DOXORUBICIN INDUCED COGNITION IMPAIRMENT IN RAT MODEL

 Objective: Doxorubicin (DOX) is a chemotherapeutic agent commonly used to treat a variety of cancers despite having well documented adverse sideeffects in organs like heart, liver, and kidney. The damage caused to brain, if any, and to what extent is, however, still not clear and the present studywas undertaken to estimate the possible role of DOX inducing cognitive impairment in Wistar rats.Methods: The study carried out in two groups of Wistar rats (n=6) with one group receiving DOX, and the other only normal saline used as control.During and after the experimentation period cognitive level of each rat was measured using Barnes maze (BM) till 8 weeks. At the end of theobservation period (after 8th week), a series of biochemical and histopathological studies were carried out after sacrificing the animals.Results: Errors to reach target and time taken to reach the target was found in BM experiment, elevated levels of antioxidants and hemoglobin wasfound reduced significantly in the DOX treated group when compared to the controls while histopathology of brain cells of DOX treated group alsoshowed reduction in hippocampal cellularity and cell death in hippocampal area suggesting evidence of oxidative damage caused by DOX treatment.Conclusion: Parameters in this study which not only conclusively show the damage caused to brain by DOX, but also estimates the changes caused toeach indicator by this drug.Keywords: Chemotherapy, Brain, Hippocampus, Oxidative stress

Pharmacognostic evaluation of Achyranthes coynei: Leaf

AbstractAchyranthes coynei Sant., (Amaranthaceae) is a rare, medicinal shrub, endemic to India. The plant especially, leaves are used in treatment of various disorders by folk healers. It has been scientifically evaluated for its antimicrobial and antioxidant properties. Pharmacognostic studies have not been carried out so far in this plant. So, the present study was undertaken to evaluate pharmacognostic characters of leaf of A. coynei. The studies were carried out in terms of morphological, microscopic characters and physicochemical parameters using standard methods. Leaf size (15–23 × 7–10 cm) and petiole nature (pink above and green beneath) were the distinguishing morphological character observed in the present study. Transverse section of leaf and petiole showed presence of long, multi-cellular, warty trichomes with pointed apex and short, glandular trichomes with blunt apex on thick walled epidermis; conjoint, collateral closed, endarch vascular bundles; rosette crystals of calcium oxalate in ground tissue. Ash and extractive values, micro and macro elements and nutritive factors were estimated. Leaf powder showed fluorescence under the influence of different solvents. Preliminary phytochemical screening showed the presence of alkaloids, saponins and triterpenoids. High Performance Thin Layer Chromatorgraphic (HPTLC) analysis yielded eight bands in leaf extract. The study forms the first report on pharamcognostic characters for A. coynei, which could be useful for identification and authentication of the plant

AN EXPERIMENTAL STUDY OF VRIKSHAYURVEDA SEED TREATMENTS ON GERMINATION RATE AND ACTIVE INGREDIENT OF BAKUCHI (PSORALEA CORYLIFOLIA LINN.) BY HPLC METHOD

Bakuchi (Psoralea corylifolia Linn.) is one of the important endangered medicinal plants used in Ayurveda and other Traditional systems. Its cultivation and propagation is difficult due to its low germination rate (5-7%) & prolonged seed dormancy. Bakuchi seeds made into 5 groups, the experiment was conducted in a complete randomized block design with 5 treatments and 5 replications totally 500 seeds in each group) & observed for 50 days. Control Group 1 no- seed treatment, Group 2- Standard treated with 1% conc H2SO4, Group 3 Vrikshayurvedic treatment done by soaking in milk subsequently fumigation of Vidanga & Ghee, Group 4- treated with paste of Brihati, Tila, Kamalanaala, Ghee & Group 5 treated by soaking in milk subsequently Cow dung, Vidanga & Honey applied. Number of seeds germinated, germination percentage, emergence index and relative seed germination parameters were observed. HPLC studies carried out of post harvested Bakuchi seeds of all 5 groups to know the effect of seed treatments on Psoralen content quantitatively. Overall results indicates that Group 4 (8.000 ± 0.8367) seeds soaked in 12 hrs milk followed by application of Brihati, Tila, Kamalanaala & Ghee paste for 12 hrs treatment is statistically significant (P value>0.05) in comparison with group 2 (4.600 ± 0.6782) Sulphuric acid treatment and Group 3 (4.200± 0.9165) fumigation with Honey & Vidanga. Rest of the groups shown insignificant changes on germination parameters. HPLC Results found that generally seed treatments may reduce the content of Psoralen as in control (Group 1) maximum percentage (0.04%w/w) of Psoralen is noticed. Among treatment groups Group 4 contains maximum (0.027%w/w) Psoralen next to control (0.039%w/w). Psoralen content is very less in standard Group 2 (0.022%w/w), Group 3 (0.023%w/w) & Group 4 (0.024%w/w). Maximum germination percentage was observed in Group 4 in comparison with the Group 2conventional method of treating with sulphuric acid. Estimation of Psoralen contents in the seeds from the plants grown by various treated seeds reveled that Group 4 is qualitatively better than standard, but inferior to the control, standard & other Vrikshayurveda seed treatment techniques used in the current experiment

