2012 Annual Report
1a.Objectives (from AD-416):
The long-term objective of this project is to identify and chemically characterize secondary metabolites from natural sources with nutraceutical and/or pesticidal properties as chemical markers for selection of high value crops and/or lead compounds for structure-activity optimization. Over the next 5 years, we will focus on the following objectives:
Objective 1. Profile phytochemicals of selected crops (blueberry and rice) to identify species or cultivars with high contents of compounds with beneficial health and/or fungicidal properties to promote development as high value crops.
Objective 2. Discover and identify technologies based on secondary metabolites that facilitate the development of commercially viable herbicides, fungicides, insecticides, insect repellants, fish bacterides, and health promoters.
1b.Approach (from AD-416):
Blueberry and rice samples will be extracted with appropriate solvents and the extracts will be analyzed by liquid chromatography-mass spectrometry, as well as by gas chromatography-mass spectrometry. The extracts will also be tested for antioxidant activity using a cell-based antioxidant assay. The chemical profiles of the extracts will be correlated with disease resistance of the samples, as well as antioxidant activity. A bioassay-guided investigative approach will be performed on bioactive extracts in efforts to discover herbicides, fungicides, insecticides, insect repellents, fish bactericides and health promoters. General procedures for isolation of active compounds such as solvent extractions, distillations, column chromatography and thin layer chromatography will be employed. Extracts, fractions and pure compounds will be tested for phytotoxicity, antifungal and insecticidal activity in assays being carried out routinely at NPURU and at collaborator's laboratories. Identification of active compounds will be done using spectroscopic methods (Ultraviolet (UV), Infrared (IR), Mass Spectroscopy (MS), Nuclear Magnetic Resonance (NMR)). Chemical studies modification will be performed on compound(s) that will be identified as "highly active" to improve activity or bioavailability.
Objective 1 of the project (Profile phytochemicals of selected crops to identify species or cultivars with high contents of compounds with beneficial health and/or fungicidal properties to promote development as high value crops) was met. Field-grown blueberry cultivars Duke, Spartan, Berkeley and Brigitta Blue were received from collaborator. Chemical profiling has been completed and data analysis is currently being undertaken. The antioxidant activity of the extracts is also currently being performed by a collaborator. Objective 2 of the research project (Discover and identify technologies based on secondary metabolites that facilitate the development of commercially viable herbicides, fungicides, insecticides, insect repellants, fish bacterides, and health promoters) was met. In continuing studies to discover health promoting properties of pterostilbene, it was shown to have anti-obesity effect in animals fed high-fat diet. Two doses of pterostilbene were used in the study: 15 and 30 mg/kg body weight; PT15 and PT30 group, respectively. Pterostilbene significantly reduced total adipose tissue mass (15.1% in PT15 group and 22.9% in PT30 group) without changes in food intake. Biochemical analyses showed pterostilbene inhibited the activities of key enzymes involved in lipid synthesis, in the live and adipose tissue. Additionally, the PT30 dose also increased fatty acid oxidation in the liver. Serum levels of pterostilbene were also determined, and correlated with results obtained from biochemical analyses. These new findings led to a patent application on the use of pterostilbene as anti-obesity agent. In a continuing study on breadfruit, Artocarpus altilis, to identify chemicals responsible for mosquito deterrence, capric, undecanoic, and lauric acid were identified as the primary components responsible for the activity. All three of these compounds were significantly more effective than DEET at repelling mosquitoes. During the past year scientists with the Natural Products Utilization Research Unit (NPURU) at Oxford, Mississippi, have extensively expanded international collaborations in China and Brazil to discover and evaluate natural products that would not normally be accessible to NPURU scientists. Research involvement in the Deployed War-Fighter Protection Program that began in 2007 continued to expand in 2012 as NPURU increased its output of extracts and pure compounds for evaluations for insecticidal and mosquito deterrent activity through collaborations with the Center for Medical, Agricultural, and Veterinary Entomology and an entomologist at the National Center for Natural Products Research. Phytochemical investigation was performed on the following plants: Angelica sinensis, Evodia rutaecarpa, and Pileostegia viburnoides to isolate and identify insecticidal and fungicidal compounds. In yet another study relevant to Objective 2, compounds were prepared by synthetic modification of the structure of the natural compound wogonin and were tested for inhibition of the fish bacteria Flavobacterium columnare. Some analogs showed good activity.
