Effects of distant biofield energy healing on adults associated with psychological and mental health-related symptoms: a randomized, placebo-controlled, double-blind study.

Background: In recent years, popularity of Biofield energy healing therapy has increased tremendously parallel to conventional therapy. However, due to insufficient scientific studies on biofield therapies, authors planned this clinical trial to examine the safety and effectiveness of biofield therapy in treating symptoms of psychological and mental health and to explore parameters for its effectiveness. Methods: One hundred fourteen participants (55 male, 59 female) underwent clinical trials. This trial was randomized, placebo-controlled, three parallel-groups, double-blind, and single-center with subjects who have one or more psychological and mental symptoms. Two sessions of distant (virtual) biofield energy attunement were given on days 0 and 90 for about 5 minutes to the subjects of biofield intervention group (n=39). Besides, control (n=35) and sham control (n=40) group subjects also received kind of treatment in a similar manner to nullify baseline responses. Subjects were assessed by psychological questionnaire scoring using standard scale of assessment and safety parameters. Results: Perceived psychological symptoms/scores (fatigue/tiredness, sleep disturbances, stress, cognitive impairment, loss of memory, mental restlessness, emotional trauma, anxiety, depression, confusion, financial crises and dissatisfaction, low libido, motivation, confidence, lack of perception, relationship, and social behaviors, etc.) were significantly (p <.0001) improved in biofield intervention group compared to the naïve control and sham control groups. Besides, biofield intervention did not show any study-related adverse effects in all three groups throughout the trial. Conclusions: The distant (virtual) biofield energy healing therapy significantly improved psychological and mental health-related symptoms without affecting safety concerns, and improved overall health and quality of life.

Assessment of cognitive-motor functions in adults with perceived neuropsychological problems using NIH toolbox after remote biofield energy treatment as non-pharmacological intervention: A randomized double-blind placebo controlled trial.

Non-pharmacological interventions include physical activity, biofield energy therapy, reiki, Tai chi, and therapeutic touch. However, no reports analyzed the effectiveness of biofield therapy on cognition and motor function performance in adult subjects. The study aimed to investigate the impact of remote biofield energy healing therapy on cognition and motor functioning in adults with self-perceived neuropsychological impairments. This was a randomized double-blind clinical trial that involved 114 participants with self-perceived neuropsychological impairments. The participants were divided into three groups (control, sham control, and biofield intervention). Cognitive and motor function scores were assessed using the NIH Toolbox at baseline (day 0), day 90, and day 180. The biofield treatment group showed significant improvements in language function (p < 0.0001), working memory (p < 0.0001), and episodic memory (p < 0.0001) scores. Other cognitive functions also improved, although not statistically significant. The biofield intervention group also demonstrated significant enhancements (p < 0.05 to p < 0.0001) in locomotion, standing balance, dexterity, grip strength, and muscle endurance. No adverse effects were reported. The results suggest that remote biofield energy therapy is a safe, noninvasive intervention that improves cognitive and motor functions in adults. Further research is needed to understand its clinical benefits.

The Improvement of Cognitive-Motor Function Scores Using the NIH-Toolbox in Adults Subjects After Biofield Energy Therapy: A Randomized Controlled Double-blind Trial.

Background: There is a growing interest for the use of an alternative treatment approach like biofield energy therapy for managing various diseases. However, the effectiveness of distance mode of biofield energy therapy on neuropsychological function has not been examined so far. Primary Study Objective: The study was aimed to evaluate the effect of biofield energy therapy via distant mode for the benefits of neuropsychological functions in adult subjects. Methods: It comprised of three parallel groups (control, sham control, and biofield intervention). The cognitive-motor functioning scores were monitored using NIH-Toolbox in 119 adult subjects on day 0, day 90, and day 180. Setting: Double-blind, single-center, randomized controlled trial (RCT). Participants: Adult human subjects (male and female) with one or more psychological symptoms. Intervention: Biofield energy therapy through distant/remote (virtual) mode. Primary outcome measures: The cognitive-motor functioning scores. Results: The cognitive-motor functioning scores were significantly improved in the biofield intervention group compared to the control and sham control groups: cognition- language (P < .0057 to P < .0001), working memory (P < .0001), episodic memory function (P < .0369 to P < .0001), and executive function (P < .0386 to P < .0029); motor- locomotion P < .0001), standing balance (P = .0144 to P = .0019), dexterity (P = .0474 to P < .0001), grip strength (P = .0132 to P = .0078) and endurance (P = .0041 to P = .0004). No adverse effects were observed throughout the study. Physical vital signs and blood parameters were within the normal ranges during the entire study period. Conclusions: Overall, cognitive and motor functions significantly improved in the blessing intervention group compared to the naïve control and sham control groups, which ultimately improved the neuropsychological function and quality of life in adult subjects.

