Acute and sub chronic toxicity studies with herbal pain relieving formula (Rhuleave-K™) in rats.

Rhuleave-K™ is a proprietary combination of Curcuma longa extract, Boswellia serrata extract and black sesame seed oil. Acute toxicity was evaluated as per OECD guidelines 423. Rhuleave-K™ was fed at 2000 mg/kg to overnight fasted female rats. Clinical signs of abnormality and mortality was observed daily for 14 days. Sub-chronic toxicity was studied by feeding Rhuleave-K™ at 100, 500 and 1000 mg/kg/day to rats as per OECD guidelines 408. After 90 days feeding, hematological and biochemical parameters were analyzed. Histopathology of all the major organs was also studied. In the acute toxicity study, there was no clinical sign of toxicity in any of the rats at maximum dose of 2000 mg/kg. The LD50 was computed as >2000 mg/kg in rats. The repeated dosing of Rhuleave-K™ at the maximum dose level of 1000 mg/kg for 90 days did not induce any observable toxic effects in rats, when compared to its corresponding control. The hematology and biochemistry profiles of treated rats were similar to control animals and difference was non-significant (p > 0.05). The histopathology of major organs of all the control and treated animals was normal. In this study the NOAEL for Rhuleave-K™ was calculated as 1000 mg/kg daily in rats.

Dendrobium chrysanthum ethanolic extract induces apoptosis via p53 up-regulation in HeLa cells and inhibits tumor progression in mice.

Background Dendrobium is one of the diverse genus of orchid plants. It possesses a number of pharmacological activities and has long been used in traditional system of medicine. The goal of this study was to investigate the apoptosis inducing property of the ethanolic extract from the leaves of Dendrobium chrysanthum, a species of Dendrobium whose anticancer role has not been ascertained yet. Methods To evaluate the anticancer activity of the ethanolic extract of D. chrysanthum in vitro in HeLa (human cervical cancer) cells, cytotoxic activity, generation of reactive oxygen species (ROS), induction of apoptosis and effect on cell cycle were determined. The in vivo study was carried out in Dalton's lymphoma (DL) bearing mice to assess the tumor growth delay. Results Our study demonstrated that the ethanolic extract showed dose-dependent cytotoxicity against HeLa cells. The extract exhibited dose-dependent increase in ROS production as well as apoptotic cell death which was further confirmed through presence of DNA fragmentation. Cell cycle analysis by flow cytometry suggests that the ethanolic extract perturbed cell cycle progression and leads to the delay of the cells in S phase. Further, the real-time PCR studies also showed up-regulation of apoptotic genes p53 and Bax. The in vivo antitumor activity exhibited significant increase in the life span of DL bearing mice as compared to control with significant decrease in abdominal size along with reduced tumor ascites. Conclusions These observations demonstrate the anticancer potential of the D. chrysanthum ethanolic extract mediated through p53-dependent apoptosis.

Clinical presentations, risk factors and outcomes of ceftazidime-resistant Gram-negative endophthalmitis.

BACKGROUND: To describe the clinical features and outcomes of patients diagnosed with ceftazidime-resistant Gram-negative endophthalmitis and the role of intravitreal imipenem in these cases. DESIGN: Retrospective consecutive interventional case series at a tertiary eye care centre in South India. PARTICIPANTS: Consecutive cases of ceftazidime-resistant Gram-negative endophthalmitis from April 2010 to December 2014. Fifty-six cases diagnosed during this time period were included. METHODS: All cases were managed with vitreous biopsy/vitrectomy, microscopy and undiluted vitreous culture, antimicrobial susceptibility of bacterial isolates and received intravitreal antibiotics. MAIN OUTCOME MEASURES: Anatomic and visual outcome of these cases, antimicrobial susceptibility pattern of intravitreal imipenem and outcome of cases injected with it. RESULTS: Commonest presentation was acute endophthalmitis following cataract surgery (27 eyes, 48.21%). Pseudomonas aeruginosa was isolated in 33 eyes (58.93%; 95% CI 46.05-71.81%). Nineteen eyes (34%; 95% CI 21.59-46.41%) developed phthisis; 14 eyes (25%; 95% CI 13.66-36.34%) had vision <20/200; 17 eyes (30.35%; 95% CI 18.31-42.39%) eyes had an ambulatory vision >20/200 (logMAR 1); 6 eyes (10.71%; 95% CI 2.61-18.81%) had a reading vision >20/40 (logMAR 0.3). Trend was towards better anatomic (72.73% vs. 40%) (P = 0.05) and visual improvement in the imipenem group (logMAR 3.94 + 0.21 to 2.43 + 1.4; P = 0.002), as compared with non-imipenem group (logMAR 2.99 + 1.3 to 2.55 + 1.4; P = 0.13). CONCLUSIONS: Outcome of ceftazidime-resistant Gram-negative endophthalmitis is poor. P. aeruginosa is the commonest isolated organism. All cases were sensitive to imipenem. There was a trend towards better anatomic outcome in imipenem-treated eyes.

Development and characterization of phyto-vesicles of wedelolactone for hepatoprotective activity.

Wedelolactone is isolated from the dried leaves of Eclipta alba (L.) and reported to be effective as a potential hepatoprotective, antibacterial and anti hemorrhagic. Pharmacokinetic studies of wedelolactone reveal its poor absorption through the intestine. The objective of the present study is to enhance bioavailability of wedelolactone by its complexation with phosphatidyl choline and then formulating it as phyto-vesicles for hepatoprotective activity. The complex of wedelolactone rich fraction was prepared with phosphatidyl choline and characterized on the basis of solubility, melting point, thin layer chromatography (TLC), UV, IR and NMR spectroscopy. The complex was further converted into phyto-vesicles and characterized. The hepatoprotective potential of phyto-vesicles was compared with complex, wedelolactone rich fraction and physical mixture of wedelolactone rich fraction and phosphatidyl choline by in vitro method. The results revealed that hepatoprotective activity is better in case of phyto-vesicles as compared to the complex, physical mixture and the wedelolactone itself. Enhanced bioavailability of the wedelolactone complex may be due to the amphiphillic nature of the complex, which greatly enhance the water and lipid solubility of the compound. The present study clearly indicates the superiority of phyto-vesicles over the complex and wedelolactone, in terms of better absorption and improved hepatoprotective activity.