Comparative HPTLC analysis of stem and leaf of Achyranthes coynei with Achyranthes aspera

Leaf and stem materials of Achyranthes coynei and Achyranthes aspera were used for HPTLC analysis. HPTLC plates were developed on n-hexane: ethyl acetate (5:1 v/v) solvent system. The densitometric profiles were evaluated to elucidate differences within and among the species. The Rf values and number of peaks obtained in densitrogram indicated chemical variation in the species. Although, both species had more or less equal number of peaks, their Rf values, %height and %area varied. Thus HPTLC analysis in absence of external standards, proved to be an informative tool for evaluating differences between these species and their parts

Elucidating type 2 diabetes mellitus risk factor by promoting lipid metabolism with gymnemagenin: An in vitro and in silico approach

Introduction: Adipose tissue functions as a key endocrine organ which releases multiple bioactive substances and regulate obesity-linked complications. Dysregulation of adipocyte differentiation, triglyceride metabolism, adipokines production and lipid transport contributes to impaired lipid metabolism resulting in obesity, insulin resistance and type 2 diabetes. Gymnema sylvestre plant is frequently used in Ayurveda for treatment of diabetes and obesity. Gymnemagenin is a major bioactive compound of Gymnema sylvestre. The present study was undertaken to elucidate the role of gymnemagenin in lipid metabolism by in vitro and computational approaches.Methods: A panel of twelve genes viz., Fasn, Lipe, Lpl, Pparg, Plin2, Cidea, Scd1, Adipoq, Lep, Ccl2, Fabp4, and Slc2a4, essential in lipid metabolism were selected and gene expression pattern and triglyceride content were checked in adipocytes (3T3L1 cells) with/without treatment of gymnemagenin by Real time PCR and colorimetric estimation, respectively. Mode of action of gymnemagenin on Pparg and Fabp4 was accomplished by computational studies. Gene set enrichment and network pharmacology were performed by STRING and Cytoscape. Molecular docking was performed by AutoDock vina by POAP pipeline. Molecular dynamics, MM-PBSA were done by Gromacs tool.Results:In vitro study showed that gymnemagenin improved triglyceride metabolism by up regulating the expression of lipase genes viz., Lipe and Lpl which hydrolyse triglyceride. Gymnemagenin also up regulated the expression of anti-inflammatory gene Adipoq. Importantly, gymnemagenin treatment up regulated the expression of Pparg gene and the downstream target genes (Plin2, Cidea, and Scd1) which are associated with adipogenesis. However, gymnemagenin has no effect on expression of Fabp4, codes for a lipid transport protein. In silico study revealed that gymnemagenin targeted 12 genes were modulating 6 molecular pathways involved in diabetes and obesity. Molecular docking and dynamics revealed that gymnemagenin stably bind to active site residue of Pparg and failed to bind to Fabp4 active site compared to its standard molecules throughout 100 ns MD production run. Gymnemagenin scored binding free energy of −177.94 and −25.406 kJ/mol with Pparg and Fabp4, respectively.Conclusion: Gymnemagenin improved lipid metabolism by increasing triglyceride hydrolysis (lipolysis), up regulating the crucial gene of adipogenesis and increasing the expression of anti-inflammatory adipokine proving its therapeutic importance as anti-obesity and anti-diabetic phytocompound

Ethyl gallate isolated from phenol-enriched fraction of Caesalpinia mimosoides Lam. Promotes cutaneous wound healing: a scientific validation through bioassay-guided fractionation

