1 2717-204X Institute of Medicinal Plants, ACECR 645 Biotechnology Overview on Tissue Culture, Regeneration and Gene Transfornation to Artemisia annua L. Sharafi A b Hashemi Sohi H c Kazemi Tabar K d b National Institute for Genetic Engineering and Biotechnology, Tehran, Iran c National Institute for Genetic Engineering and Biotechnology, Tehran, Iran d Biotechnology Group, Mazandaran University, Mazandaran, Iran 1 12 2006 5 20 1 10 03 09 2006 16 12 2006 Artemisia annua L. a highly aromatic annual herb is a widely dispersed through out the temperate region containing the main active anti malaria compound artemisinin a sesquiterpene lactone. Because of low artemisinin level, unsuccessful chemical synthesis and slightly improvement of molecular breeding methods, biosynthesis of this valuable compound was noticed by using the tissue culture and Genetic Engineering in these years. The rate transformation could be varies with different factors like genotype, age, explant type and agrobacterium strain. The results recently was shown that explants in tissue cultures are very sensitive to kanamycin concentration and even it could be late shoot and root forrnation. Hairy root culture is another subject that is in these years was niticed as a alternative method by designing the new bioreactor for production if this secondrary metabolite.
646 Pharmacology & Toxicology Quantittative Analysis of Amount and Activity of Ficin in Khorasan’s Fig Tree Latex Retrieved from Different Organs of Tree in Different Seasons Rajabi O e Danaie Baghaki J f Varasateh AR g Jahangiri L h Baratian A i e School of Pharmacy, Mashhad University of Medical Sciences and immunology Research center, BooAli Research Comolex, Mashhad, Iran f School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran g School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran h School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran i School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran 1 12 2006 5 20 11 20 22 10 2005 04 12 2006 Background: The fig tree latex has been identified as a rich resource of "ficin" an enzyme protein. This enzyme has a vast application in food substances, pharmaceutical and clinical diagnosis, Fig plant can be harvested well in Khorasan province. Objective: To determine optimum organ and season to retrieve latex for different purposes. Method: In this study the measuring of the enzyme activity was based on its effect of casein substrate. Enzymatic activity was compared with standard papain solution activity. In this research calcium ion concentration as an enzyme activator was studied. Findings: The results show that the activity of the Khorasan native fig latex was 50-100 fold more than papain. The amount and the activity of ficin have been variant in different organs of the tree, and also varied with season. Calcium ion concentrations are also variable in different organs and are also season dependent. There is a relationship between calcium ion concentrations and specific activity of ficin. Results: The optimum season for latex collection is fall and the best organ is the branch ends. 650 Pharmacognosy & Pharmaceutics Evaluation of Antimicrobial Effects of 4 Medicinal Plants Against Salmonella typhymurium and Comparision them with Common Antibiotics in Veterinary Medicine Dakhili M j Zahraei Salehi T k Torabi Goodarzi M l Khavari A m j Assistant Professor of Microbiology and Immunology Department, Faculty of Medicine, Islamic Azad University of Qom, Iran k Associate Professor of Microbiology and Immunology Department, Faculty of Veterinary Medicine, University of Tehran, Iran l Departments of Veterinary Medicine, Agriculture and Natural Resources Research Center of Qom, Iran m Departments of Veterinary Medicine, Agriculture and Natural Resources Research Center of Qom, Iran 1 12 2006 5 20 21 26 13 03 2005 25 11 2006 Background: Thyme (Zataria multiflora Bois.), Penny voyel (Mentha pulegium L.) Origanum (Origanum vulgare L.) and Fennel (Foeniculum vulgare Miller) are the Iranian traditional spice which are widely used in the country. Following the recent efforts to look for healthy remedies with antimicrobial potential, the effects of essential oils of these medicinal plants were investigated on Salmonella typhimurum. Objective: Chemical composition and antimicrobial effects of essential oils from plants were evaluated on Salmonella typhimurum and compared with five antibiotics enrofloxacin, chloramphenicle, erytromycin, oxytetracyclin and flumequin. Methods: The essential oils of the leavs and stems are obtained