challenged with sRBC (0.5 x 109 cells/50 µL/rat) in test herbomineral formulation was located in the research sub-planter region and on 22nd day (48 hours), paw volume laboratory of Dabur Research Foundation, New Delhi, India. was measured to evaluate cellular immune response. The This Biofield Energy Treatment was administered for 5 body weight and food consumption were measured daily minutes through the Healer’s Unique Energy Transmission before treatment. On day 22, the animals were kept under process remotely to the test formulation under the laboratory fasting over night and on day 23; blood was collected again conditions. Besides, one group of animals was also received from retro-orbital plexus from each animal under isoflurane Biofield Energy Treatment by the same Biofield Energy anaesthesia. Whole blood was analysed for haematological Healers under similar conditions. None of the Biofield parameters and serum was analysed for serum biochemistry. Energy Healers in this study visited the laboratory in person, At the end of the study; animals were euthanized by CO nor had any contact with the herbomineral samples. Further, 2 asphyxiation as per in-house approved standard protocol. the control group was treated with a “sham” healer for Different organs of all animals were excised, weighed and comparative purpose. The sham healer did not have any preserved for histopathological analysis. knowledge about the Biofield Energy Treatment. After that, the Biofield Energy Treated and untreated test formulation International Journal of Biomedical Science and Engineering 2017; 5(5): 53-62 56 2.6. Determination of Humoral Immune Response daily in accordance with in-house protocol [39]. Animals found in a moribund condition or enduring signs of severe On day 13th and 20th, blood was withdrawn from the retro- distress was humanely euthanized. Abnormal findings were orbital plexus of all antigenically challenged rats. recorded with the time of onset and disappearance. Approximately 25 µL of serum was serially diluted with 25 µL of phosphate-buffered saline. The sRBC (0.025 x 109 2.11. Measurement of Relative Organ Weight and cells) was added to each of these dilutions and incubated at Histopathology 37°C for one hour. The rank of minimum dilution that At the end of the experiment, rats were dissected and the exhibited hemagglutination was considered as an antibody titre. The level of antibody titre on day 13th of the experiment whole liver, kidneys, heart, spleen, lungs, whole intestine, eyes, brain, testis, prostate, epididymis, and vas deference was considered as the primary humoral immune response and the day 20th of the experiment was considered as the were excised, weighed, and observed various gross pathological lesions. These organs were trimmed off any secondary humoral immune response [36]. adherent tissue and fat, as appropriate and were weighed in 2.7. Determination of Paw Volume (Delayed Type wet condition as soon as possible to avoid drying. The organ Hypersensitivity) to body weight ratio was determined by comparing with the weight of each organ with the final body weight of each rat. The cellular immune response was assayed by the footpad Defined samples were placed in the neutral buffered formalin reaction method. The edema was induced in the right paw of (10%) for histopathological examination as per standard in- rats by injecting sRBC (0.025 x 109 cells) in the sub-plantar house protocol. region. The change in paw volume after 24 hours (on day 21) was assessed on digital plethysmometer (Pan Lab, Spain). 2.12. Statistical Analysis The mean percentage change in paw volume was considered Data were expressed as mean ± standard error of mean as delayed type of hypersensitivity and as an index of cell- (SEM) and were subjected to one-way analysis of variance mediated immunity. The volume of the left hind paw, injected (ANOVA), and the post-hoc analysis was performed using similarly with phosphate-buffered saline, served as control. Dunnett’s test using Sigma Plot (11.0) statistical software. 