The use of biofield energy therapy as complementary and alternative medicine in human health care system: a narrative review and potential mechanisms.

INTRODUCTION: There has been accumulating interest in the application of biofield therapy as complementary and alternative medicine (CAM) to treat various diseases. The practices include reiki, qigong, blessing, prayer, distant healing, known as biofield therapies. This paper aims to state scientific knowledge on preclinical and clinical studies to validate its potential use as an alternative medicine in the clinic. It also provides a more in-depth context for understanding the potential role of quantum entanglement in the effect of biofield energy therapy. CONTENT: A comprehensive literature search was performed using the different databases (PubMed, Scopus, Medline, etc.). The published English articles relevant to the scope of this review were considered. The review gathered 45 papers that were considered suitable for the purpose. Based on the results of these papers, it was concluded that biofield energy therapy was effective in treating different disease symptoms in preclinical and clinical studies. SUMMARY: Biofield therapies offer therapeutic benefits for different human health disorders, and can be used as alternative medicine in clinics for the medically pluralistic world due to the growing interest in CAM worldwide. OUTLOOK: The effects of the biofield energy therapies are observed due to the healer's quantum thinking, and transmission of the quantum energy to the subject leads to the healing that occurs spiritually through instantaneous communication at the quantum level via quantum entanglement.

Vitamin D3 supplementation improves spatial memory, muscle function, pain score, and modulates different functional physiological biomarkers in vitamin D3 deficiency diet (VDD)-induced rats model.

BACKGROUND: Vitamin D Deficiency is recognized as a pandemic, which is associated with high mortality. An inadequate level of vitamin D is associated with autoimmune diseases, hypertension, and cancer. The study was aimed to assess the pharmacological effects of chronic vitamin D3 supplementation on the manipulation diet regiment of deprived cholecalciferol (vitamin D3 deficient diet, VDD) rats. METHODS: Memory performance (Y-maze task), muscular function (muscle grip strength), and pain score (pressure application measurement meter) were measured. Functional biomarkers were measured using ELISA method in different matrix viz. in serum (parathyroid hormone; PTH, calcitonin, thyroxine, and C-reactive protein; CRP, 25-OH Vit D3), and in CSF (klotho and ß-endorphin). 25-OH Vit D3 was also estimated in liver and kidney homogenate using ELISA. Vitamin D receptor (VDR) was measured spectrophotometrically in liver and adipose tissue. RESULTS: VDD-induced rats showed a decrease in number of entries and time spent in the novel arm and spontaneous alternations in the Y-maze task. Significant improvements of neuromuscular function and pain score after addition of vitamin D3. In comparison to the VDD group, VDR expression (liver) and active metabolites of vitamin D3 (25-OH vit.D3) in serum were significantly higher by 48.23% and 280%, respectively. The PTH and CRP levels were significantly reduced by 32.5% and 35.27%, respectively, whereas calcitonin was increased by 36.67% compared with the VDD group. Klotho and ß-endorphin expressions in cerebrospinal fluid were significantly elevated by 19.67% and 133.59%, respectively, compared to VDD group. CONCLUSIONS: Overall, the results indicate that supplementation of cholecalciferol significantly improved spatial memory impairment, VDR expression, and may provide an opportunity to manage vitamin D deficiencies.

The role of biofield energy treatment on psychological symptoms, mental health disorders, and stress-related quality of life in adult subjects: A randomized controlled clinical trial.