The tender shoots of Caesalpinia mimosoides Lam. are used ethnomedically by the traditional healers of Uttara Kannada district, Karnataka (India) for the treatment of wounds. The current study was aimed at exploring phenol-enriched fraction (PEF) of crude ethanol extract of tender shoots to isolate and characterize the most active bio-constituent through bioassay-guided fractionation procedure. The successive fractionation and sub-fractionation of PEF, followed by in vitro scratch wound, antimicrobial, and antioxidant activities, yielded a highly active natural antioxidant compound ethyl gallate (EG). In vitro wound healing potentiality of EG was evidenced by a significantly higher percentage of cell migration in L929 fibroblast cells (97.98 ± 0.46% at 3.81 μg/ml concentration) compared to a positive control group (98.44 ± 0.36%) at the 48th hour of incubation. A significantly higher rate of wound contraction (98.72 ± 0.41%), an elevated tensile strength of the incised wound (1,154.60 ± 1.42 g/mm2), and increased quantity of connective tissue elements were observed in the granulation tissues of the 1% EG ointment treated animal group on the 15th post-wounding day. The accelerated wound healing activity of 1% EG was also exhibited by histopathological examinations through Hematoxylin and Eosin, Masson’s trichome, and Toluidine blue-stained sections. Significant up-regulation of enzymatic and non-enzymatic antioxidant contents (reduced glutathione, superoxide dismutase, and catalase) and down-regulation of oxidative stress marker (lipid peroxidation) clearly indicates the effective granular antioxidant activity of 1% EG in preventing oxidative damage to the skin tissues. Further, in vitro antimicrobial and antioxidant activities of EG supports the positive correlation with its enhanced wound-healing activity. Moreover, molecular docking and dynamics for 100 ns revealed the stable binding of EG with cyclooxygenase-2 (−6.2 kcal/mol) and matrix metalloproteinase-9 (−4.6 kcal/mol) and unstable binding with tumor necrosis factor-α (−7.2 kcal/mol), suggesting the potential applicability of EG in inflammation and wound treatment

Ethnoveterinary practices in Uttara Kannada district of Karnataka

253-258Traditional methods of veterinary treatments using plants are predominant in rural folk of Uttara Kannada district situated in western Ghats of Karnataka. A total of 25 formulations from 39 plant species belonging to 30 families used to treat 21 diseases of domestic animals are described. The method of preparation, dose and duration of each plant along with its botanical name, family and local names are discussed

Effect of method and time of extraction on total phenolic content in comparison with antioxidant activities in different parts of Achyranthes aspera

The main purpose of this study was to evaluate the effect of extraction method with respect to time of exposure on total phenolic content and antioxidant potential of methanolic extracts (95%) from Achyranthes aspera leaves, stem and roots. Total phenolic content (TPC) was determined using the Folin–Ciocalteu method and antioxidant potential was tested using DPPH radical scavenging and FRAP assays. Plant populations aged 60 and 120 days were considered during the study. Overall, highest TPC was evident in mature plants (120 days) compared to younger (60 days). Similarly, leaves accumulated higher phenolics followed by roots and stem. Results showed the MAE technique to be efficient over USE and CSE methods. Same was evident in both the antioxidant assays tested. A similar trend was observed in both antioxidant assays as that of TPC, indicating phenolics to be the major contributor in the antioxidant potential of the plant. In conclusion it can be said that the yield of phenolic compounds depends on parameters viz. age of plant, part used for extraction, method of extraction and time required for the same

Preliminary pharmacognostic screening of Achyranthes coynei stem

Achyranthes coynei is a rare, endemic perennial shrub reported from Karnataka and Maharashtra states of India. The plant is used to treat various disorders by folk healers and was proven to have antimicrobial and antioxidant properties. The present study was undertaken to evaluate microscopic and macroscopic characters of A. coynei stem, along with its physicochemical parameters. ProgRes ® CapturePro and Microsoft Excel were used for statistical analysis. Perennial, shrubby nature and woody stem were the distinguishing morphological characters observed. Transverse section (TS) illustrated quadrangular outline of the stem and showed the presence of two types of trichomes on the thick-walled epidermis. TS also showed number of rosette calcium oxalates crystals; prismatic and microsphenoid crystals; conjoint, collateral open secondary vascular bundles; and two amphixylic medullary bundles in the pith. Ash and extractive values, micro and macro elements and nutritive factors were estimated in the present study. The presence of alkaloids, saponins and triterpenoids were observed in preliminary phytochemical screening. High-performance thin layer chromatographic analysis yielded different bands and also indicated the presence of oleanolic acid. The studied parameters for A. coynei stem will be useful for identification and authentication of the plant material