EVALUATION OF IN VITRO- ANTI-OXIDANT POTENTIAL OF AQUEOUS ROOT EXTRACT OF CLERODENDRUM SERRATUM L.

Objective: The intent  of this report  was to investigate the effect of aqueous root extract of Clerodendrum serratum L. for antioxidant activity using divergent models viz. DPPH scavenging assay, Superoxide scavenging assay and Ferric Reducing Antioxidant Power (FRAP) assay.Materials and Methods: The root of C. serratum was extracted using water. The yield of aqueous extract was 10%w/w. The outcome was examined statistically by the regression method.Results and discussions: The IC50 values are 85.43 µg/ml and 107.59 µg/ml for DPPH radical scavenging and Superoxide scavenging assay respectively whereas  FRAP showed significant reducing power activity with increased concentration of sample. The pilot study showed, a significant correlation existed between concentrations of the extract and percentage engrossment of free radicals.Conclusion: The antioxidant property may be corresponding to the polyphenols and flavonoids adjacent in the extract. These results clearly revealed that C. serratum might be effective against diseases analogous with free radical mediated. Keywords Clerodendrum serratum, DPPH, Superoxide, FRAP, Rutin, Antioxidan

ANTIOXIDANT, ANTI-INFLAMMATORY, AND ANTIDIABETIC ACTIVITY OF HYDROALCOHOLIC EXTRACT OF OCIMUM SANCTUM: AN IN-VITRO AND IN-SILICO STUDY

Objective: Ocimum sanctum is known to be beneficial in the management of diabetes, however, its mechanism remains unexplored. This study wasaimed to gain insite into the mechanisms through which it may counter diabetes and its complications.Methods: Hydroalcoholic whole plant extract of O. sanctum was screened for its antidiabetic potential and ability to counter oxidative andinflammatory stress through various in-vitro assays. Further, bioactive compounds that may be responsible for its antidiabetic activity were predictedthrough molecular-docking studies.Results: Crude extractive yield of 35.43% was obtained from Soxhlet extraction which mainly showed the presence of flavonoids, alkaloids, glycosides, and saponins. Plant extract showed good potential to scavenge 2,2-diphenyl-1-picrylhydrazyl free radical (40.95-68.71%) which may be attributed\ to its high phenolic (0.366 mg gallic acid equivalent/g) and flavonoid (0.113 mg quercetin equivalent/g) contents. Plant showed exceptional antiinflammatory activity which was evaluated through inhibition of protein denaturation (47.61-82.37%) and red blood cell membrane stabilization assay (43.66-78.28%). Further, extract treatment greatly inhibited α-glucosidase enzyme (34.17-71.45%) but failed to produce noticeable inhibition of α-amylase activity (1.94-14.88%). Docking studies predicted that rosmarinic acid, stigmasterol, linalool, bieugenol, and aesculin may be responsible for antidiabetic activity possessed by the plant through their interaction with the insulin receptor.Conclusion: These findings conclude that O. sanctum may be beneficial in managing diabetes and its associated complications through inhibitingα-glucosidase activity, reducing oxidative and inflammatory stress.Keywords: Diabetes mellitus, Ocimum sanctum, Antioxidant, Anti-inflammatory, Docking, Diabetic complications

FORMULATION AND PHYSICOCHEMICAL EVALUATION OF NANOSTRUCTURED LIPID CARRIER FOR CODELIVERY OF CLOTRIMAZOLE AND CIPROFLOXACIN

Objective: The purpose of this research was to formulate nanostructured lipid carrier system (NLCs) in such a way that they can be applied forbacterial as well as fungal infectious diseases.Methods: To achieve the prime objective, varying concentrations of clotrimazole (CLZ) and ciprofloxacin (CIPRO) were selected for formulations.Stearic acid (solid lipid polymer), oleic acid (liquid lipid polymer), and polyvinyl alcohol (surfactant) were utilized for formulating NLCs throughsolvent diffusion technique. NLCs were characterized for their surface morphology, Fourier transform infrared spectroscopy (FTIR) drug-polymerinteraction, particle size distribution, zeta potential, loading capacity, drug entrapment efficacy (EE), and in vitro drug release profile.Results: NLCs were fabricated with size range varying from 276 nm to 564 nm, possessing smooth spherical morphology. No drug-polymer interactionwas observed through FTIR analysis. The highest drug EE for CLZ and CIPRO was found to be 78.6% and 65.8%, respectively. Formulated NLCs depictthe biphasic release profile with initial burst release of 40% within 2 hrs followed by controlled release.Conclusion: Better homing of drug molecules and controlled drug release through formulated NLCs makes them suitable carrier system for variousanti-microbial and anti-fungal applications.Keywords: Nanostructured lipid carries, Clotrimazole, Ciprofloxacin, Solvent diffusion method, In vitro release