Background: Western and Eastern cultures have practiced biofield energy therapy for thousands of years, but empirical research on effectiveness of this therapy is relatively nascent. Study was aimed to assess the safety and efficacy of biofield therapy on psychological symptoms and mental health disorders in adult subjects. Methods: Seventy-seven participants (39 male and 38 female) underwent clinical trial. This trial was simple randomized, placebo-controlled, parallel-group, open-label, and single-center with subjects, who have one or more psychological symptoms. Two sessions of biofield energy attunement were given in-person at day 0 and 90 for 3 min (treatment group, n = 35) and others allocated to naive attunement (placebo group, n = 42). Subjects were assessed psychological questionnaire scoring using standard scale of assessment and levels of physiological biomarkers in serum were determined by parameter-specific ELISA. Results: Perceived psychological symptoms/scores (asthenia, sleep disturbances, anxiety, depression, stress, confusion, future fearness, sexual desireness, motivation, confidence, emotional trauma, etc.) were significantly (p ≤ 0.0001) improved in the treatment group than placebo control group. Furthermore, physiological biomarkers: vitamins (B12, C, and D3 metabolites), immune biomarker (CD8+CD28-), neurotransmitters (acetylcholine, noradrenaline, and dopamine), hormones (oxytocin, 17-ß-estradiol, and insulin), and antiaging protein (klotho) levels were significantly (p ≤ 0.001) increased in treatment group than placebo. Proinflammatory cytokines (TNF-α, IL-1ß, IL-6, and IL-8) and oxidative stress biomarkers (isoprostane and oxidized LDL) were reduced in treatment group compared with placebo. Conclusions: Results suggest that experimenter's biofield energy plays a significant role in information transfer processes that contribute to individual's state of physical, mental, emotional, and spiritual well-being as well as improved overall health and quality of life.

Simultaneous identification and quantification of pentacyclic triterpenoids and phenolic compounds from the leaves of Boswellia serrata using LC-MS/MS tandem mass spectrometry.

Boswellia serrata (B. serrata) is an important medicinal plant widely used as dietary supplements to provide a support for osteoarthritic and inflammatory diseases. The occurrence of triterpenes in leaves of B. serrata is very little or none. Therefore, the qualitative and quantitative determination of phytoconstituents (triterpenes and phenolics) present in the leaves of B. serrata is very much needed. The aim of this study was to develop an easy, rapid, efficient and simultaneous liquid chromatography-mass spectrometry (LC-MS/MS) method for the identification and quantification of the compounds present in the leaves extract of B. serrata. The purification of ethyl acetate extracts of B. serrata was performed by solid phase extraction method, followed by HPLC-ESI-MS/MS analysis. Chromatographic parameters of the analytical method included negative electrospray ionization (ESI-) with a flow of 0.5 mL/min in gradient mode consisting of acetonitrile (A) and water (B) containing 0.1% formic acid, at 20 °C. Total 19 compounds (13 triterpenes and 6 phenolic compounds) were separated, and simultaneously quantified using a validated LC-MS/MS method with high accuracy and sensitivity. Good linearity was obtained with r2 > 0.973 in the calibration range. The overall recoveries were in a range between 95.78 and 100.2% with relative standard deviations (RSD) below 5% for the entire procedure of matrix spiking experiments. Overall, there was no ion suppression from the matrix. The quantification data showed that the total amount of triterpenes and phenolic compounds in the leaves of B. serrata ethyl acetate extract samples ranged from 14.54 to 102.14 mg/g and 2.14 to 93.12 mg/g of dry extract, respectively. This work provides, for the first time, a chromatographic fingerprinting analysis on the leaves of B. serrata. A rapid, efficient, and simultaneous liquid chromatography-mass spectrometry (LC-MS/MS) method was developed and used for the both identification and quantification of triterpenes and phenolic compounds in the leaves extracts of B. serrata. The method established in this work can be used as quality-control method for other market formulations or dietary supplements containing leaf extract of B. serrata.

Effects of Cannabidiol Interactions with CYP2R1, CYP27B1, CYP24A1, and Vitamin D3 Receptors on Spatial Memory, Pain, Inflammation, and Aging in Vitamin D3 Deficiency Diet-Induced Rats.

Introduction: The study was planned to investigate memory-enhancing, anti-inflammatory, and antiaging potential of cannabidiol (CBD) on vitamin D3 deficient diet (VDD)-induced rats. Materials and Methods: Cytochrome P-450 enzymes were analyzed by RT-PCR method and others biomarkers by enzyme-linked immunosorbent assay. Results: CYP2R1 and CYP27B1-mRNA were significantly increased by 39.29 and 38.37%, respectively, while; CYP24A1-mRNA was significantly reduced by 21.39% compared to VDD. Vitamin D3 receptor protein expression was significantly increased by 148.3%, 60.48%, and 142.03% in liver, kidney, and brain, respectively, compared to VDD group. Vitamin D3 metabolites and serotonin were significantly increased more than 60% and 100%, respectively, compared to VDD. Spatial memory (in terms of total distance, escape latency) and pain score were improved compared to VDD. Cytokines were significantly reduced than VDD. Besides, levels of superoxide dismutase (49.61%), glutathione peroxidase (178.87%), acetylcholine (25.40%), and klotho (145.57%) were significantly increased than VDD. Conclusions: Study findings supported that CBD interacts with CYP2R1, CYP27B1, CYP24A1, and vitamin D receptors, resulting in increased vitamin D3 metabolites, which improved memory, pain tolerance, reduced inflammation, and aging through modulating antioxidative enzymes, cytokines, and neurotransmitters in VDD-induced rats.

