Anusha Bhaskar

GC-MS analysis of phytocomponents in the methanolic extract of Eupatorium triplinerve

Objective: To characterize the phytochemical constituents of Eupatorium triplinerve using GC – MS. Methods: Ten grams of the powdered sample was subjected to column chromatography over silica gel (100- 200 mesh) and eluted with n-hexane, chloroform, ethanol and methanol respectively. n-hexane and chloroform did not elute much of the compounds. The methanol fraction of the Eupharbatum triplinerve was taken for GC-MS analysis. The analysis was carried out on a GC Clarus 500 GC system with a column packed with Elite – 1 (10% dimethyl poly siloxane, 30 x 0.25 mm ID x 1 EM df), the compounds are separated using with Helium as carrier gas at a constant flow 1ml/min. sample extract (2 μL) injected into the instrument was detected by Turbo gold mass detector (Perkin Elmer) with the aid of the Turbo mass 5.1 software. Results: The GC MS analysis provided peaks of eleven different phytochemical compounds namely hexadecanoic acid (14.65%), 2,6,10-trimethyl,14-ethylene-14-pentadecne (9.84%), Bicyclo[4.1.0]heptane, 7-butyl- (2.38%), Decanoic acid, 8-methyl-, methyl ester (3.86%), 1-undecanol (7.82%), 1-hexyl-1-nitrocyclohexane (2.09%), 1,14-tetradecanediol (6.78%), Octadecanoic acid, 2-hydroxy-1,3-propanediyl ester (19.18%) and 2-hydroxy-3-[(9E) -9-octadecenoyloxy] propyl(9E)-9-octadecenoate (8.79%). Conclusion: The bioactive compounds in the methanolic extract of Eupatorium triplinerve have been screened using this analysis. Isolation of individual components would however, help to find new drugs.

Evaluation of the wound-healing activity of Hibiscus rosa sinensis L (Malvaceae) in Wistar albino rats

Objective: To investigate the wound-healing potency of the ethanolic extract of the flowers of Hibiscus rosa sinensis. Materials and Methods: The wound-healing activity of H. rosa sinensis (5 and 10% w/w) on Wistar albino rats was studied using three different models viz., excision, incision and dead space wound. The parameters studied were breaking strength in incision model, granulation tissue dry weight, breaking strength and collagen content in dead space wound model, percentage of wound contraction and period of epithelization in excision wound model. The granulation tissue formed on days 4, 8, 12, and 16 (post-wound) was used to estimate total collagen, hexosamine, protein, DNA and uronic acid. Data were analyzed by Analysis of Variance (ANOVA) test. P<0.05 was considered statistically significant. Results: The extract increased cellular proliferation and collagen synthesis at the wound site, as evidenced by increase in DNA, total protein and total collagen content of granulation tissues. The extract-treated wounds were found to heal much faster as indicated by improved rates of epithelialization and wound contraction. The extract of H. rosa sinensis significantly (P<0.001) increased the wound-breaking strength in the incision wound model compared to controls. The extract-treated wounds were found to epithelialize faster, and the rate of wound contraction was significantly (P<0.001) increased as compared to control wounds. Wet and dry granulation tissue weights in a dead space wound model increased significantly (P<0.001). There was a significant increase in wound closure rate, tensile strength, dry granuloma weight, wet granuloma weight and decrease in epithelization period in H. rosa sinensis-treated group as compared to control and standard drug-treated groups. Conclusion: The ethanolic extract of H. rosa sinensis had greater wound-healing activity than the nitrofurazone ointment.

Antihyperglycemic, antioxidant and hypolipidemic effect of Punica granatum L flower extract in streptozotocin induced diabetic rats

Abstract Objective To investigate the antihyperglycemic, hypolipidemic and antioxidant effects of aqueous and ethanolic flower extracts of Punica granatum (P. granatum) in streptozotocin (STZ) induced diabetes mellitus in wistar rats. Methods Antihyperglycemic, hypolipidemic and antioxidant effect of P. granatum aqueous (PGFAet) and ethanolic extracts (PGFEet) at doses of 200 and 400 mg/kg bw was evaluated in streptozotocin induced diabetic rats. Glycosylated hemoglobin, plasma insulin, cholesterol and lipoprotein levels, lipid peroxide levels and the antioxidant enzyme levels were determined. Results The administration of the extracts markedly reduced blood glucose and glycosylated hemoglobin increased the levels of plasma insulin and liver glycogen. The extract also had a hypolipidemic activity decreasing the levels of total cholesterol, LDL-cholesterol, VLDL-cholesterol and triglycerides. The levels of lipid peroxides in terms of thiobarbituric acid reactive substances (TBARS) were remarkably reduced and the activities of the enzymes SOD, CAT and GPx and GSH were increased. We also saw an increase in the activity of the enzyme glucokinase and decrease in glucose-6-phosphatase. It can be concluded that the flower extracts of P. granatum (PGFAet and PGFEet) has significant restorative effect on the blood glucose, hyperlipidemia and oxidative stress. Conclusions It can be concluded from the studies that the ethanolic extracts of the flowers of P. granatum at dosage 400 mg/kg/day exhibit significant effect in lowering blood sugar and lipid levels and increasing the antioxidant enzymes when compared to the aqueous extract. Glycosylated hemoglobin levels were reduced and the extract exhibited a stimulatory effect on insulin.