HYDROALCOHOLIC EXTRACTION OF SHOOT CULTURES FROM NOTHAPODYTE NIMMONIANA AND ITS ANTI-PROLIFERATIVE ANALYSIS

Objective: The important restraints in plant-derived medicine are the convenience of active composites which depends on diverse topographical situations, active compound build-up pattern, environmental circumstances and genetic makeup of the plant. The presented work was aimed to describe an efficient technique for fast propagation of Nothapodytes nimmoniana which is an imperative source of Camptothecin.Methods: Multiple shoots were successfully regenerated from the explants N. nimmoniana using Murashige and Skoog (MS) medium accompanied with IBA+KN+GA3 (1:3:2) mg/l. the hydro-alcoholic extract was obtained by soxhlet extraction for field grown roots (fgRE), shoots (fgSE) and in vitro plantlets (ivPL) and further subjected to phytochemicals analysis. HPLC analysis was performed to determine the percentage of camptothecin (CAM). Anti-proliferative studies were carried out followed by Acridine orange/Ethidium bromide staining. Extracts were also estimated for reactive nitrogen species (RNS) and reactive oxygen species (ROS).Results: Preliminary qualitative chemical analysis of the extracts displayed the presence of alkaloids, carbohydrates, steroids, saponins, terpenoids and phenolics compounds. HPLC analysis revealed the presence of CAM in all the extracts. ivPL exhibited lowest anti-proliferative activity in contrast to other extract and percentage of ROS and RNS was also less. Although ivPL did not exhibited prominent anti-tumour effect, but in vitro accumulation of CAM in 2-months old shoots provides an alternative resource to 5 y-old field grown plants.Conclusion: This work delivers a potential to lead to further increase the potency of CAM in in vitro shoots of two months old as an alternative to the field grown plants for treating cancers

IN VITRO AND IN SILICO EVALUATION OF THE ANTIDIABETIC EFFECT OF HYDROALCOHOLIC LEAF EXTRACT OF CENTELLA ASIATICA

Objective: Aim of the present study was to explore the possible mechanisms through which Centella asiatica may be beneficial in managing diabetes and its associated complications by in vitro methods and to predict the potential bioactive constituent/s responsible for its anti-diabetic activity through in silico docking study.Methods: Hydro-alcoholic leaf extract of C. asiatica was prepared using Soxhlet extraction. Plant extract was evaluated for its in vitro antioxidant, anti-inflammatory and anti-diabetic activity. Further, docking screening was performed using Molegro Virtual Docker software to predict potential moiety which may be responsible for its anti-diabetic activity.Results: Soxhlet extraction resulted in extractive yield of 35.43% and showed high antioxidant potential as demonstrated by its ability to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (98.72%). This antioxidant activity may be attributed to high phenolic and flavonoid components present in extract (1.004 mg gallic acid equivalent (GAE)/gram and 0.113 mg Rutin equivalent (RE)/gram dried extract respectively). Plant extract inhibited albumin denaturation (81.77%) and stabilized RBCs membrane (66.7%) indicating its high anti-inflammatory potential. In vitro anti-diabetic assays revealed that anti-hyperglycaemic activity of this plant can be attributed to its high efficiency to inhibit α-amylase (62.13%) and glucosidase (59.9%) enzymatic activity, which are well established targets for the management of diabetes. Further, through docking studies we predicted that centellasaponin-C, asiaticoside, asiaticoside-E, castilliferol and brahminoside present in this plant might be responsible for the anti-diabetic properties exhibited by this plant.Conclusion: These results provide a scientific justification for the traditional anti-diabetic use of this plant. It may control diabetes through lowering dietary glucose uptake and may benefit in progression of diabetic complications through reducing oxidative and inflammatory stress. Predicted anti-diabetic molecules need to be screened further for the management of hyperglycemia