Evaluate the Safety and Efficacy of a Biofield Energy Treated Proprietary Dietary Supplement (TRI 360™) on Psychological Symptoms, Mental Disorders, Emotional Well-being, and Quality of Life in Adult Subjects.

Background: The demand for "energy healing" is growing rapidly in the United States and other developed countries. To date, scanty clinical trials have been conducted to evaluate its clinical safety, efficacy, and cost-effectiveness. Primary Study Objective: The study aimed to investigate the safety and efficacy of the Blessed TRI 360TM capsule (Biofield Energy Treated Proprietary Dietary Supplement Capsules Powered by the Trivedi Effect®). Methods/Design: A total of 77 adult subjects (male and female) aged 20-45 years with one or more psychological symptoms were enrolled in the trial. The subjects were randomized into two groups: placebo control and treatment group. The treatment group (n = 35) received the Blessed TRI 360TM capsule twice a day, once in the morning and once in the evening. The placebo control group (n = 42) did not receive any treatment/capsule. Setting: An open-label, single-center, randomized controlled trial (RCT). Participants: Adult human subjects (male and female) with one or more psychological symptoms. Intervention: Biofield energy treated proprietary capsule. Primary Outcome Measures: Psychological questionnaire scores and functional physiological biomarkers. Results: Psychological and mental health symptoms were statistically and significantly reduced in the subjects taking the Blessed TRI 360TM capsule compared to the placebo group on both days 90 and 180. Furthermore, levels of functional physiological biomarkers were higher in the intervention group compared to the placebo group: vitamins (C, B12, and D3 active metabolites), neurotransmitters (acetylcholine, noradrenaline, and dopamine), hormones (17-ß-estradiol, oxytocin, and insulin), antioxidant enzyme (catalase), and anti-aging protein (klotho). Additionally, proinflammatory cytokines (TNF-α, IL-1ß, IL-8, IFN-γ, and IL-2), progesterone, and oxidative stress markers (MDA and oxidized-LDL) were lower in the Blessed TRI 360TM group than the placebo group after the intervention. Conclusions: Altogether, the Blessed TRI 360TM dietary supplement capsule was found to be safe and tolerable. It significantly improved psychological symptoms and mental disorders and simultaneously improved different functional physiological biomarkers that led to an improvement in the overall health and quality of life of the adult subjects. Clinical Trial Registration Number: ECR/147/Inst/GJ/2013/RR-16.

Efficacy of a novel proprietary dietary supplement (TRI 360TM) on psychological symptoms and stress-related quality of life in adult subjects: A randomized controlled clinical trial.

Nowadays, diet plays an increasingly important role in normal physiology and mental health. Recently, many studies have shown that more use of dietary supplements in mental and psychological disorders. Study objective was to investigate safety and efficacy of proprietary nutraceutical combination (TRI 360TM) on psychological symptoms in adult human subjects with one or more psychological symptoms in open-label, single-center, parallel-group, randomized controlled trial. Eighty-four participants aged 20-45 years with psychological symptoms were completed this trial. Participants were randomly assigned to placebo and treatment groups. Treatment group received TRI 360TM capsules twice a day. TRI 360TM was well-tolerated and didn't show treatment-related adverse-events upto 180 days. All assessed perception scorings on psychological symptoms like fatigue, mental stress, sleep disturbance, anxiety, depression, emotional trauma, mood changes, self-confidence, willpower, and motivation were very significantly (p ≤ 0.0001) improved in TRI 360TM participants than placebo control group. Furthermore, significantly (p ≤ 0.001) increased levels of functional biomarkers: vitamin C and D3 metabolites, neurotransmitters, hormones, antiaging protein (klotho) level; and decreased proinflammatory cytokines and oxidative stress marker, malondialdehyde in TRI 360TM group than placebo. According to these findings, the use of TRI 360TM supplementation as a potentially safe therapeutic option for reducing psychological symptoms in healthy adults.

Cannabidiol improves thyroid function via modulating vitamin D3 receptor in vitamin D3 deficiency diet-induced rat model.

