Evaluation of immunomodulatory and antioxidant properties of traditional Kwath, conventional extracts of plants Cocculus hirsutus and Cuscuta reflexa - in vitro &ex vivo studies.

INTRODUCTION: The consumption of 'Patalagarudi' (Cocculus hirsutus 'CHP') and 'Amarbel' (Cuscuta reflexa 'CRA') as ethnic plants for health promotions rarely validated. The limited literature reported these plants as antioxidant and immunomodulators. OBJECTIVE: To evaluate the biodynamic properties of CHP and CRA extracts. METHODOLOGY: The traditional formulation, 'Kwath' (K) and conventional extracts were prepared with CRA and CHP. The total phenolic content (TPC) was estimated. Various polyphenol compounds in the extracts were eluted on UHPLC. The biodynamic activities; i. Free radical scavenging (FRS-DPPH and ABTS), ii. Intracellular ROS scavenging activity in RAW 264.7 cell line iii. Spleenocytes proliferation assay for Th1/Th2 Immunomodulatory potential by flow-cytometer were assessed. RESULTS: The TPC in CRA (105-159 µg GAE/mg) and CHP (35-48 µg GAE/mg) recorded. The chromatographic peaks confirmed the presence of polyphenols in CRA and CHP extracts. UV spectra of the extracts to the extent possible have been correlated with certain polyphenols. The FRS (IC50) was significantly low in CRA-K (DPPH = 22.7; ABTS = 12.0 µg/ml) than CHP-K (DPPH = 70.4; ABTS = 50.2 µg/ml). Similarly, intracellular ROS scavenging activity with CRA-K (84%) showed the highest inhibitory potential compared to CHP-K (50%) and LPS control. The immunomodulatory activity of CRA-K significantly upregulated TH1 cytokines (TNFα and IFN-γ). The downregulation of Th2 cytokines (IL-4 and IL-10) was in all CRA and CHP extracts as compared to Con A. CONCLUSION: The current study confirms the immunomodulatory and antioxidant properties of CRA and CHP along with the presence of polyphenols.

Comparative evaluation of nephroprotective potential of resveratrol and piperine on nephrotic BALB/c mice.

OBJECTIVE: The objective of this study was to evaluate the nephroprotective potential of resveratrol and piperine at same dose on cationic bovine serum albumin (cBSA) induced immune complex glomerulonephritis (ICGN) in BALB/c mice. MATERIALS AND METHODS: Female BALB/c mice were divided into five groups. Group I served as normal control (complete Freund's adjuvant + Saline). Two weeks later, Groups II, III, IV, and V were administered cBSA (13 mg/kg) via the caudal vein 3 times/week every alternative day for 6 weeks to induce ICGN. Simultaneously, from the 3rd week, Groups III, IV were treated with resveratrol and piperine up to 6 weeks. Group V was treated with methylprednisolone considered as a reference standard. RESULTS: There was a significant decrease in albuminuria, serum creatinine, and blood urea nitrogen in Group IV animals when compared with Group III. In addition, Group III and IV have comparable results with cBSA treated animals. Concurrently, same groups showed significantly comparable variance in antioxidant enzymes, phagocytic index, and neutrophil adhesion assay. Group IV found to be more significant in IgG1 reduction than Group III. CONCLUSION: The findings of this study well-demonstrated that piperine has potential immunomodulatory and anti-inflammatory activity than resveratrol; therefore, piperine needs special attention in autoimmunity and inflammation research.

Nitrogen doped nanocrystalline semiconductor metal oxide: An efficient UV active photocatalyst for the oxidation of an organic dye using slurry Photoreactor.

Water pollution is a cause for serious concern in today's world. A major contributor to water pollution is industrial effluents containing dyes and other organic molecules. Waste water treatment has become a priority area in today's applied scientific research as it seeks to minimize the toxicity of the effluents being discharged and increase the possibility of water recycling. An efficient and eco-friendly way of degrading toxic molecules is to use nano metal-oxide photocatalysts. The present study aims at enhancing the photocatalytic activity of a semiconductor metal oxide by doping it with nitrogen. A sol-gel cum combustion method was employed to synthesize the catalyst. The prepared catalyst was characterized by FT-IR, XRD, UV-DRS, FESEM and AFM techniques. UV-DRS result showed the catalyst to possess band gap energy of 2.97eV, thus making it active in the UV region of the spectrum. Its photocatalytic activity was evaluated by the degradation of a model pollutant-Orange G dye, under UV light irradiation. Preliminary experiments were carried out to study the effects of pH, catalyst dosage and initial dye concentration on the extent of dye degradation. Kinetic studies revealed that the reaction followed pseudo first order kinetics. The effect of electrolytes on catalyst efficiency was also studied. The progress of the reaction was monitored by absorption studies and measuring the reduction in COD. The catalyst thus prepared was seen to have a high photocatalytic efficiency. The use of this catalyst is a promising means of waste water treatment.