QUALITY AND AFFORDABILITY OF AMOXICILLIN GENERIC PRODUCTS: A PATIENT CONCERN

Objective: Antibiotics save millions of lives from infectious diseases worldwide. Failing treatment, serious adverse effects, and antimicrobial resistance are constantly reported mainly from developing nations due to lack of quality of antibiotics medicines. In India quality of medicines remains a major regulatory challenge and patients concern. Thus, a pilot study to explore the quality of generic amoxicillin products and associated price along with their burden on patients was evaluated.Methods: 46 amoxicillin trihydrate generic products with the label claim of 250 mg amoxicillin were procured from open market of Northern India without prescription. Identification and quantitative evaluations of these generic products were estimated using Indian Pharmacopoeia (IP) 2010 recommended High-Performance Liquid Chromatography (HPLC) method. And assay value was compared with the maximum retail price per unit dosage. Affordability was estimated in general for the population who daily live on less than Indian rupees Rs. 144 and `88.6.Results: Out of 46 products, 28.26% were found to be out of IP specification, including 13.04% products which were of substandard quality. Fishers' exact test with p-value 0.87 showed products quality gaps were irrespective of their price. Ceiling price of Rs. 3.04 per unit dosage pose a high burden on the patients who are on amoxicillin treatment.Conclusion: Situation demands the evidence of safety before approval and thereafter too. However, the situation may become worst if the price and quality could not be controlled. Thus Indian drug regulatory bodies need to be entreated to counter these critical issues.Â

SOLID LIPID NANOPARTICLE: FABRICATED THROUGH NANOPRECIPITATION AND THEIR PHYSICOCHEMICAL CHARACTERIZATION

Objective: Aim of the present study was to formulate solid lipid nanoparticles (SLNs) and to determine their physicochemical parameters when stored at cold temperature in aqueous solution (D-SLNs) prior to biological application.Methods: SLNs were formulated though nanoprecipitation technique which comprised of stearic acid (lipid), poloxamer 188 and lecithin (surfactant). Physicochemical parameters were estimated though particle size analysis, polydispersity index, surface morphology analysis (Scanning electron microscopy and Transmission electron microscopy) and cytotoxicity studies followed by live-dead staining through acridine orange and ethidium bromide.Results: SLNs with spherical morphology were successfully fabricated as revealed though SEM and TEM investigations. Fabricated SLNs had the mean particle size ranging from 188 nm (SLNs) to 327 nm (D-SLNs). Zeta potential was found to be±14mV to±6mV and polydispersity index was 0.297±0.18 for SLNs without incubation and 0.538±0.07 for SLNs after incubation. No cytotoxicity was observed for SLNs.Conclusion: SEM and TEM investigations showed morphological variation in SLNs and D-SLNs. Dissimilarity in mean particle size, zeta potential and polydispersity index indicates the increase in size and aggregation of nanoparticles. No cytotoxic effects of SLNs were observed in normal cells, suggesting storage of nanoformulation in the aqueous state has no effect in context to cytotoxicity. Hence we conclude that prolonged storage of formulation at cold temperature causes the deterioration of polymeric formulation

ANTIFERTILITY ACTIVITY AND CONTRACEPTIVE POTENTIAL OF THE HYDROALCOHOLIC RHIZOME EXTRACT OF TRILLIUM GOVANIANUM IN FEMALE WISTAR RATS

Objective: Trillium govanianum is used in several traditional containing steroids and sex hormones for the management of inflammation, menstrual disorders, sex-related disorders, and antiseptic. The present study was aimed to investigate the antifertility potential of hydroalcoholic rhizome extract of T. govanianum and to explore the possible mechanism of action. Methods: Anti-implantation activity of T. govanianum rhizome extract (125 and 250 mg/kg; p.o.) was performed in female Wistar rats with proven fertility, and its estrogenic/antiestrogenic effect was evaluated in ovariectomized females. 17-α-ethinylestradiol (1 μg/rat/day; s.c.) or plant extract was administered for 11 days after which animals were sacrificed. Percentage inhibition of implantation sites, serum estrogen levels, changes in body and uterus weight, and morphological alterations in the uterus and ovaries were evaluated. Results: T. govanianum treatment resulted in increased uterus weight and induced dose-dependent anti-implantation effect, with 100% implantation inhibition at 250 mg/kg dose. Anti-implantation effects of T. govanianum were associated with endometrial thickening and significantly elevated serum estrogen levels. Moreover, estrogenic/antiestrogenic studies revealed that T. govanianum possessed strong estrogenic effect; however, the effect was saturable. Conclusion: T. govanianum possesses antifertility activity which can be attributed to its strong estrogenic potential and uterine thickening. Moreover, it could find a clinical application as a safer and efficacious birth control herbal remedy