The study was evaluated the impact of cannabidiol (CBD) on thyroid hormones by modulation cannabinoid receptor-2 (CB2) and vitamin D receptor (VDR) in rats fed with vitamin D3 deficiency diet (VDD). CB2-receptors were analyzed by RT-PCR method and others biomarkers by ELISA. The relative expression of CB2 (thyroid ~ 4 folds), VDR protein (liver, 151.72%), and (kidney, 66%) was significantly increased in CBD-60 compared to VDD. Vitamin D3 metabolites were significantly increased serum (189.42%), kidney (73.84%), and liver (58.11%) in CBD-60 than VDD. Increased thyroxine (59.9%) and calcitonin (213.59%); while decreased thyroid-stimulating hormone (36.15%) and parathyroid hormone (38.64%) was observed CBD treatment in VDD rats. In conclusion, CBD treatment improves CB2 and VDR expression and the level of vitamin D3 metabolites, along with improved thyroid hormones, including calcitonin. This is the first report with an improved CB2 and VDR expression after CBD treatment in VDD induced animals. Thus, CBD can be considered to use in hypothyroidism conditions and to maintain bone health.

Anti-inflammatory potential of cannabidiol (CBD) on combination of caecal slurry, LPS, and E. coli-induced systemic inflammatory response syndrome (SIRS) in Sprague Dawley Rats.

OBJECTIVE: The study objective was to evaluate the therapeutic effect of cannabidiol (CBD) on a combination of caecal slurry, lipopolysaccharide (LPS), and Escherichia coli (E. coli)-induced systemic inflammatory response syndrome (SIRS) in male Sprague Dawley rats. METHODS: The therapeutic activity was monitored in behavioral tests and inflammatory biomarkers by the enzyme-linked immune sorbent assay (ELISA) method. RESULTS: Behavioral tasks were significantly increased like a tail flick response by 73.84% (p ≤ 0.001), grip strength by 33.56% (p ≤ 0.028), locomotor activity by 20.71% (p = 0.034) in the CBD (60 mg/kg) group compared to disease control (DC) group. Levels of inflammatory serum biomarkers like interleukin-1ß (IL-1ß), matrix metallopeptidase-9 (MMP-9), IL-6, and tumor necrosis factor-alpha (TNF-α) were significantly decreased by 29.56 (p = 0.041), 71.20 (p ≤ 0.001), 35.05 (p ≤ 0.001), and 75.56% (p = 0.002), respectively, in the CBD-60 compared with DC. Inflammatory cytokines levels, viz. IL-1ß, MMP-9, IL-6, and TNF-α, in the liver were significantly (p ≤ 0.001) decreased by 81.01, 40.41, 22.84, and 69.46%, respectively, in CBD-60 to DC. Similarly, levels of inflammatory cytokines such as IL-1ß and MMP-9 in the kidney were significantly (p ≤ 0.001) decreased by 80.90 and 43.93%, respectively, in CBD-60 compared to DC. CONCLUSION: Taken together, results suggest that CBD treatment significantly improved behavioral tasks and decreased the level of inflammatory cytokines under SIRS conditions that might provide an opportunity to manage acute and chronic inflammatory disorders.

CD155: A Key Receptor Playing Diversified Roles.

Cluster of differentiation (CD155), formerly identified as poliovirus receptor (PVR) and later as immunoglobulin molecule, is involved in cell adhesion, proliferation, invasion and migration. It is a surface protein expressed mostly on normal and transformed malignant cells. The expression of the receptor varies based on the origin of tissue. The expression of the protein is determined by factors involved in the sonic hedgehog pathway, Ras-MEK-ERK pathway and during stressful conditions like DNA damage response. The protein uses an alternate splicing mechanism, producing four isoforms, two being soluble (CD155ß and CD155γ) and two being transmembrane protein (CD155α and CD155δ). Apart from being a viral receptor, researchers have identified CD155 to play important roles in cancer research and the cell signaling field. The receptor is recognized as a biomarker for identifying cancerous tissue. The receptor interacts with molecules involved in the cells' defense mechanism. The immunesurveillance role of CD155 is being deciphered to understand the mechanistic approach it utilizes as an onco-immunologic molecule. CD155 is a non-MHC-I ligand which helps in identifying non-self to NK cells via an inhibitory TIGIT ligand. The TIGIT-CD155 pathway is a novel MHC-I-independent education mechanism for cell tolerance and activation of NK cells. The receptor also has a role in metastasis of cancer and transendothelial mechanism. In this review, the authors discuss the virus-host interaction that occurs via a single transmembrane receptor, the poliovirus infection pathway, which is being exploited as a therapeutic pathway. The oncolytic virotherapy is now a promising modality for curing cancer.