Lead and trace element levels in placenta, maternal and cord blood: a cross-sectional pilot study.

AIM: To determine lead (Pb) and trace element (Fe, Zn, Cu, Mg) levels in placenta, maternal and cord blood; further, to assess the interactions between Pb and trace elements, if any. METHODS: A cluster of 'at term pregnant women' from rural and urban areas (n = 30 each, total n = 60) as well as their neonates after delivery were recruited. Maternal blood, heel prick neonatal blood, placenta and cord blood were collected at the time of parturition. Pb and trace element levels in blood/serum, placenta were determined on graphite furnace atomic absorption (AAS) and flame AAS respectively. Due to sample constraints, only Pb levels were determined in heel prick blood samples of neonates. RESULTS: There were no major abnormal signs and symptoms, however, 38% had pale conjunctiva and 13% had pigmented tongue. The blood lead levels (BLL) were significantly (P < 0.05) higher in urban post-partum women compared to their rural counterparts. Higher BLL did not correlate with either pregnancy outcome or neonatal anthropometry. Maternal serum trace element levels were deficient in both rural and urban women. Significant (P < 0.01) positive correlation between maternal and neonatal BLL as well as significant (P < 0.05) inverse correlation between cord BLL and placental Pb levels were observed. Maternal and cord BLL correlated inversely (P < 0.05) with maternal and cord serum Fe levels, respectively. CONCLUSION: Urban post-partum women and their neonates were at higher risk with elevated BLL. Estimate of cord blood Pb may not be the true index of neonatal BLL. These observations must be confirmed in a larger cohort because prenatal/neonatal screening avoids the risks associated with rest of life.

Lead and trace element levels in milk and blood of buffaloes (Bubalus bubalis) from Hyderabad, India.

The presence of lead (Pb) in milk and its interaction with trace elements is a serious health concern. Present study is aimed at determining Pb and trace element (Fe, Zn and Mg) levels in milk and blood/serum samples of lactating buffaloes (Bubalus bubalis) living in a market-area (Group-A) and a dairy-experimental station (Group-B), Hyderabad, India. In addition, kidney and liver function tests were assessed. Fodder, milk and blood Pb levels were significantly (p < 0.01) higher in Group-B. Elevated Pb levels correlated positively with reduced Fe and Zn levels in both serum and milk. A significant (p < 0.01) positive correlation between blood Pb and milk Pb levels was observed. Kidney and liver function markers were significantly higher in Group-B buffaloes. The results suggest that contaminated fodder might be one of the responsible factors for elevated Pb levels. In addition, lower levels of Fe and Zn might have led to bioaccumulation of Pb in blood and milk.

Pre-clinical toxicity & immunobiological evaluation of DNA rabies vaccine & combination rabies vaccine in rhesus monkeys (Macaca mulatta).

BACKGROUND & OBJECTIVES: Pre-clinical toxicology evaluation of biotechnology products is a challenge to the toxicologist. The present investigation is an attempt to evaluate the safety profile of the first indigenously developed recombinant DNA anti-rabies vaccine [DRV (100 µg)] and combination rabies vaccine [CRV (100 µg DRV and 1.25 IU of cell culture-derived inactivated rabies virus vaccine)], which are intended for clinical use by intramuscular route in Rhesus monkeys. METHODS: As per the regulatory requirements, the study was designed for acute (single dose - 14 days), sub-chronic (repeat dose - 28 days) and chronic (intended clinical dose - 120 days) toxicity tests using three dose levels, viz. therapeutic, average (2x therapeutic dose) and highest dose (10 x therapeutic dose) exposure in monkeys. The selection of the model i.e. monkey was based on affinity and rapid higher antibody response during the efficacy studies. An attempt was made to evaluate all parameters which included physical, physiological, clinical, haematological and histopathological profiles of all target organs, as well as Tiers I, II, III immunotoxicity parameters. RESULTS: In acute toxicity there was no mortality in spite of exposing the monkeys to 10XDRV. In sub chronic and chronic toxicity studies there were no abnormalities in physical, physiological, neurological, clinical parameters, after administration of test compound in intended and 10 times of clinical dosage schedule of DRV and CRV under the experimental conditions. Clinical chemistry, haematology, organ weights and histopathology studies were essentially unremarkable except the presence of residual DNA in femtogram level at site of injection in animal which received 10X DRV in chronic toxicity study. No Observational Adverse Effects Level (NOAEL) of DRV is 1000 ug/dose (10 times of therapeutic dose) if administered on 0, 4, 7, 14, 28 th day. INTERPRETATION & CONCLUSIONS: The information generated by this study not only draws attention to the need for national and international regulatory agencies in formulating guidelines for pre-clinical safety evaluation of biotech products but also facilitates the development of biopharmaceuticals as safe potential therapeutic agents.

