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KMITL SCIENCE AND TECHNOLOGY JOURNAL VOL.13, NO.1, 2013 Antimotility Effect of Machiluss odoratissima & Sonchus wightianus from Nepal Amit Subedi§‡*, Dipak Khakural§, Sadhana Amatya§, Tirtha Maiya Shrestha§§, §Department of Pharmacy, Institute of Medicine, Tribhuvan University, Kathmandu, Nepal. ‡ College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk 712-749, South Korea. §§ Department of Pharmacy, Kathmandu University, Nepal. † Department of Plant Resources, Kathmandu, Nepal. The extracts of Machiluss odoratissima & Sonchus wightianus were subjected to investigate phytochemical, cytotoxic and antimotility properties. The phytochemical Barks of M. odoratissima and leaves of S. screening showed the presence of different compounds. The wightianus were collected from Phulchowki and cytotoxic screening showed that both the extracts are non- Sanepa respectively of Kathmandu Valley during the toxic. The in vivo study showed the presence of potent month of November. Brine shrimp from San Francisco Bay (USA) was used and Swiss albino mouse were purchased from Department of plant resource, Keywords: Machiluss odoratissima, Sonchus wightianus,
M. odoratissima extract – Briefly, cleaned, air dried, and powdered barks were macerated by Diarrheal disease is responsible for deaths of methanol. The methanolic fraction was defatted by millions of children especially in developing countries using petroleum ether. Finally, the methanolic portion [1]. The commonly used antimotility drugs, was dried by vacuum evaporator at temperature loperamide and diphenoxylate, are not free from associated risk and have limited use in children. Thus, S. wightianus extract – Briefly, the cleaned, air for the search of new, safe and efficacious antimotility dried and powdered leaves were successively extracted in Soxhlet apparatus by petroleum ether, diethyl Plants have been the source of number of chemical ether, methanol and water. The extracts were dried identity with pharmacological values. Though many by keeping in water bath below 70°C. Methanolic plants in developing countries are used as traditional portion was used during all the experiments. medicine, the science behind their use is still unknown. Besides traditionally used medicinal plants, there are still many other plants with their medicinal values unknown and unexplored. Phytochemical screening of the extracts was done S. wightianus, locally called Dudhe, of asteraceae to identify the main groups of chemical constituents family is found in 700-4500 m in Nepal while M. by their color reactions as described elsewhere [2, 4]. odoratissima, locally called Kaulo, of lauraceae family is found in 2100-2300 m on lower part of Himalaya [2]. S wightianus is traditionally used during diarrhea Brine shrimp bioassay was done according to [3] but not yet evaluated scientifically. M. Carballo [5]. Briefly, artificial seawater was prepared odoratissima is not used traditionally for medicinal and brine shrimps were hatched in beaker by purpose but reported to have antioxidant and sparkling brine shrimp eggs at 1gm/ltr. It was antimicrobial property [2]. Thus, in our present study continuously illuminated with table lamp and we chose one traditionally used medicinal plant and temperature was maintained at 25-30°C. After 48 hrs, one with no traditional use to evaluate their ten highly active phototropic nauplii were transferred to test tube. Stock solutions of the extracts were * Corresponding author: prepared and required volume was transferred to test tube and final volume was adjusted to 5 ml by adding sea water. Dry yeast suspension was added in each KMITL-STJ: Research report/Finding, Part A: Material Science test tube as a source of food. After 24 hrs, the number Amit Subedi et. al.: Antimotility effect of Machiluss odoratissima & Sonchus wightianus from Nepal of dead nauplii in each test tube was counted against bright background. The percentage mortality of brine TABLE II. PHYTOCHEMICAL SCREENING OF DIFFERENT EXTRACTS OF shrimps was calculated at each concentration and M.ODORATISSIMA AND S.WIGHTIANUS. PE = PETROLEUM control. LC (lethal concentration for 50% ETHER, DE = DIETHYL ETHER, MET = METHANOL population) of the extracts weredetermined by regression equation obtained by plotting mean % mortality against logarithm of concentration. Results shown are average of five experiments for each dose. Charcoal meal test was used to evaluate the efficacy of a compound to inhibit gastrointestinal motility in small intestine as described by Rouf [6]. Ethical approval for animal experiments was taken from Institutional Review Board of Institute of Medicine, Kathmandu, Nepal. Briefly, mouse of 20-25 gm