A database for medicinal plants used in treatment of asthma

The knowledge of most plants used in the treatment of asthma, the plant part which is effective in treatment is confined to very few persons who are engaged in folklore medicine. However, this form of medicine is not very popular. Therefore, it is of considerable interest to ethno-botanical community to understand the plants and the parts used for treatment. Here, we describe AsthmaPlantBase, a database containing information of medicinal plants for treatment of asthma. Availability http://www.asthmaplants.com.

Proteomic analysis of plasma proteins in diabetic retinopathy patients by two dimensional electrophoresis and MALDI-Tof-MS

OBJECTIVE Diabetic retinopathy is a highly specific vascular complication of diabetes mellitus and progresses from mild non-proliferative abnormalities characterized by increased vascular permeability to moderate and severe proliferative diabetic retinopathy characterized by the growth of blood vessels on the retina. The aim of the study was to identify the differentially expressed proteins in diabetic retinopathy using two-dimensional electrophoresis. METHODS Blood sample was drawn from subjects with diabetes mellitus (without retinopathy) who served as controls and patients with diabetic retinopathy in tubes containing EDTA as anticoagulant. Albumin and immunoglobulin IgG collectively removed to enrich proteins of lower abundance. 2de was carried out to see if there are any differentially expressed proteins. RESULTS Approximately 48 and 61 spots were identified in control and diabetic retinopathy respectively, of which three protein spots RBP1 (retinol-binding protein 1), NUD10 (Diphosphoinositol polyphosphohydrolase 3 alpha), NGB (neuroglobin) were down regulated and HBG2 (hemoglobin) and BY55 (CD 160 antigen) were upregulated in diabetic retinopathy. These five protein spots were excised and were subjected to in-gel tryptic digestion, and their identities were determined by ultraflex MALDI-TOF-MS. CONCLUSION We report a comprehensive patient-based plasma proteomic approach to the identification of potential biomarkers for diabetic retinopathy screening and detection. SIGNIFICANCE OF THE STUDY We identified 5 different proteins that were differentially expressed in the plasma of control diabetic patients (without retinopathy). Among these five proteins the expression of neuroglobin (NGB) protein varied significantly and may be a potential biomarker in diabetic retinopathy.

Prophylactic efficacy of S-2(2-aminoethylamino)ethyl phenyl sulfide (DRDE-07) against sulfur mustard induced lung toxicity in mice

Abstract Objective: The present study was planned to investigate the prophylactic efficacy of S-2(2-aminoethylamino)ethyl phenyl sulfide (DRDE-07), against topically applied SM induced pulmonary toxicity in mouse. Materials and methods: Animals were pretreated with S-2(2-aminoethylamino)ethyl phenyl sulfide (DRDE-07) (249.4 mg/kg by oral gavage) 30 minutes before SM exposure. The SM (6.48 mg/kg) was applied on hair clipped dorsocaudal region (percutaneous) of the animal. The animals were sacrificed on day 1, 3, 5 and 7. The biochemical changes those were observed in the bronchoalveolar lavage (BAL) fluid and lung tissue included protein, LDH, MPO, β-glucuronidase, MMP-2, MMP-9, activated macrophages, reduced glutathione and lipid peroxidation level. Results and discussion: Pretreatment with DRDE-07 (0.2 LD50) attenuated SM-induced changes at all time point tested. BAL fluid biochemical endpoints indicated epithelial and endothelial cell damages as evidenced by increase in BAL protein, LDH level and increased number of activated macrophages. The increased myeloperoxidase activity and β-glucuronidase level exhibited the degranulation of neutrophils due to SM toxicity in lung. The zymogrphy analysis of BAL fluid showed a significant increase in matrix metalloproteases (MMP) activity due to inflammatory cells accumulation. Conclusion: Thirty minutes pretreatment with DRDE-07 decreased vascular permeability reduced the inflammation and oxidative stress, hence may be recommended as a potential prophylactic agent for SM intoxication.