Solid and liquid state characterization of tetrahydrocurcumin using XRPD, FT-IR, DSC, TGA, LC-MS, GC-MS, and NMR and its biological activities.

Tetrahydrocurcumin (THC) is one of the major metabolites of curcumin (CUR), an ancient bioactive natural polyphenolic compound. This research article describes both the solid and liquid state characterization of THC using advanced spectroscopic and thermo-analytical techniques. Anti-inflammatory, anti-oxidant, and neuroprotective activities of THC were investigated using in vitro cell lines. Liquid chromatography-mass spectrometry analysis revealed that our sample comprised 95.15% THC, 0.51% tetrahydrodemethoxycurcumin (THDC), 3.40% hexahydrocurcumin, and 0.94% octahydrocurcumin. Gas chromatography-mass spectrometry analysis indicated the presence of 96.68% THC and 3.32% THDC. THC in solution existed as keto-enol tautomers in three different forms at different retention time, but the enol form was found to be dominant, which was also supported by nuclear magnetic resonance analysis. THC was thermally stable up to 335.55 °C. THC exhibited more suppression of cytokines (TNF-α, IL-1ß, and MIP-1α) than CUR in a concentration-dependent manner in mouse splenocytes, while NK-cell and phagocytosis activity was increased in macrophages. THC showed a significant reduction of free radicals (LPO) along with improved antioxidant enzymes (SOD and catalase) and increased free radical scavenging activity against ABTS+ radicals in HepG2 cells. THC displayed higher protection capability than CUR from oxidative stress and neuronal damage by improving cell viability against H2O2 induced HepG2 cells and MPP+ induced SH-SY5Y cells, respectively, in a concentration-dependent manner. Thus, a variation of the biological activities of THC might rely on its keto-enol form and the presence of other THC analogs as impurities. The present study could be advantageous for further research on THC for better understanding its physicochemical properties and biological variation.

A Validated LC-MS/MS Method for Simultaneous Determination of 3-O-Acetyl-11-Keto-ß-Boswellic Acid (AKBA) and its Active Metabolite Acetyl-11-Hydroxy-ß-Boswellic Acid (Ac-11-OH-BA) in Rat Plasma: Application to a Pharmacokinetic Study.

The aim of this study was to develop and validate a new, rapid, sensitive, selective and reliable liquid chromatography-tandem mass spectrometry method for simultaneous determination of 3-O-Acetyl-11-keto-ß-boswellic acid (AKBA) and its active metabolite 3-O-Acetyl-11-hydroxy-ß-boswellic acid (Ac-11-hydroxy-BA) in rat plasma. Both analytes (AKBA and Ac-11-hydroxy-BA) and the internal standard (IS, ursolic acid) were extracted from 100 µL of rat plasma by protein precipitation. Chromatographic separation was achieved on PRP-H1 RP-C18 column (75 mm × 2 mm, 1.6 µm) using acetonitrile-water (95.5 v/v) as the mobile phase. Mass detection was conducted by electrospray ionization in positive ion multiple reaction monitoring (MRM) mode. A linear dynamic range of 1-1,000 ng/mL for both AKBA and Ac-11-hydroxy-BA was established with mean correlation coefficient (r (1)) of 0.999. Intra- and inter-day precision (% CV) of analysis were found in the range of 1.9-7.4%. The accuracy determined for these analytes ranged from 92.4 to 107.2%. The extraction recoveries for both analytes ranged from 92.6 to 97.3% for spiked plasma samples and were consistent. The % change in stability samples compared to nominal concentration ranged from 0.4 to 4.2%. This method was successfully tested to a pharmacokinetic (PK) study for estimation of AKBA and acetyl-11-hydroxy-BA in rat plasma following oral administration of AKBA. This method has been validated with the advantage of shorter run time that can be used for high-throughput analysis and has been successfully applied to the pharmacokinetic study of AKBA in rats.

Investigation of Molecular Properties that Influence the Permeability and Oral Bioavailability of Major ß-Boswellic Acids.