Effect of Mangiferin and Mahanimbine on Glucose Utilization in 3T3-L1 cells.

BACKGROUND: Stem barks of Mangifera indica contain a rich content of mangiferin (xanthone glucoside), whereas Murraya koenigii leaves contain rich sources of mahanimbine (carbazole alkaloid) and used traditionally for the treatment of diabetes. OBJECTIVE: To investigate the effects of mangiferin (xanthone glucoside) and mahanimbine (carbazole alkaloid) on glucose utilization in 3T3-L1 cells. MATERIALS AND METHODS: Mangiferin was isolated from stem barks of Mangifera indica and mahanimbine was isolated from Murraya koenigii leaves. These isolated compounds were subjected to MTT assay and glucose utilization test with 3T3-L1 cells. RESULTS: Treatment of the 3T3-L1 cells with mangiferin and mahanimbine increased the glucose utilization in a dose-dependent manner. At a concentration of 1 mM, mangniferin showed 2-fold increase in glucose utilization compared with untreated control. In case of mahanimbine, the observed effect at 1 mM was almost equivalent to positive control (insulin at 1 µM). Moreover, MTT assay showed that both of these compounds were less toxic at a concentration of 1 mM (nearly 75% cells are viable). CONCLUSION: The present results indicated that these natural products (mangiferin and mahanimbine) exhibited potential ethnomedical uses in management of diabetes.

Nonclinical toxicology study of recombinant-plasmid DNA anti-rabies vaccines.

The absence of standard guidelines from National and International regulatory agencies for the safety evaluation of biotechnology products challenges the ingenuity of toxicologists. At present, the development of standard pre-clinical toxicology protocols for such products is on an individual case basis. The present investigation is an attempt to evaluate the safety profile of the first indigenously developed DNA based anti-rabies vaccine in India. The test compounds were DNA rabies vaccine [DRV (100 microg)] and combination rabies vaccine (CRV (100 microg DRV and 1/50 dose of cell culture vaccine)), intended for clinical use by intramuscular route on 1, 7, 14 and 28 day. As per the regular mandatory requirements, the study has been designed to undertake acute (single dose--10 days), sub-chronic (repeat dose--28 days) and chronic (intended clinical dose--120 days) toxicity tests using three dose levels viz. therapeutic, average (2 x therapeutic dose) and highest dose (10 x therapeutic dose) exposure in Swiss Albino mice. The selection of the rodent model viz. Swiss Albino mice is based on affinity and rapid higher antibody response during the efficacy studies. Apart from physical, physiological, clinical, hematological and histopathology profiles of all target organs, the tier-I immunotoxicity parameters have also been monitored. There were no observational adverse effects even at levels of 10x therapeutic dose administration of DRV and CRV. The procedure also emphasizes on the designing of protocols for the products developed by recombinant technique.

Studies on the development of colon-targeted delivery systems for celecoxib in the prevention of colorectal cancer.

The nonsteroidal anti-inflammatory drugs (NSAIDs) are found to be potential chemopreventive agents of colorectal cancer. Celecoxib, an NSAID with selective cyclooxygenase-2 inhibition, was proved to be effective for the prevention of colon cancer in patients with familial adenomatous polyposis (FAP) and sporadic polyps. In the light of this information, the present study was carried out to develop oral colon-targeting drug delivery systems for celecoxib using guar gum as a carrier. Matrix tablets containing various proportions of guar gum were prepared by wet granulation technique using starch paste as a binder. The tablets were evaluated for hardness, drug content and were subjected to in vitro drug release studies. The amount of celecoxib released from the matrix tablets at different time intervals was estimated by a HPLC method. Guar gum matrix tablets released only 2-4% of celecoxib in the physiological environment of stomach and small intestine depending on the proportion of guar gum used in the formulation. When the dissolution study was continued in simulated colonic fluids (rat caecal content medium), the matrix tablets containing 20% of guar gum released another 37% of celecoxib after degradation by the colonic bacterial action. The matrix tablets containing 30% of guar gum released about 24% of celecoxib in simulated colonic fluids indicating the susceptibility of the guar gum formulations to the rat caecal contents. The results of the study show that the matrix tablets containing either 20 or 30% of guar gum are most likely to target celecoxib for local action in the colon. The guar gum matrix tablets of celecoxib showed no change either in physical appearance, drug content or in dissolution pattern after storage at 40 degrees C/RH 75% for 6 months. Differential scanning calorimetry (DSC) studies indicated no possibility of interaction between celecoxib and guar gum/other formulation excipients.