2.8. Determination of Hematological and Biochemical Statistical significance was considered at p<0.05. Parameters 3. Results and Discussion On day 23rd of the experiment, blood was collected from the retro-orbital plexus using heparinized and non- 3.1. Determination of Humoral Immune Response heparinized capillary tubes after 12 to 16 hours of fasting. The non-heparinized blood was kept as such from which The results of primary and secondary humoral immune serum was collected and further stored for biochemical responses after administration of test formulation in male analysis. The heparinized blood was directly subjected for Sprague Dawley rats are summarized in the Table 1. Primary the estimation of various hematological parameters using (on day 13th) and secondary (on day 20th) mean standard instrument. The various hematological parameters hemagglutination (HA) antibody titre values were increased such as hemoglobin (Hb), red blood corpuscle (RBC), packed in the disease control group (G2) compared to the normal cell volume (PCV), mean corpuscular volume (MCV), mean control (G1). On day 13th, the disease control group (G2) corpuscular hemoglobin (MCH), mean corpuscular showed significant higher titre value in the primary response, hemoglobin concentration (MCHC) and platelets were which was similar to the untreated group (G4), while in the analyzed in the blood samples. Further, the levels of Biofield Energy Treated formulation group (G3) the antibody magnesium, blood urea nitrogen (BUN), creatinine, uric acid, titre level was significantly increased by 36.36% compared to calcium, phosphorus, potassium, sodium, and chloride ion the G2. The primary antibody titre level was decreased by concentration were analyzed using an Hematology analyzer 15.09% in the untreated test formulation group (G4) (Abbott Model-CD-3700) [37]. compared with the G2 group. The response of primary antibody titre in the levamisole group (G5) was decreased by 2.9. Determination of Body Weight and Feed Intake 36.36% compared with the G2. Moreover, the primary HA titre was also decreased by 12.18% in the Biofield Energy Body weight and feed intake of all the animals were Treatment group per se at day -15 (G6) compared to the G2. measured once daily before dosing. Briefly, the weight of On day 20th, the secondary antibody titre level was reduced daily feed supply and the left-over amount by the following by 5.25% in the Biofield Energy Treated group (G3) day was recorded and the difference was taken as the daily compared to the G2; while it was decreased by 15.79% in the feed intake. The average of the feed intake was computed for untreated test formulation (G4) compared to the G2. The every three days of the experimental period [38]. secondary HA titre was decreased by 13.15% in the Biofield 2.10. Clinical Signs and Symptoms Energy Treated group per se at day -15 (G6) compared to the G2. All the animals were observed for the clinical signs once 57 Mahendra Kumar Trivedi et al.: Potential Role of the Trivedi Effect® - Biofield Energy Healing on Immunomodulatory Response of Herbomineral Formulation in Male Sprague Dawley Rats Table 1. The effect of the test formulation on humoral immune response in 3.2. Estimation of Delayed Type Hypersensitivity (Paw male Sprague Dawley rats. Volume) Group Primary HA Titre Secondary HA Titre The effects of the Biofield Energy Treated and untreated G1 1.00 ± 0.00 1.00 ± 0.00 G2 5.50 ± 1.2 25.33 ± 4.34 formulation on delayed type hypersensitivity (DTH) response G3 7.5 ± 2.06 24.00 ± 3.58 are shown in Figure 1. The levamisole group (G5) showed G4 4.67 ± 0.67 21.33 ± 3.37 significant (p<0.01) increase in the paw volume compared to G5 3.50 ± 0.5 24.00 ± 3.58 the disease control (G2) group. The paw volume was G6 4.83 ± 1.11 22.00 ± 4.82 significantly increased by 75% in the Biofield Energy HA: Hemagglutination, All the values are expressed as the mean ± SEM; Treatment group per se at day -15 (G6) compared to the The primary