BACKGROUND AND OBJECTIVES: Boswellic acids (BAs) include ß-boswellic acid (BA), 3-acetyl-ß-boswellic acid, 11-keto-ß-boswellic acid, 3-acetyl-11-keto-ß-boswellic acid, ß-boswellic alcohol, and 3-acetyl-11-hydroxy-ß-BA from Boswellia species, and are the main active ingredients of Boswellia serrata extracts (BSE). BSE have been used for the treatment of different inflammatory diseases; however, their pharmaceutical development has been severely limited by their poor oral bioavailability. The aims of this study were to investigate the molecular properties of six BAs, and to determine their experimental aqueous solubility, partition coefficient (Log P), gastrointestinal stability, adsorption-desorption kinetics, and permeability studies. METHODS: The physicochemical properties of six BAs were obtained from SMILES representations using ChemDraw, and MarvinSketch. The molecular properties were also determined experimentally. The permeability studies were performed using parallel artificial membrane permeability assay (PAMPA), and Caco-2 cells. RESULTS: The experimental Log P values of BAs correlated well (R2 = 0.94) with the calculated Log P values. Metabolic stability data confirmed that BAs were found to be unstable in simulated gastrointestinal fluids and intestinal S9 fractions. The apparent permeability (Papp) range of BAs in both PAMPA and Caco-2 for the apical (AP) to basolateral (BL) was in the range of 0.52 ± 0.05 × 10-6 to 2.84 ± 0.14 × 10-6cm/s. The efflux ratio of Papp (BL → AP) to Papp (AP → BL) for all BAs was < 2 in Caco-2 cells, suggesting greater permeability in the absorptive direction. Caco-2 cell adsorption studies confirmed the accumulation of BAs (35-55%) inside the enterocytes. These compounds exhibited a strong correlation between PAMPA and Caco-2 cell monolayer permeation data. CONCLUSIONS: The results of the present study have shown an empirical relationship between the molecular properties and intestinal absorption of BAs for the first time.

Betulinic Acid, the first lupane-type triterpenoid isolated via bioactivity-guided fractionation, and identified by spectroscopic analysis from leaves of Nyctanthes arbor-tristis: its potential biological activities in vitro assays.

Betulinic acid was first time isolated via bioactivity-guided fractionation from ethyl acetate extract of Nyctanthes arbor-tristis leaves. Its structure was established by FTIR, 1H and 13C- nuclear magnetic resonance and high-resolution mass spectrometry. It had shown excellent inhibition of anti-inflammatory properties with IC50 of 10.34 µg/mL (COX-1), 12.92 µg/mL (COX-2), 15.53 µg/mL (5-LOX), 15.21 µg/mL (Nitrite), 16.65 µg/mL (TNF-α), and also exhibited potent antioxidant activity with IC50 of 18.03 µg/mL. The anticancer activity of betulinic acid was evaluated against different human cancer cell lines. It showed significant cytotoxicity against various cancer cell lines with an IC50 of 6.53 (HepG2), 9.34 (A549), 14.92 (HL-60), 16.90 (MCF-7), 17.07 (HCT-116), 13.27 (PC-3), and 12.55 µM (HeLa). This is the first report on isolation and identification of the unreported lupane-type triterpenoid, betulinic acid from leaves of Nyctanthes arbor-tristis, which showed potent anti-inflammatory, antiproliferative, and antioxidant activity in vitro assays.

Role of Vital Trace Elements in Nanocurcumin-Centered Formulation: A Novel Approach to Resuscitate the Immune System.

The present paper described the immunomodulatory potential of novel nanocurcumin-based formulation enriched with trace elements and vitamins on cyclophosphamide-induced immunosuppression in rat model. Major immune-related assays were monitored such as hemagglutination assay, delayed-type hypersensitivity (DTH) reaction, cellular immune response, IgG, IgE, IgM, cerebrospinal fluid biomarkers, hematological study, antioxidant profile, and lipid biomarkers. Chemical characterization of novel formulation showed retention time (R t ) 18.98 of curcumin, while LC-MS data revealed the presence of the curcumin mass at m/z 369.01 [M + H]+ (calculated for C21H21O6+, 369.13). This novel formulation exhibited significantly (p ≤ 0.001) increased primary and secondary antibody titer by 72.41% and 33.25%, respectively, while DTH response being improved by 87.50% (p ≤ 0.01). However, CD4+, CD8+, and CD28+ counts were significantly (p ≤ 0.05) increased by 76.46%, 68.21%, and 19.29%, respectively, while the concentrations of IgE, IgM, and IgG were significantly (p ≤ 0.05) increased by 40%, 28.43%, and 38.75%, respectively. CSF biomarkers analysis showed a decreased level of corticosterone, dopamine, serotonin, and tau protein by 29.38%, 51.73%, 29.93%, and 4.87%, respectively. Antioxidant enzymes such as CAT, GPx, and SOD were increased by 43.74%, 49.00%, and 40.84%, respectively, and non-enzymatic component, GSH, was increased by 55.52%. Similarly, free radical LPO was significantly (p ≤ 0.05) decreased by 40.37%, and acute inflammatory marker, MPO concentration, was reduced by 31.14%, compared with the disease control group. In addition, supportive hematology and lipid profile analysis showed promising results with improved overall animal profile. Thus, trace elements in novel formulation can be used in the various pharmacological activities and as dietary supplement due to its wide properties.