Absorption enhancement of grape seed polyphenols by complexation with phosphatidyl choline.

Grape seed polyphenols (GPP) are reported to have various biological effects along with strong antioxidant potential. Pharmacokinetic studies of GPP reveal its poor absorption through the intestine. The objective of the present study was to enhance bioavailability of GPP by its complexation with phosphatidyl choline. A complex of GPP was prepared with phosphatidyl choline and characterized on the basis of solubility, melting point, DSC, and IR. Everted intestine sac technique was used to study ex vivo drug absorption of GPP-PC complex and plain GPP. Pharmacokinetic studies were performed in rats and the hepatoprotective activity of GPP-PC complex was also compared with GPP and GPP-PC physical mixture in isolated rat hepatocytes. Analytical reports along with spectroscopic data revealed the formation of the complex. The results of ex vivo study show that the GPP-PC complex has significantly increased absorption compared with GPP, when given in equimolar doses. The complex showed enhanced bioavailability, improved pharmacokinetics, and increased hepatoprotective activity as compared to GPP or GPP-PC physical mixtures. Enhanced bioavailability of GPP-PC complex may be due to the amphiphilic nature of the complex, which greatly enhance the lipid miscibility of GPP. The present study clearly indicates the superiority of complex over GPP, in terms of better absorption, enhanced bioavailability, and improved pharmacokinetics.

Development and evaluation of vesicular system for curcumin delivery.

Creams and gels containing curcumin are popularized worldwide and marketed all over the world, but even after incorporation of high amount of curcumin in topical formulations, significant antioxidant and anti-aging effect could not be achieved. Objective of the present study was to develop vesicular system for delivery of curcumin to achieve enhanced topical bioavailability. Complex of curcumin with phosphatidyl choline (PC) was prepared and characterized on the basis of TLC, DSC, melting point and IR spectroscopic analysis. The complex was further converted into vesicles (phyto-vesicles). Liposomes and niosomes of curcumin were also prepared and all these vesicular formulations were incorporated into carbopol gel to make feasible for topical application on skin. Anti-aging effects of these formulations were compared with plain curcumin and physical mixture of curcumin with phosphatidyl choline in UV-induced oxidative stress in mice. Analytical reports along with spectroscopic data revealed the formation of the complex. In the present study, the phyto-vesicles were found to be most effective than all other formulations and plain curcumin in providing enhanced antioxidant and antiaging effect. This increase may be due to the amphiphilic nature of the complex, which greatly enhances the water and lipid miscibility of the curcumin. This study clearly indicates the superiority of CU-PC complex and the phyto-vesicles prepared from CU-PC complex over others in providing enhanced anti-aging, antioxidant and anti-wrinkle effect.

Complexation with phosphatidyl choline as a strategy for absorption enhancement of boswellic acid.

Boswellic acids (BAs) are isolated from oleo gum resin of Boswellia serrata and are reported to be effective as anti-inflammatory, hypolipidemic, immunomodulatory, and anti-tumor. Pharmacokinetic studies of boswellic acid reveal its poor absorption through the intestine. The objective of the present study is to enhance bioavailability of boswellic acid by its complexation with phosphatidylcholine. A complex of boswellic acid was prepared with phosphatidylcholine and characterized on the basis of solubility, melting point, TLC, and IR. An everted intestine sac technique was used to study ex-vivo drug absorption of boswellic acid-phosphatidylcholine (BA-PC) complex and plain boswellic acid. Anti-inflammatory activity of the complex was compared with boswellic acid in carrageenan-induced paw edema in rats. Hypolipidemic activity was also evaluated in Triton-induced hyperlipidemia. The complex was also converted into vesicles (phytosomes) and compared with other vesicular systems (liposomes and niosomes) by evaluating its anti-inflammatory effect. Analytical reports along with spectroscopic data revealed the formation of a complex. The results of ex-vivo study show that BA-PC complex has significantly increased absorption compared with boswellic acid, when given in equimolar doses. The complex showed better anti-inflammatory and hypolipidemic activity as compared to BA. Among all vesicular systems phytosomes showed maximum anti-inflammatory activity. Enhanced bioavailability of the BA-PC complex may be due to the amphiphilic nature of the complex, which greatly enhance the water and lipid solubility of the boswellic acid. The present study clearly indicates the superiority of complex over boswellic acid, in terms of better absorption, enhanced bioavailability and improved pharmacokinetics.

Hepatoprotective activity of Cleome viscosa Linn. extract against thioacetamide-induced hepatotoxicity in rats.

Cleome viscosa Linn. (Family Capparidaceae) is naturalised throughout the hot and moist parts of India. Fresh leaves of this plant are used very effectively for the treatment of jaundice in the folk medicines of the Bundelkhand region of India. The hepatoprotective activity of the ethanolic extract of leaves was investigated against thioacetamide-induced hepatotoxicity in rats. The test material was found to be effectively hepatoprotective, as evidenced by biochemical parameters and histopathological studies. The hepatoprotective effect of ethanolic extract was comparable to that of silymarin, a standard hepatoprotective agent. The results of the present study support the traditional beliefs of the hepatoprotective effects of C. viscosa Linn.