from each plants by hydrodistillation and clevenger and analyzed by GC/MS. Results: The results of this study show that Thyme oil, compared to flumequin, oxytetracyclin and erythromycin is more effective (P<0.5). Essential oil of Penny voyel is more effective than oxytetracyclin and erythromycin. Among essential oil of plants, Thymol oil with minimal inhibitory concentration) MIC) 156.5µg/ml and minimal bactericidal concentration (MBC) 312.5 µg/ml had the best result against Salmonella typhimurum. Chemicial compounds: Thymol, Carvacrol and α -terpinen were found in Z. multiflora, M. pulegium, O. vulgar and F. vulgare by GC analyse. Conclusion: Considering the significant antibacterial effect of essential oil of Z. multiflora on effect on Salmonella typhimurum in invitro, it is recommened that, the therapeutic effect of this essential oil be studied in invivo. 651 Pharmacology & Toxicology Effect of Rosmarinus officinalis L. Aerial Parts Extract and Fractions on Morphine Withdrawal Syndrome in Mice Hosseinzadeh H n Ramezani M o Shahsavand Sh p n Corresponding author: Pharmacology and Toxicology Department, Pharmaceutical Research Center, Faculty of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran o Pharmacognosy and Biotechnology Department, Pharmaceutical Research Center, Faculty of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran p Pharmacist 1 12 2006 5 20 27 35 28 06 2004 16 10 2005 Background: Our previous studies have shown that the aqueous and ethanolic extracts of Rosmarinus officinalis L. aerial parts reduce morphine withdrawal syndrome in mice. Objective: As R. officinalis L. aerial parts total extracts reduced morphine withdrawal syndrome, we decided to evaluate the effects of rosemary extract fractions on the withdrawal syndrome. Method: Dependence was induced using subcutaneous injections of morphine daily for three days (50, 50 and 75 mg/kg). On the fourth day, morphine was injected two hours prior to the intraperitoneal injection of naloxone. The number of jumps during the 30 minute period after naloxone injection was considered as measure of the withdrawal syndrome. Results: The results indicate that the aqueous, methanolic-aqueous and chlorformic fractions (0.96 g/kg and 1.68 g/kg, i.p.) and also clonidine (0.3 mg/kg) and diazepam (5 mg/kg) reduce the number of jumps when they are injecte 1 h prior the last dose of morphine. Two evaluable MPLC chloroformic fractions also reduce the number of jumping in mice. In the open field activity, test, on the contrary of fraction 2, the fraction 1 and clonidine reduce locomotor activity. Conclusion: It is concluded that the reduction of withdrawal morphine syndrome by R. officinalis aerial parts fraction 1 may be due to the reduction of movement by this fraction, but the fraction 2 may reduce withdrawal syndrome via an interaction with opioid system. 652 Pharmacognosy & Pharmaceutics The Study of Quantitative and Qualitative Changes of Essential Oil from Smyrnium cordifolium Boiss. in Lorestan Province Esmaili A Amiri H Department of Chemistry, Tehran North Campus, I. A. University, Tehran, Iran Biology Department, Lorestan University, Khoramabad, Iran 1 12 2006 5 20 36 41 27 11 2004 20 12 2006 Background: Smyrnium cordifolium belongs to Umbelliferae family which growing in the west and southwest of Iran and has nutritive and medicine use in the regions. Aim: Identification of chemical compositions of essential oil of S. cordifolium grown in different regions of Lorestan province and study effects of environmental factors on it. Methods: This plant was collected from five differen regions of Lorestan province and was investigated for quatitative and qualitative changes of its essential oil. The oils was injected to GC for identification of constituents. Results: Yields of essential oil were 0.07% to 0.55% in these different regions. The most amount of essential oil was related to Poldokhtar and the least amount to Aligoodarz. In the essential oil of different regions two oxygenated sesquiterpen Curzerene and Curzerenone had the most amount. Among common and remarkable components we can mention to Germacrene-D, Isopimarol and Phyllocladanol. In Kouhdasht and Khoramabad, Curzerene was the main component with 39.4 and 34.2 percentages respectively, and in Poldokhtar, Nourabad and Aligodarz it was Curzerenone with 49.4, 33.8 and 35.6 percentages.Also, in different regions Germacrene-D is abundant in the oil Discussion: The identified components of essential oil of different regions showed the differences and similarities from quantitative and qualitative point of view. GC/MS analysis revealed that the sesquiterpen compounds were the main components in different regions. α-pinene and β-pinene were just observed in Norabad and Aligodarz. 