were starved for 20 hrs prior to experiments and were divided into 4 groups of 4 mice each. Control group received distilled water at 10 ml/kg, positive control group received atropine at 5 mg/kg and test group received extract intraperitoneally at specified dose prepared in volume of 10ml/kg. After 30 minutes, each group received 1 ml charcoal meal per oral (charcoal meal = 12 gm activated charcoal, 2 gm gum tragacanth diluted to 130 ml by distilled water). The brine shrimp bioassay showed that the LC After 30 minutes of charcoal meal, each mouse was for methanolic extract of M. odoratissima is 48046 killed. The abdomen was opened and intestine from µg/ml while that of S. wightianus is 2248.72 µg/ml as pylorus to ileocaecal junction was cut and its length shown in Table III. The LC values of both the measured. The distance which charcoal meal travelled extracts are very higher than nontoxic levels [7]. So it from pylorus was measured and expressed as is likely that the extract or the lead compound responsible for the pharmacological effect has very BRINE SHRIMP BIOASSAY OF DIFFERENT EXTRACTS OF Distance travelled by charcoal meal M.ODORATISSIMA AND S.WIGHTIANUS. 3.1 Extractive values and Phytochemical Screening The extracts were prepared as described and the extractive values are shown in Table I. The phytochemical screening showed that M. odoratissima contains terpenoids, tannins, deoxy sugar, saponin and phenolic compounds while S. wightianus contains glycoside, steroid, tannin, flavonoids, deoxy sugar and The extract of S.wightianus and M. odoratissima showed potent antimotility effect in Swiss albino mice EXTRACTIVE VALUES OF M.ODORATISSIMA AND S.WIGHTIANUS IN DIFFERENT SOLVENT SYSTEM as shown (Fig.1). The extract of S. wightianus was more potent than that of M. odoratissima. The present in vivo study showed that extracts of both the plant possess promising antimotility effect. The crude extract showed more potent action than atropine. Thus we can conclude that both the plants possess potent and safe antimotility compound. The extract of S. wightianus showed dose dependent activity. Surprisingly, the effect of extract of M. odoratissima was not dose dependent (Fig. 1). From this study we could not find out the reason but it is possible that the M. odoratissima extract contains promotility and antimotility compounds and KMITL SCIENCE AND TECHNOLOGY JOURNAL VOL.13, NO.1, 2013 the effect of antimotility compound got saturated at 50 mg/kg while the promotility compound still showing dose dependent effect at 50 mg/kg. We would like to give special thanks Purna Bahadur Rana and Bishal Raj Adhikari for their assistance and contribution during the study. [1] Ojewole JAO, Awe EO, Chiworroro WDH, 2008. Antidiarrhoeal activity of Psidium guajava Linn. (Myrtaceae) leaf aqueous extracts in rodents. J. Smoth Muscle Res., 44(6), 195-207 [2] Subedi A, Amatya MP, Shrestha TM, Mishra SK, Pokhrel BM, 2012. Antioxidant and antibacterial activity pharmacopoeia used in traditional system of medicine for the treatment of diarrhoea in Arunachal Pradesh. Medicinal Plants - International Journal of Phytomedicines and Related Figure 1. Antimotility effect of extracts of M.odoratissima
and S.wightianus in Swiss albino mice. Data are [4] Paudel A, Panthee S, Shakya S, Amatya S, mean ± S.E of 4 mice. *, statistically significant Shrestha TM, Amatya MP, 2011. Phytochemical (p ≤ 0.05) and indicates significance against both and antibacterial properties of Rhododendron campanulatum from Nepal. Journal of Traditional Medicines, 6(6), 252-258. [5] Carballo JL, Hernandez-Inda ZL, Perez P, Garcia- Gravalos M, 2002. A comparison between two brine The present study showed that both plants shrimp assays to detect in vitro cytotoxicity in marine contain compounds with significant antimotility natural products. BMC Biotechnology, 2(17). effect. Extracts from both plants can be used to [6] Rouf R, Uddin SJ, Shilpi JA, Alamgir M, 2007. develop new, potent and safe antimotility drug. This Assessment of anti diarrhoeal activity of the methanol study shows that scientific study also needs to be extract of Xylocarpus granatum bark in mice model. focused on plants without traditional medicinal values Journal of Ethnopharamcology, 109(3), 539-542. in the search of novel drug. Though we have not yet [7] Moshi MJ, Innocent E, Otieno JN, Magadula JJ, Nondo RSO, Otieno DF, Wensheit A, Mbabazi P, antimotility effect, detailed studies of both plants 2010. Antimicrobial and brine shrimp activity of involving isolation, purification and characterization Acanthus pubescens root extracts. TJHR, 12(2). pharmacological effects are going on and will be reported in future.


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