A standardized black pepper seed extract containing ß-caryophyllene improves cognitive function in scopolamine-induced amnesia model mice via regulation of brain-derived neurotrophic factor and MAPK proteins.

ß-caryophyllene (BCP), a natural sesquiterpene present in plants, is a selective agonist of cannabinoid receptor type-2 (CB2) of the endocannabinoid system. In this study, we have prepared an extract from Piper nigrum (black pepper) seeds using supercritical fluid extraction, standardized to contain 30% BCP (ViphyllinTM ). The beneficial effects of prophylactic treatment with Viphyllin on cognitive functions were demonstrated in Scopolamine-induced dementia model mice. Male Swiss albino mice (25-30 g) were administered with Viphyllin (50 mg and 100 mg/kg body weight p.o.) or donepezil (1.60 mg/kg) for 14 days. Subsequently, cognitive deficits were induced by treating the animals intraperitoneally with Scopolamine (0.75 mg/kg). The cognitive behavior of mice was evaluated using a novel object recognition test (NORT) and Morris water maze (MWM) test. The brain homogenates were studied for biochemical parameters including cholinesterase activities and antioxidant status. Western blot analysis was performed to investigate the mechanism of action. Viphyllin dose dependently improved the recognition and spatial memory and cholinergic functions in Scop-treated mice. The extract was found protective against Scop-induced oxidative damage and histopathologic changes in the brain. At 100 mg/kg Viphyllin markedly reduced the proBDNF/mBDNF ratio (p < .05) and augmented the TrkB expression (p < .01). Viphyllin (100 mg/kg) was found to be neuroprotective by reducing the Scop-induced upregulation of p-JNK and p-p38 MAPK proteins, Bax/Bcl-2 ratio, and caspase activation in the brain. Viphyllin also exerted anti-inflammatory effects by downregulating Cox-2, TNF-α, and NOS-2 in Scop-induced mice (p < .05). To summarize, our data encourage Viphyllin as a functional ingredient/dietary supplement for brain health and cognition. PRACTICAL APPLICATIONS: Black pepper is a culinary spice having several medicinal attributes. Essential oils in the seeds of the plant give aroma and flavor to it. Here we have prepared an extract from the seeds of black pepper using supercritical fluid extraction, characterized for the presence of ß-caryophyllene (not <30%). This research work further validates the neuroprotective mechanism of the extract in Scopolamine-induced cognitive impairment model mice. The findings from this study strongly suggest the beneficial neuroactive properties of black pepper seed extract having the presence of BCP, a CB2 receptor agonist. It can thus be used potentially as a functional food ingredient for cognition and brain function.

Supplementation of green coffee bean extract in healthy overweight subjects increases lean mass/fat mass ratio: A randomized, double-blind clinical study.

OBJECTIVE: In this study, we have demonstrated that supplementation of a complex of chlorogenic acid isomers (CGA-7TM) could significantly mitigate the risk of obesity in healthy overweight subjects. METHOD: In a double-blind, placebo-controlled clinical study, healthy overweight (body mass index ⩾ 25 to <30 kg/m2) male and female subjects (N = 71) were randomly allocated to receive 500 mg CGA-7 or placebo daily for 12 weeks. Changes in body weight and body mass index were recorded alongside vital signs and anthropometric measurements at week 4, 8 and 12. Body composition was assessed at baseline and the end of treatment using dual-energy X-ray absorptiometry. Safety analysis included serum biochemical and haematological assessments and measurement of vital signs. In addition, any adverse or serious adverse events were recorded during the study. RESULTS: Sixty subjects completed the study. Mean body weight and body mass index were significantly reduced in CGA-7 group as compared to placebo (p < 0.001). CGA-7 group showed significant changes in body fat (%), fat mass and lean mass in comparison with placebo group (1.38% ± 1.4% vs -0.22% ± 0.86%, 1.97 ± 1.44 kg vs -0.39 ± 1.31 kg; 0.81 ± 1.20 kg vs -0.13 ± 0.97 kg, p < 0.001). Consumption of CGA-7 significantly improved the serum lipid profile. Importantly, CGA-7 consumption in humans had no adverse effects and was well tolerated during the study. The blood biochemical and haematological parameters marginally varied in the treatment groups throughout the study. CONCLUSION: To conclude, this study provides scientific validation of the functionality of green coffee bean extract and recommends the safety of the supplementation in healthy individuals.