responses of mean HA titre was recorded on day 13th and disease control (G2). secondary response on day 20th of the experimental period. G1: Normal control; G2: Disease control (pyrogallol); G3: Biofield Energy Treated test formulation; G4: Untreated test formulation; G5: Reference item (levamisole); G6: Biofield Energy Treatment group per se at day -15. Figure 1. Effect of the test formulation on rat paw volume (delayed-type hypersensitivity). G1: Normal control; G2: Disease control; G3: Biofield Energy Treated test formulation; G4: Untreated test formulation; G5: Reference item (levamisole); G6: Biofield Energy Treatment group per se at day -15. The values are represented as mean ± SEM (n=6). **p≤0.01 vs disease control. 3.3. Determination of Hematological Parameters levamisole (G5) and Biofield Energy Treatment group per se at day -15 (G6), respectively compared to the disease control The effect of the Biofield Energy Treated formulation on (G2). Literature reported that ashwagandha prevented various hematological parameters is shown in the Table 2. myelosuppression and increase the platelet count and body The level of red blood corpuscle (RBC) was reduced by weight [40, 41]. The level of packed cell volume (PCV) was 4.52% in the disease control group (G2) compared to the significantly increased by 21.61%, 17.29%, and 28.32% in normal control group (G1). Besides, RBC was significantly the untreated test formulation (G4), levamisole (G5), and increased by 13.03%, 12.91%, and 14.45%, in the untreated Biofield Energy Treatment group per se at day -15 (G6), test formulation group (G4), levamisole (G5), and Biofield respectively compared to the disease control (G2). However, Energy Treated group per se at day -15 (G6), respectively; the level of MCH was decreased by 3.97%, 9.00%, 15.25%, while the level of RBC was decreased in the Biofield Energy and 15.94% in the Biofield Energy Treated test formulation Treated formulation (G3) compared to the disease control (G3), untreated test formulation group (G4), levamisole (G5), group (G2). The Biofield Energy Treatment group per se at and Biofield Energy Treatment group per se at day -15 (G6), day -15 (G6) showed better improvement in the formation of respectively compared to the disease control (G2). RBC compared to the untreated test formulation group (G4). The level of MCHC was significantly reduced by 9.30%, Hemoglobin (Hb) was increased by 9.48% in the untreated 13.85%, and 14.56% in the untreated test formulation (G4), test formulation group (G4); while reduced by 5.34% in the levamisole (G5), and Biofield Energy Treatment group per se Biofield Energy Treated test formulation (G3) compared to at day -15 (G6), respectively compared to the disease control the G2. Moreover, the level of Hb was significantly increased (G2). The value of RDW-CV was increased by 8.33%, by 9.74% in the Biofield Energy Treatment group per se at 25.00%, 41.67%, and 33.33% in the Biofield Energy Treated day -15 (G6) compared to the disease control group (G2). formulation (G3), untreated test formulation (G4), levamisole The platelet count was significantly increased by 10.32% group (G5), and Biofield Energy Treatment group per se at in the Biofield Energy Treated group (G3); while platelet day -15 (G6), respectively compared to the disease control count was decreased by 5.71% in the untreated test (G2). It was reported that W. somnifera extract was non-toxic formulation (G4) compared to the G2. Besides, the reduction to human erythrocytes at different concentrations [42]. The of platelets was observed by 6.61% and 30.91% in the present experimental results showed the Biofield Energy International Journal of Biomedical Science and Engineering 2017; 5(5): 53-62 58 Treated test formulation did not show any toxic effect on disease control. Besides, the minerals present in the RBC, as no significant change was observed in different herbomineral formulation were reported to be safe and good treatment groups with respect to the both normal control and