Immunomodulatory potential of nanocurcumin-based formulation.

Vitamins, minerals, and nanocurcumin play a substantial role in various nutraceutical/pharmaceutical formulations that are widely used in therapeutics, cosmetics, and dietary supplements. The current study aimed to investigate the comparative in vitro immunomodulatory effect of a novel nanocurcumin-based formulation with curcumin in LPS-induced cytokine expression, NK cells' activity, and phagocytosis. The proinflammatory cytokines (TNF-α, IL-1ß, and MIP-1α) and NK cells' activity were measured in cell supernatants using ELISA assay; however, phagocytosis activity was performed using colorimetric analysis. The chemical characterization of novel nanocurcumin-based formulation using LC-MS (R t 19.02 min) and mass spectra analysis (m/z 369.04) confirmed the presence of the curcumin in highest peak concentration. MTT assay in three tested cell-lines showed that the formulation was found non-toxic at all the tested concentrations. The expression of TNF-α, IL-1ß, and MIP-1α in splenocytes was significantly (p ≤ 0.001) inhibited. Besides, the NK cells' activity and phagocytosis (macrophage) were increased significantly (p ≤ 0.001). Overall, the promising results of this study indicated the significant immunomodulatory effect of nanocurcumin-based formulation compared to the curcumin, which could be used against various inflammatory disorders such as allergy, asthma, autoimmune diseases, coeliac disease, inflammatory bowel disease, etc.

Effect of a Novel Ashwagandha-based Herbomineral Formulation on Pro-inflammatory Cytokines Expression in Mouse Splenocyte Cells: A Potential Immunomodulator.

BACKGROUND: Herbomineral formulations are momentous in an audience of worldwide by virtue of their holistic approach to life. These formulations are widely used as complementary therapies in immunocompromised patients including cancer. Still, there is the need of cost-effective and safe herbomineral-based formulation that can modulate immune response by the regulation of cytokines cascades. OBJECTIVE: Current study, we investigated immunomodulatory effect of TEBEH in LPS-induced cytokines expression levels in mouse splenocytes in vitro. MATERIALS AND METHODS: The most effective and safe concentrations of TEBEH were chosen by determining the cell viability of splenocytes using MTT assay. The pro-inflammatory cytokines such as TNF-α, IL-1ß, MIP-1α, and IFN-γ were measured in cell supernatants using ELISA. RESULTS: MTT data showed TEBEH formulation was found safe up to 10.53 µg/mL. At noncytotoxic concentrations (0.00001053-10.53 µg/mL), TEBEH significantly (P ≤ 0.001) inhibited the expressions of TNF-α, IL-1ß, and MIP-1α in mouse splenocytes as compared with vehicle control. CONCLUSION: In summary, TEBEH may indeed promote an anti-inflammatory environment by suppression of pro-inflammatory cytokines. These observations indicated that TEBEH has potential effects in downregulating the immune system and might be developed as a useful anti-inflammatory product for various inflammatory disorders. SUMMARY: The present study was undertaken to evaluate an immunomodulatory effect of the herbomineral formulation in LPS-induced mouse splenocytes with the measurement of cytokines expression such as TNF-α, IL-1ß, MIP-1α and IFN-γ. The results showed that the expression of TNF-α, IL-1ß, and MIP-1α was significantly down-regulated while, IFN-γ was significantly up-regulated in mouse splenocytes. It is hypothesized that modulation of the proinflammatory cytokines might occur via NF-κB pathway. Therefore, the herbomineral test formulation might act as an effective anti-inflammatory and immunomodulatory product, and this can be used as a complementary and alternative treatment for the prevention of various types of inflammatory and auto-immune disorders Abbreviations used: LPS: Lipopolysaccharide, IL: Interleukin; NF-κB: Nuclear factor kappa-B, TNF-α: Tumor necrosis factor alpha, MIP-1α: Macrophage inflammatory protein-1α, IFN-γ: Interferon, MTT: 3-(4,5-diamethyl-2-thiazolyl)-2, 5-diphenyl-2Htetrazolium), ELISA: Enzyme linked immune sorbent assay, ANOVA: Analysis of variance.