Molecular docking analysis of Withaferin A from Withania somnifera with the Glucose regulated protein 78 (GRP78) receptor and the SARS-CoV-2 main protease.

Design and development of an effective compound to combat COVID-19 is clearly critical in the current circumstances. Therefore, it is of interest to document the molecular docking analysis data of the cellular receptor Glucose regulated protein 78 (GRP78) with Withaferin A from Withania somnifera in the context of COVID-19 pandemic for further consideration. Here, we report the optimal interaction features of withaferin A, artemisinin, curcumin and andrographolide with the GRP78 receptor having low binding energies (-8.7, -7.89, -6.21 and -6.17 kcal/mol respectively) in this report. In order to gain additional insights, the interaction pattern of compounds with SARS-CoV-2 main protease (Mpro) was studied.

A double blind, placebo-controlled randomized comparative study on the efficacy of phytosterol-enriched and conventional saw palmetto oil in mitigating benign prostate hyperplasia and androgen deficiency.

BACKGROUND: The present clinical trial was conducted to evaluate the efficacy and tolerability of a standardized saw palmetto oil containing 3% ß-sitosterol in the treatment of benign prostate hyperplasia (BPH) and androgen deficiency. METHODS: Subjects aged 40-65 years with symptomatic BPH were randomized to 12-week double-blind treatment with 500 mg doses of ß-sitosterol enriched saw palmetto oil, conventional saw palmetto oil and placebo orally in the form of capsules (n = 33 in each group). BPH severity was determined using the International Prostate Symptom Score (IPSS), uroflowmetry, serum measurement of prostate specific antigen (PSA), testosterone and 5α-reductase. During the trial, the androgen deficiency was evaluated using Aging Male Symptoms (AMS) scale, the Androgen Deficiency in the Aging Male (ADAM) questionnaire, serum levels of free testosterone. RESULTS: Subjects treated with ß-sitosterol enriched saw palmetto oil showed significant decrease in IPSS, AMS and ADAM scores along with reduced postvoiding residual volume (p < 0.001), PSA (p < 0.01) and 5α-reductase from baseline to end of 12-week treatment as compared to placebo. There was also a significant increment in the maximum and average urine flow rate (p < 0.001), and serum free testosterone level of subjects treated with enriched saw palmetto oil as compared to placebo. CONCLUSION: This study demonstrates the efficacy of ß-sitosterol enriched saw palmetto oil superior to conventional oil thus extending the scope of effective BPH and androgen deficiency treatment with improved quality of life through the intake of functional ingredients. TRIAL REGISTRATION: CTRI/2018/12/016724 dated 19/12/2018 prospectively registered. URL: http://ctri.nic.in/Clinicaltrials/advsearch.php.

Viwithan, a Standardized Withania somnifera Root Extract Induces Apoptosis in Murine Melanoma Cells.