therapeutic effect [43]. Table 2. Evaluation of hematology parameters after treatment with the test formulation in male Sprague Dawley rats. RBC Platelet Count Group Hb (gm/dL) PCV (%) MCV (fl) MCH (pg) MCHC (%) RDW-CV (106/(cid:1) L) (thousand/mm3) G1 8.84 ± 0.22 16.40 ± 0.37 49.12 ± 1.10 55.60 ± 0.63 18.55 ± 0.18 33.38 ± 0.14 1120.00 ± 83.03 0.12 ± 0.00 G2 8.44 ± 0.12 15.92 ± 0.12 47.60 ± 0.37 56.45 ± 0.77 18.88 ± 0.33 33.43 ± 0.20 961.00 ± 87.64 0.12 ± 0.00 G3 8.33 ± 0.29 15.07 ± 0.39 45.00 ± 1.18 54.17 ± 1.40 18.13 ± 0.51 33.48 ± 0.17 1060.17 ± 55.50 0.13 ± 0.00 G4 9.54 ± 0.55 17.43 ± 0.56 57.88 ± 3.54* 56.72 ± 0.52 17.18 ± 0.54 30.32 ± 1.02 906.17 ± 90.17 0.15 ± 0.01 G5 9.53 ± 0.28 16.10 ± 0.22 55.83 ± 0.88* 55.70 ± 0.47 16.00 ± 0.10 28.80 ± 0.19 897.50 ± 96.83 0.17 ± 0.02 G6 9.66 ± 0.46 17.47 ± 0.21 61.08 ± 0.66* 55.40 ± 0.61 15.87 ± 0.16** 28.55 ± 0.15* 664.00 ± 71.14* 0.16 ± 0.00 G1: Normal control; G2: Disease control; G3: Biofield Energy Treated test formulation; G4: Untreated test formulation; G5: Reference item (levamisole); G6: Biofield Energy Treatment group per se at day -15. The values are represented as mean ± SEM (n=6). *p≤0.05 and **p≤0.01 compared to the disease control. Hb: Hemoglobin; RBC: Red blood count; PCV: Packed cell volume; MCV: Mean corpuscular volume; MCH: Mean corpuscular hemoglobin; MCHC: Mean corpuscular hemoglobin concentration; RDW-CV: Red cell distribution width and volume. 3.4. Effect of the Biofield Energy Treated Test Formulation (G5), respectively compared to the disease control (G2). In on Serum Biochemistry addition to, the serum concentration of BUN was reduced by 16.03% in the Biofield Energy Treatment group per se at day - The effect of the Biofield Energy Treated test formulation 15 (G6); while in the others tested groups it was increased on hematological parameters is shown in the Table 3. Among compared to the disease control (G2). Moreover, the serum the ions estimated, there was a significant elevation of concentration of uric acid (UA) was significantly decreased by phosphorus by 6.04%, 20.11%, and 3.63% in the Biofield 8.05%, 26.83%, and 53.66% in the Biofield Energy Treated Energy Treated formulation (G3), untreated test formulation test formulation (G3), levamisole (G5), and Biofield Energy (G4), and levamisole (G5), respectively compared to the Treatment group per se at day -15 (G6), respectively compared disease control (G2). Similarly, the level of potassium was with the G2 group. Here, the Biofield Energy Treated increased by 1.70%, 15.15%, and 2.27% in the Biofield herbomineral formulation showed the better effect by reducing Energy Treated test formulation (G3), untreated formulation the UA than the untreated test formulation. These results might (G4), and levamisole (G5), respectively; while it was reduced be due to the positive effect of the Biofield Energy Healing to by 11.36% in the Biofield Energy Treatment group per se at the novel herbomineral formulation, which could be very day -15 (G6) compared to the disease control (G2). The helpful in immunocompromised patients. Besides, the levels of concentration of magnesium was increased by 13.00%, calcium, creatinine, sodium and chloride ions were altered in 30.67%, and 6.00% in the Biofield Energy Treated formulation all the treated groups compared with the disease control. (G3), untreated test formulation (G4), and levamisole group Table 3. Estimation of biochemical parameters after treatment with the test formulation in male rats. Magnesium Blood Urea Creatinine Uric Acid Calcium Phosphorus K+ Group Na+ (Meq/L) Cl- (mEq/L) (mg/dL) (mg/dL) (mg/dL) (mg/dL) (mg/dL) (mg/dL) (mEq/L) G1 3.01 ± 0.14 41.30 ± 0.66 0.52 ± 0.02 3.60 ± 0.25 10.68 ± 0.57 9.28 ± 0.21 150.67 ± 0.21 5.32 ± 0.11 102.83 ± 0.48 G2 3.00 ± 0.24 50.77 ± 2.96 0.48 ± 0.04 4.10 ± 0.77 10.38 ± 0.22 9.10 ± 0.50 151.00 ± 0.68 5.28 ± 0.29 102.83 ± 1.05 G3 3.39 ± 0.13 60.48 ± 4.84 0.53 ± 0.03 3.77 ± 0.70 10.35 ± 0.12 9.65 ± 0.34 152.83 ± 0.98 5.37 ± 0.16 103.00 ± 0.68 G4 3.92 ± 0.13 55.80 ± 3.62 0.47 ± 0.03 4.32 ± 0.48 10.63 ± 0.26 10.93 ± 0.43 150.07 ± 0.67 6.08 ± 0.38 101.33 ± 0.71 G5 3.18 ± 0.08 55.68 ± 3.00 0.50 ± 0.04 3.00 ± 0.52 10.13 ± 0.18 9.43 ± 0.14 150.83 ± 0.75 5.40 ± 0.16 102.67 ± 0.61 G6 2.95 ± 0.15 42.63 ± 3.35 0.58 ± 0.03 1.90 ± 0.46 10.45 ± 0.13 8.73 ± 0.25 153.33 ± 0.76 4.68 ± 0.07 104.17 ± 0.54 All values are presented as mean ± SEM (n=6). G represents as group; G1: Normal control; G2: Disease control; G3: Biofield Energy Treated test formulation; G4: Untreated test formulation; G5: Reference item (levamisole); G6: Biofield Energy Treatment group per se at day -15. 