Efficacy of Bromelain along with Trypsin, Rutoside Trihydrate Enzymes and Diclofenac Sodium Combination Therapy for the treatment of TMJ Osteoarthritis - A Randomised Clinical Trial.

INTRODUCTION: Osteoarthritis (OA) is a degenerative disorder characterized by chronic inflammatory response of cartilage and articular surface involving Temporomandibular Joint (TMJ). Pain as one of the major symptom of osteoarthritis affects the quality of life and is usually managed by Non Steroidal Anti Inflammatory Drugs (NSAIDs) such as diclofenac sodium. Bromelain, trypsin and rutoside trihydrate formulation can be used to treat this disease because of anti-inflammatory and anti-oxidant effects. AIM: To assess the effectiveness of oral bromelain, trypsin, rutoside trihydrate enzymes and diclofenac sodium combination therapy over diclofenac sodium for the treatment of TMJ osteoarthritis. MATERIALS AND METHODS: Thirty Patients with symptomatic TMJ osteoarthritis were randomly divided into three groups. 10 patients were treated with diclofenac sodium (Group 1), 10 were given oral enzymes (bromelain, trypsin, rutoside trihydrate) and diclofenac sodium combination (Group 2), and 10 were treated with oral enzyme preparation (bromelain, trypsin, rutoside trihydrate) (Group 3). Patients were evaluated on day 1, day 4, day 7 and day 10. Comparison of pain rating within three groups was assessed using numeric rating scale. The efficacy criteria were analysed applying ANOVA followed by post-hoc test. RESULTS: Inter group comparison of the effectiveness of management of pain, resulted in a value p < 0.05 between Group 2 and other two groups, which indicated that Group 2 patients responded better than Group 1 and Group 3, while p > 0.05 between Group 1 and Group 3 showed both groups responded equally to the treatment. CONCLUSION: The trial showed significant improvement in reducing pain in patients treated with oral enzymes and diclofenac sodium combination therapy.

Anti inflammatory potential of Decalepis hamiltonii (Wight and Arn) as evidenced by down regulation of pro inflammatory cytokines-TNF-alpha and IL-2.

ETHNOPHARMACOLOGICAL RELEVANCE: Ancient tribes in the Western Ghats of India use the roots of Decalepis hamiltonii Wight and Arn (Asclepiadaceae) for several medicinal purposes particularly inflammation. AIM: To investigate whether the pure compounds obtained from the Decalepis hamiltonii have anti inflammatory activity. MATERIALS AND METHODS: The bioactive lead molecules from the roots of Decalepis hamiltonii were extracted into dichloromethane/methanol and purified by silica gel column chromatography. Structural elucidation of the purified compounds was performed with (1)H and (13)C NMR and mass spectrometry. The in vitro anti inflammatory activity of the pure compounds was studied in mitogen induced peripheral blood mononuclear cells (PBMCs) employing [(3)H] thymidine uptake assay and their effect on cytokine expression by reverse transcriptase polymerase chain reaction (RT-PCR). The inhibition of nuclear factor kappaB (NF-kappaB) activity in the presence of pure compounds was determined in J774 A.1 cells. The cytotoxicity of the compounds was tested using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay kit. RESULTS AND DISCUSSION: Lupeol acetate (Compound 1) and (2S)-5,7,4'-trihydroxy flavanone 4'-O-beta-d-glucoside (Compound 2) isolated from Decalepis hamiltonii roots inhibited the proliferation of mitogen induced PBMCs with an IC(50) value of 8 and 0.5mug/ml respectively. MTT assay revealed the compounds to be non-cytotoxic. Though, both the compounds down regulated the synthesis of mRNA of the pro inflammatory cytokines, interleukin-2 (IL-2) and tumour necrosis factor-alpha (TNF-alpha), the anti inflammatory cytokine interleukin-10 (IL-10), was found to be up regulated. NF-kappaB activation in J774 A.1 cells were also inhibited by both the compounds. CONCLUSION: Lupeol acetate and (2S)-5,7,4'-trihydroxy flavanone 4'-O-beta-d-glucoside isolated from Decalepis hamiltonii roots showed anti inflammatory activities by down regulating TNF-alpha and IL-2 specific mRNA, besides up regulating the synthesis of mRNA of IL-10.

Detection and quantification of phytotoxic metabolites of Sarocladium oryzae in sheath rot-infected grains of rice.

AIMS: The present study describes the detection and quantification of the Sarocladium oryzae metabolites, helvolic acid and cerulenin in extracts of rice grains collected from plants infected with sheath rot. It also describes the phytotoxicity of these metabolites on rice seedlings. METHODS AND RESULTS: Helvolic acid and cerulenin in sheath rot-infected rice grains were detected using thin layer chromatography (TLC) and nuclear magnetic resonance (NMR) analyses. On the TLC plates helvolic acid and cerulenin moved as brownish yellow spots and showed R(F) values of 0.61 and 0.49, respectively. A standard assay curve was developed on the basis of selective toxicity of helvolic acid towards Calvibacter michiganensis ATCC 2140 and cerulenin towards Candida albicans 1150. The amounts of helvolic acid and cerulenin on the basis of standard assay curve were 2.2 and 1.75 microg g(-1) of infected seeds. Treatment of IR 36 rice seedlings with metabolites induced chlorosis and reduced shoot length by 20%, root length by 30% and root number by 7% relative to control. CONCLUSIONS: Helvolic acid and cerulenin were detected in infected rice grains and these metabolites induced chlorosis and reduced the seed viability and seedling health of rice. SIGNIFICANCE AND IMPACT OF THE STUDY: Antimicrobial and phytotoxic metabolites, helvolic acid and cerulenin are present in infected grains and reduce the seed viability and seedling health. These metabolites may increase the pathogenic potential and survival of S. oryzae in rice seed by competing with other seed-borne fungi.