BACKGROUND: Withania somnifera is an Indian medicinal herb known for the multipotential ability to cure various therapeutic ailments as described in the ayurvedic system of medicine. OBJECTIVE: In the present study, we have evaluated the antiproliferative activity of a standardized W. somnifera root extract (Viwithan) against different human and murine cancer cell lines. MATERIALS AND METHODS: The cytotoxicity of Viwithan was determined using thiazolyl blue tetrazolium blue assay and crystal violet staining. The apoptotic changes in B16F1 cells following treatment with Viwithan were observed by acridine orange/ethidium bromide (AO/EB) staining and DNA fragmentation assay. The binding affinity of withanolides in Viwithan with antiapoptotic proteins B-cell lymphoma 2, B-cell lymphoma-extra large, and myeloid cell leukemia 1 (MCL-1) were studied using in silico approach. RESULTS: The half maximal inhibitory concentration (IC50) values of Viwithan against liver hepatocellular carcinoma, Henrietta Lacks cervical carcinoma cells, human colorectal carcinoma cell line, and Ehrlich ascites carcinoma cells were 1830, 968, 2715, and 633 µg/ml, respectively. Interestingly, Viwithan was highly effective against B16F1 cells with an IC50 value of 220 µg/ml after 24 h treatment. The morphological alterations of apoptotic cell death were clearly observed in the AO/EB-stained cells after treatment with Viwithan. Viwithan induced late apoptotic changes in treated B16F1 cells as evident by the ladder formation of fragmented DNA in a time-dependent manner. The findings of molecular docking showed that withanolides present in Viwithan have a more binding affinity with the antiapoptotic proteins, particularly MCL-1. CONCLUSION: We have reported for the first time that Viwithan with 5% withanolides has a potent cytotoxic effect, particularly against B16F1 murine melanoma cells among the different cancer cell lines tested. SUMMARY: The present study reports for the first time that Viwithan, a standardized 5% Withania somnifera root extract, has potent cytotoxicity against B16F1 murine melanoma cellsWe have investigated the in vitro cytotoxicity of Viwithan in different human and murine cancer cells. Interestingly, we found that Viwithan was particularly very effective against B16F1 melanoma cells with a half maximal inhibitory concentration value of 220 µg/mlThe microscopic observations following acridine orange/ethidium bromide staining and DNA fragmentation assays clearly indicated that Viwithan might initiate late apoptosis in B16F1 cellsThe binding affinity of withanolides in Viwithan with antiapoptotic proteins of B-cell lymphoma 2 family was predicted using AutoDock tool. The results from in silico studies indicated a plausible synergistic effect of withanolides attributing to the Viwithan-induced apoptosis through suppression of intrinsic pathway for carcinogenesis. Abbreviations used: MTT: Thiazolyl blue tetrazolium blue; DMSO: Dimethyl sulfoxide; BSA: Bovine serum albumin; DMEM: Dulbecco's minimum essential medium; NCCS: National Centre for Cell Science; PBS: Phosphate-Buffered Saline; HepG2: Liver hepatocellular carcinoma; HeLa: Henrietta Lacks cervical carcinoma cells; HCT-116: Human colorectal carcinoma cell line; EAC: Ehrlich ascites carcinoma cells; IC50: Half maximal inhibitory concentration; AO/EB: Acridine orange/Ethidium bromide; BCL-2: B-cell lymphoma 2; BCL-XL: B-cell lymphoma-extra large; MCL-1: Myeloid cell leukemia 1; PDB: Protein Data Bank; ANOVA: Analysis of variance.

Bioavailability-enhanced Resveramax™ modulates quorum sensing and inhibits biofilm formation in Pseudomonas aeruginosa PAO1.

Quorum sensing (QS), a cell-to-cell communication mechanism present in many bacterial species plays a key role in regulating the virulence factor and biofilm formation in many pathogens, which causes severe public health risk. Therefore, interfering with QS mechanism offers an attractive strategy to combat bacterial infections. In the present study, anti-QS activity of a novel resveratrol formulation, Resveramax™, was detected using Chromobacterium violaceum biosensor bioassay and the effect of Resveramax on QS-regulated phenotypes in Pseudomonas aeruginosa PAO1 was assessed by standard protocols. Molecular docking analysis of resveratrol, the major active constituent of Resveramax, with LasR receptor protein was performed to evidence the QS-inhibitory potential of Resveramax. Results showed that Resveramax specifically inhibited the QS-mediated violacein pigment production in C. violaceum; pyocyanin production, proteolytic activity, swarming motility and biofilm formation in P. aeruginosa PAO1 in a concentration-dependent manner. Biofilms treated with Resveramax showed increased susceptibility to antibiotics when compared with the antibiotic treatment alone. Molecular docking analysis proved that resveratrol binds more rigidly with LasR receptor protein with docking score of -8.55 kJ/mol. These findings suggest that Resveramax could be used as novel QS-based antibacterial/anti-biofilm agent for the management of bacterial infections.