653 Pharmacognosy & Pharmaceutics The Evaluation of Eucaliptus camaldolensis Dehnh. Esscence Application in Control of Fungal Pollution of Trout Eggs Ebrahimzadeh Mousavi H Rohani MS Khowsravi AR Mehrabi Y Basti A Associated Prof. of Vet Faculty, Tehran University, Tehran, Iran Scientific board of Iranian Fisheries Research Organisation (IFRO) Proffessor of Vet Faculty, University of Tehran, Tehran, Iran Expert of Iranian Fisheries Organisation Associated Prof. of Vet Faculty, Tehran univ. Tehran, Iran 1 12 2006 5 20 42 47 17 11 2004 12 12 2006 Background: The use of medical plants is necessary as the antifungal agent substitute the malachite green for control of fungal pollution in salmonid hatcharies. Objective: The capability of Eucaliptus camaldolensis escence in control of fungal pollution about 50 kg trout eggs 4 years old Oncorhynchus mykiss brooders were evaluated in clinical and farm conditions, with using of 1,5,10,25,50 and 100 ppm dosages of this escence daily with 3 repetition until the eyed egg stage (about 18-20 days). Besides, under the equal condition, the positive control group was treated with 1 ppm malachite green and the negative control group was assessed without any drugs. Results: After culturing of fungal eggs, a pollution of Saprolegnia parasitica and Fusarium solani was obsereved. MIC result of Eucaliptus escence against Saprolegnia was 2.3 and aginst Fusarium was 8.2 ppm. Measuring the physical and chemical factors of water, no negative effect is recognized. Conclusion: The eucalyptus escence specially in the doses of 25, 50 and 100 ppm is significantly different from negative control group (p< 0.05) and created suitable inhibition aginst water fungi. 654 Pharmacognosy & Pharmaceutics Inhibit Effect of Allium hirtifolium Boiss. (Persian shallot) Hydroalcoholic Extract on the Growth of Leishmania infantum in vitro Amanzadeh Y Izaddoost M Soltanpoor A Mahami M Taheri M Khalifeh Gholi kalani M Kalantari N Taran M Sadat Ebrahimi SE Department of Pharmacognosy, College of pharmacy, Tehran University of Medical Sciences, Tehran, Iran Department of Pharmacognosy, College of pharmacy, Tehran University of Medical Sciences, Tehran, Iran Department of Pharmacognocy, College of pharmacy, Tehran University of Medical Sciences, Tehran, Iran School of Public Health, Institute of Public Health Research, Tehran University of Medical Sciences, Tehran, Iran College of Medicine, Tehran University of Medical Sciences, Tehran, Iran School of Public Health, Institute of Public Health Research, Tehran University of Medical Sciences, Tehran, Iran College of Medicine, Tehran Univesity of Medicinal Sciences, Tehran, Iran School of Public Health, Institute of Public Health Research, Tehran University of Medical Sciences, Tehran, Iran Department of Pharmacognosy, College of pharmacy, Tehran University of Medical Sciences, Tehran, Iran 1 12 2006 5 20 48 52 24 01 2005 22 11 2006 Background: Antiprotozoal effects of genus Allium plants including garlic and onion have been proven in Aryana Lands since hundreds years ago. Objective: In this study, inhibitory effect of persian shallot hydroalcoholic extract on growth of Leishmania infantum was evaluated in vitro. Methods: Different concentrations of the extract including 0.01, 0.05, 0.1 and 0.2 mg/cc in schneider medium were prepared, the parasites were added and counted every day during 7 days after culture. Results: Concentrations between 0.01 - 0.1 mg/cc inhibited the growth of parasites on the third day, but the highest concentration (0.2 mg/cc) inhibited the growth of parasites on the first day. Conclusion: This study proves inhibitory effect of persian shallot hydroalcoholic extract (containing allicin, ajoene and other agents) on Leishmania infantum. For exact evaluation of persian shallot antileishmanian properties, it is necessary to evaluate inhibitory effect of the plant hydroalcoholic extract in vivo. 655 Traditional Pharmacy & Traditional Medicine Study of the ACE Inhibitory Effect of Medicinal Plants Used in Iranian Folk-Medicine as Antihypertensive Remedy Ziai SA Rezazadeh Sh Dastpak A Shabestari A Taghizadeh M Naghdibadi HA Poorhoseini L Department of Pharmacology, Institute of Medicinal Plants, ACECR, Tehran, Iran Institute of Medicinal Plants, ACECR, Tehran, Iran Department of Pharmacology, Institute of Medicinal Plants, ACECR, Tehran, Iran Pharmacist, Faculty