3.5. Effect of the Test Formulation on Body Weight, Feed showed slight reduction of feed consumption compared to the Intake, and Organ to Body Weight Ratio disease control, which might be due to physiological variation in male rats. It is assumed that the Biofield Treated formulation The effect of the Biofield Energy Treated test formulation was safe and effective with respect to both consumption of administration on animal weight parameters in male rats was feed and consequently change of body weight. The results analyzed and presented in Table 4. The results reflect the suggest that no significant change throughout the experimental change in body weight, as final weights were increased among period in relative organ weight parameters like liver, lungs, all the tested groups. The mean body weight percentage kidneys, brain, heart, eyes, spleen, whole intestine, testis, difference in the Biofield Energy Treated test formulation prostate, epididymis, and vas deference compared to the group and untreated test formulation group did not have any normal and disease control groups (Table 4). The result of significant difference compared with the disease control group. relative organ weight was slightly increased in the disease The Biofield Energy Treated test formulation group (G3) control group; while after treatment with the Biofield Energy 59 Mahendra Kumar Trivedi et al.: Potential Role of the Trivedi Effect® - Biofield Energy Healing on Immunomodulatory Response of Herbomineral Formulation in Male Sprague Dawley Rats Treated test formulation, the organ weight reached to normal level similar to the control group. Table 4. Effect of the test formulation on organ weight parameters in male Sprague Dawley rats. Relative organ weight (%) G1 G2 G3 G4 G5 G6 Liver 3.60 ± 0.17 3.99 ± 0.19 3.56 ± 0.22 3.55 ± 0.09 3.92 ± 0.13 3.45 ± 0.12 Lungs 0.62 ± 0.03 0.71 ± 0.04 0.61 ± 0.04 0.61 ± 0.04 0.60 ± 0.02 0.54 ± 0.03 Kidneys 0.84 ± 0.02 0.91 ± 0.02 0.85 ± 0.06 0.80 ± 0.02 0.96 ± 0.03 0.83 ± 0.02 Brain 0.60 ± 0.01 0.59 ± 0.02 0.55 ± 0.02 0.58 ± 0.02 0.63 ± 0.03 0.60 ± 0.02 Heart 0.44 ± 0.03 0.40 ± 0.02 0.35 ± 0.01 0.36 ±0.01 0.37 ± 0.02 0.37 ± 0.01 Eyes 0.08 ± 0.00 0.08 ± 0.02 0.09 ± 0.01 0.08 ± 0.01 0.09 ± 0.01 0.09 ± 0.01 Spleen 0.21 ± 0.01 0.29 ± 0.03 0.22 ± 0.01 0.20 ± 0.03 0.27 ± 0.02 0.28 ± 0.02 Whole intestine 5.47 ± 0.09 5.00 ± 0.24 6.64 ± 0.28 7.25 ± 0.40 6.44 ± 0.58 4.86 ± 0.22 Testis 0.96 ± 0.07 0.92 ± 0.04 0.92 ± 0.07 0.96 ± 0.05 1.01 ±0.03 0.91 ± 0.10 Prostrate 0.27 ± 0.02 0.29 ± 0.03 0.22 ± 0.01 0.26 ± 0.03 0.25 ± 0.01 0.26 ±0.02 Epididymis 0.38 ± 0.02 0.37 ± 0.03 0.36 ± 0.03 0.36 ± 0.04 0.42 ± 0.03 0.38 ± 0.03 Vas deference 0.07 ± 0.01 0.07 ± 0.00 0.07 ± 0.00 0.07 ± 0.00 0.07 ± 0.01 0.08 ± 0.01 Values are presented as mean ± SEM (n=6). G represents as group; G1: Normal control; G2: Disease control; G3: Biofield Energy Treated test formulation; G4: Untreated test formulation; G5: Reference item (levamisole); G6: Biofield Energy Treatment group per se at day -15. The organ to body weight ratio is a useful index for the 3.6. Histopathological Study identification of swelling, atrophy or hypertrophy [44]. The increase in body weight or organ weight with the exposure of The effect of the Biofield Energy Treated test formulation any compound in the animals during experiment suggest the on histopathological findings in male rats is shown in the hypertrophy, while decrease in the