Synthesis, antimicrobial, DNA-binding and photonuclease studies of cobalt(III) and nickel(II) Schiff base complexes.

New metal complexes of the type [M(nih)(L)](PF(6))(n)·xH(2)O and [M(nih)(2)](PF(6))·xH(2)O (where M=Co(III) or Ni(II), L=1,10-phenanthroline (phen)/or 2,2' bipyridine (bpy), nih=2-hydroxy-1-naphthaldehyde isonicotinoyl hydrazone, n=2 or 1 and x=3 or 2) have been synthesized and characterized by elemental analysis, magnetic, IR and (1)H NMR spectral data. The electronic and magnetic moment 2.97-3.07 B.M. data infers octahedral geometry for all the complexes. The IR data reveals that Schiff base (nih) form coordination bond with the metal ion through azomethine-nitrogen, phenolic-oxygen and carbonyl-oxygen in a tridentate fashion. In addition, DNA-binding properties of these six metal complexes were investigated using absorption spectroscopy, viscosity measurements and thermal denaturation methods. The results indicated that the nickel(II) complex strongly bind with calf-thymus DNA with intrinsic DNA binding constant K(b) value of 4.9×10(4) M(-1) for (3), 4.2×10(4) M(-1) for (4), presumably via an intercalation mechanism compared to cobalt(III) complex with K(b) value of 4.6×10(4) M(-1) (1) and 4.1×10(4) M(-1) (2). The DNA Photoclevage experiment shows that, the complexes act as effective DNA cleavage agent.

Synthesis and biological evaluation of new Tetra-aza macrocyclic scaffold constrained oxadiazole, thiadiazole and triazole rings.

A new series of N,N'-(benzene-1,3-diyldi-1,3,4-oxadiazole-5,2-diyl)bis{2-[(5-benzene-1,3-diyl-1,3,4-oxadiazol-2-yl)amino]acetamide}(macrocycle 1), N,N'-(benzene-1,3-diyldi-1,3,4-thiadiazole-5,2-diyl)bis{2-[(5-benzene-1,3-diyl-1,3,4-thiadiazol-2-yl)amino]acetamide} (macrocycle 2) and S,S'-[benzene-1,3-diylbis(4H-1,2,4-triazole-5,3-diyl)]bis{[(5-benzene-1,3-diyl-4H-1,2,4-triazol-3-yl)sulfanyl]ethanethioate}(macrocycle 3) was synthesized from isophthalic dihydrazide (4) through a multistep reaction sequence. All the synthesized compounds were screened for their inhibitory effect against four different bacterial strains: P. aeruginosa ATCC-20852, K. pneumoniae MTCC-618, S. aureus ATCC- 29737, S. typhi MTCC- 3214. The synthesized compounds showed a significant zone of inhibition and the results were comparable with that of the standard drug ciprofloxacin. The synthesized compounds were further studied for their possible in vitro antioxidant effects by DPPH scavenging, total antioxidant capacity, total reductive capacity and H(2) O(2) scavenging activity. The results indicated that the in vitro antioxidant activity for all the three molecules was efficient when compared to the standards. The DNA interaction behavior of macrocycles 1-3 with CT-DNA was investigated by the absorption spectra obtained (K(b) constant for 1 is 4.53 × 10(4) M(-1) , for 2 is 5.75 × 10(4) M(-1) and for 3 is 5.86 × 10(4) M(-1) ). Based on the results it can be interpreted that the reducing power effect of the newly synthesized compounds demonstrates a direct effect on DNA binding and hence inhibiting the bacterial growth through their action on DNA by inhibiting DNA replication or DNA transcription.