of Pharmacy, Islamic Azad University, Tehran, Iran Institute of Medicinal Plants, ACECR, Tehran, Iran Institue of medicinal plants, Iranian Academic Center for Education, Culture, and Research (ACECR) & Tarbiat Modares University Tehran, Iran Department of Pharmacology, Institute of Medicinal Plants, ACECR, Tehran, Iran 1 12 2006 5 20 53 74 04 09 2006 20 11 2006 Background: Angiotensin converting enzyme (ACE) is an exopeptidase which converts angiotensin I to angiotensin II. Angiotensin II causes vasoconstriction and aldosteron secretion and inactivate bradykinin, so ACE has major role on blood pressure regulation. Objective: In this study 135 plants used in Persian traditional medicine have been investigated for their angiotensin converting enzyme (ACE) inhibitory activity potential. They were selected on the basis of their usage as antihypertensive, cardiotonics & diuretics. Methods: The dried powdered plant material, 1 g was extracted with 10 ml water and ethanol (%96) in ultrasonic bath. The extracts were filtered and concentrated in vacuum except for the water extracts, which were freeze-dried. Test solutions were made by dissolving extract in assay buffer, corresponding to a final concentration of 0.33 mg of crude plant extract in 1 ml test volume. Enzyme assay was performed by HPLC method. Plants exhibiting inhibition levels more then 50% were further tested for the presence of tannins in order to eliminate possible false positives. Results: In total, 52 species out of the 135 (39%) screened, gave more than 50% ACE inhibition. 40 species were found to possess a high ACE inhibiting ability and were low in their tannin content. 656 Pharmacology & Toxicology The Clinical Investigation of Securigera securidaca (L.) (Degen & Doerfler) Seeds in Type II Diabetic Patients a Randomized, Double-Blind, Placebo-Controlled Study Fallah Huseini H Hooseini P Heshmat R Yazdani D Rahmani M Hemati Moqadam HR Larijani B Alavi SHR Department of Pharmacology, Institute of Medicinal Plants, ACECR, Tehran, Iran Endocrinology & Metabolism Research Center, Tehran University of Medical Sciences Tehran Iran Department of Epidemiology and Biostatistics, Tehran University of Medical Sciences. Tehran, Iran Department of Pharmacology, Institute of Medicinal Plants, ACECR, Tehran, Iran Endocrinology & Metabolism Research Center, Tehran University of Medical Sciences Tehran Iran Department of Pharmacology, Institute of Medicinal Plants, ACECR, Tehran, Iran Endocrinology & Metabolism Research Center, Tehran University of Medical Sciences Tehran Iran Department of Pharmacognosy, Faculty of Pharmacy and Medicinal Plant Research Center, Terhan University of Medical Sciences, Tehran, Iran 1 12 2006 5 20 75 79 04 09 2006 11 11 2006 Backgrounds and Aims: Securigera securidaca (L.) seeds are used by traditional herbalists for the treatment of diabetes in several parts of Iran. Clinical investigation of its efficacy and its toxicity in diabetic patients is of importance. Methods: A 2-month randomized double blind clinical trial was conducted in 70 type II diabetic patients in two well – matched groups. One group (n=35) received 500 mg Securigera securidaca (L.) seeds capsule 3 times a day plus standard therapy, while the control group (n=35) received placebo plus standard therapy. The patients were visited monthly and glycosylated hemoglobin (HbA1c), fasting blood glucose, total cholesterol, LDL and HDL, triglyceride, SGOT, SGPT, ALK, BUN, serum creatinine levels were determined at the beginning and the end of the study after two months. Results: There was no significant change in HbA1c and fasting blood glucose level in Securigera securidaca (L.) seeds treated patients after two months as compared to beginning of the treatment and also as compare to placebo group. Serum lipid level also did not change significantly. The liver enzyme level and creatinine level in blood did not change significantly in Securigera securidaca (L.) seeds treated patients. No side effect was observed in any of patients in both of the groups. Conclusion: The results show that the Securigera securidaca (L.) seeds treatment with known antidiabetic property in traditional herbal therapy has no beneficial effect on improving glycemic profile in type II diabetic patients. The use of this herbal medicine in the dose of 1500mg in 3 divided dose showed no gasterointestinal symptom as well as liver and kidney abnormality during two months therapy. The long term efficacy and toxicity of this herbal medicine in higher dosage may be investigated in large multi center clinical study.