EFFICIENT HYDROALCOHOLIC EXTRACTION FOR HIGHEST DIOSGENIN CONTENT FROM TRILLIUM GOVANIANUM (NAG CHHATRI) AND IT'S IN VITRO ANTICANCEROUS ACTIVITY

ABSTRACTObjective: The present study involves hydroalcoholic extraction of Trillium govanianum (Hindi name: Nag chhatri), which is a high-value medicinalplant found at the altitude of 2500-4000 m. Aiming in this direction, we performed hydrolysis of extract using response surface methodology (RSM)for optimizing diosgenin content. The extracts were evaluated for cytotoxicity.Methods: In RSM, the cumulative effect of independent variables including time (minutes), temperature (°C), and solid-liquid ratio (g/ml)were investigated through central composite design. Cytotoxicity studies of crude rhizome and hydroalcoholic extract were carried out by3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on three cell lines, viz., MDCK, MCF-7, and MDA-MB-23.Results: The diosgenin content obtained was 5.9%, which is reported for the first time in T. govanianum. Hydrolyzed extract showed less toxicity inMDCK (normal cell line) cells but significantly reduced the proliferation of MCF-7 and MDA-MB-231 cancer cells.Conclusion: Hydrolysis method was optimized by RSM, which proved to be an efficient method for extraction of diosgenin from T. govanianum.Hydrolyzed extract showed antiproliferative activity on cancer cell lines with minimal effect on normal cells.Keywords: Trillium govanianum, Diosgenin, Nag chhatri, Hydrolysis, MDCK, MCF-7, MDA-MB-231

Effect of Ca2EDTA on zinc mediated inflammation and neuronal apoptosis in hippocampus of an in vivo mouse model of hypobaric hypoxia.

Calcium overload has been implicated as a critical event in glutamate excitotoxicity associated neurodegeneration. Recently, zinc accumulation and its neurotoxic role similar to calcium has been proposed. Earlier, we reported that free chelatable zinc released during hypobaric hypoxia mediates neuronal damage and memory impairment. The molecular mechanism behind hypobaric hypoxia mediated neuronal damage is obscure. The role of free zinc in such neuropathological condition has not been elucidated. In the present study, we investigated the underlying role of free chelatable zinc in hypobaric hypoxia-induced neuronal inflammation and apoptosis resulting in hippocampal damage.Adult male Balb/c mice were exposed to hypobaric hypoxia and treated with saline or Ca2EDTA (1.25 mM/kg i.p) daily for four days. The effects of Ca2EDTA on apoptosis (caspases activity and DNA fragmentation), pro-inflammatory markers (iNOS, TNF-α and COX-2), NADPH oxidase activity, poly(ADP ribose) polymerase (PARP) activity and expressions of Bax, Bcl-2, HIF-1α, metallothionein-3, ZnT-1 and ZIP-6 were examined in the hippocampal region of brain.Hypobaric hypoxia resulted in increased expression of metallothionein-3 and zinc transporters (ZnT-1 and ZIP-6). Hypobaric hypoxia elicited an oxidative stress and inflammatory response characterized by elevated NADPH oxidase activity and up-regulation of iNOS, COX-2 and TNF-α. Furthermore, hypobaric hypoxia induced HIF-1α protein expression, PARP activation and apoptosis in the hippocampus. Administration of Ca2EDTA significantly attenuated the hypobaric hypoxia induced oxidative stress, inflammation and apoptosis in the hippocampus.We propose that hypobaric hypoxia/reperfusion instigates free chelatable zinc imbalance in brain associated with neuroinflammation and neuronal apoptosis. Therefore, zinc chelating strategies which block zinc mediated neuronal damage linked with cerebral hypoxia and other neurodegenerative conditions can be designed in future