relative weight indicated Figure 2. No significant differences were observed either in the atrophy. The increase in body weight and organ-body gross or microscopic observation of the tested organs. ratio might be correlated with the sign of product toxicity, but Histopathological study results also suggest that no treatment- the experimental results suggest that there was not much related histopathological findings were reported in all the change in most of the vital organs, which depicts that the test experimental animals compared with the control group. The formulation was non-toxic to the animals throughout the detailed histopathological images of microscopic sections of exposure period at a dose of 1105.005 mg/kg. the organs are presented in Figure 2. Mild vacuolization in centrizonal hepatocytes was observed in few animals in the untreated group (G4). All other organs of animals were devoid of any microscopically changes (Figure 2). Figure 2. Histopathological photomicrograph of major organs tested after Biofield Energy Treated test formulation in male Sprague Dawley rats. All the tissues were sectioned transversely and stained with hematoxylin and eosin. G1: Normal control; G2: Disease control; G3: Biofield Energy Treated test formulation; G4: Untreated test formulation; G5: Reference item (levamisole); G6: Biofield Energy Treatment group per se at day -15. International Journal of Biomedical Science and Engineering 2017; 5(5): 53-62 60 Biofield Energy Healing has been reported to be effective Arthritis, Sjogren Syndrome, Systemic Lupus Erythematosus, in cancer treatment by reducing the level of cytokines [45- Type 1 Diabetes, Alopecia Areata, Crohn’s Disease, 47]. Besides, The National Center for Fibromyalgia, Vitiligo, Psoriasis, Scleroderma, Chronic Complementary/Alternative Medicine (NCCAM), 34% of Fatigue Syndrome and Vasculitis, as well as inflammatory adults in U.S. populations depends on some forms of disorders such as Asthma, Ulcerative Colitis, Alzheimer’s complementary health approach, among which energy Disease, Atherosclerosis, Dermatitis, Diverticulitis, Hepatitis, medicine is one of them. Complementary and Alternative Irritable Bowel Syndrome, Parkinson’s Disease and stress etc. Medicine (CAM) has several advantages instead of the Further, the Biofield Energy Healing Treated test formulation current preferred treatment approach [48]. The new can also be used in the prevention of immune-mediated tissue proprietary herbomineral formulation that might act as better damage in cases of organ transplants (for example heart immunomodulatory medicine in the future due to its anti- transplants, kidney transplants and liver transplants), for anti- inflammatory and immunomodulatory effects. Therefore, it is aging, stress prevention and management, and in the assumed that the Biofield Energy Treated herbomineral improvement of overall health and quality of life. formulation might be considered as a safe dietary supplement for boosting the immuno response. Acknowledgements 4. Conclusions The authors are grateful to Dabur Research Foundation, Trivedi Science, Trivedi Global, Inc., and Trivedi Master The primary antibody titre level was significantly increased Wellness for their support throughout the work. by 36.36% in the Biofield Energy Treated test formulation group (G3) compared to the disease control group and also showed better result than the untreated test formulation (G4). References The paw volume was significantly increased by 75% in the Biofield Energy Treatment group per se at day -15 (G6) [1] Rishton GM (2008) Natural products as a robust source of new drugs and drug leads: Past successes and present day compared to the disease control. The platelet count was issues. Am J Cardiol 101: 43D-9D. significantly increased by 10.32% in the G3 group compared to the disease control group and also showed better response [2] Mukhtar M, Arshad M, Ahmad M, Pomerantz R, Wigdahl B, than the G4 group. 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