Breadfruit compounds as mosquito repellants. Mosquitoes are vectors of some infectious diseases. Discovery of repellants is warranted to replace DEET-based commercial repellants due to some untoward side effects of DEET. Breadfruit, Artocarpus altilis, is a tropical staple food crop traditionally cultivated throughout Oceania (Melanesia, Micronesia, and Polynesia). Dried male inflorescences of breadfruit are burned in communities throughout Oceania to repel flying insects, including mosquitoes. In a collaborative study by a scientist at NPURU in Oxford, MS, in collaboration with scientists at the University of British Columbia Okanagan and the National Tropical Botanical Garden, capric, undecanoic, and lauric acid were identified as the primary components responsible for the insect-deterrent activity of the burning breadfruit. All three non-toxic compounds were significantly more effective than DEET at repelling mosquitoes. This study suggests that the inflorescences could be developed into a valuable secondary product for local use or income generation.
Anti-obesity effect of pterostilbene. Obesity is a health problem spreading globally and contributes to major chronic diseases such as diabetes and heart disease. ARS researcher at the Natural Products Utilization Reserch Unit (NPURU) in Oxford, MS, and collaborator at the University of the Basque Country in Spain showed in animal studies that pterostilbene, a naturally-occurring phenolic compound, reduces total adipose tissue mass, without changes in the animals’ food intake. Biochemical analyses further showed pterostilbene inhibited the activities of key enzymes involved in lipid synthesis, in the live and adipose tissue, and also increased fatty acid breakdown in the liver. Serum levels of pterostilbene were in concordance with biochemical results. These new and significant findings led to a patent application. This study adds another health benefit of pterostilbene, for which USDA had patents on and that have been licensed. This study will boost the sales of pterostilbene products already on the market, including tier-one pharmacy stores. From agricultural standpoint, this study will help blueberry farmers since pterostilbene has been identified in blueberries. This study will help the general public in preventing or alleviating obesity and associated health problems.
Nakano, H., Cantrell, C.L., Mamonov, L.K., Osbrink, W.L., Ross, S. 2011. Echinopsacetylenes A and B, new thiophenes from Echinops transiliensis. Organic Letters. 13(23):6228-6231.
Kocyigit-Kaymakciogl, U., Oruc-Emre, E., Unsalan, S., Iscan, G., Demirci, F., Rollas, S., Tabanca, N., Wedge, D.E., Khan, S.I. 2011. Synthesis and biological activity of hydrazide hydrazones and their corresponding 3-acetyl-2,5-disubstituted-2,3-dihydro-1,3,4-oxadiazoles. Bioorganic and Medicinal Chemistry. 1-10.
Ozek, G., Ishmuratova, M., Tabanca, N., Radwan, M.M., Goger, F., Ozek, T., Wedge, D.E., Becnel, J.J., Cutler, S.J., Baser, K.H. 2012. One-step multiple component isolation from the oil of Crinitaria tatarica (Less.) Sojak. by preparative capillary GC with characterization by spectroscopic and spectrometric techniques and evaluation of biological activity. Journal of Separation Science. 35(5-6):650-660.
Cannon, J.B., Cantrell, C.L., Astatkie, T., Zheljazkov, V.D. 2012. Modification of yield and composition of essential oils by distillation time. Industrial Crops and Products. 41:214-220.
Cantrell, C.L., Ali, A., Duke, S.O., Khan, I. 2011. Identification of the mosquito biting deterrent constituents from the Indian folk remedy plant Jatropha curcas. Journal of Medical Entomology. 48(4):836-845.
Cantrell, C.L., Dayan, F.E., Duke, S.O. 2012. Natural products as sources for new pesticides. Journal of Natural Products. 75(6):1231-1242.
Dandawate, S., Williams, L., Joshee, N., Rimando, A.M., Mittal, S., Thakur, A., Lum, L., Parajuli, P. 2011. Scutellaria flavonoids inhibit tumor-mediated induction of Treg cells via inhibition of TGF-ß1 activity. Cancer Immunology and Immunotherapy. 61:701-711.
Chauhan, K.R., Jones, M., Klun, J.A., Cantrell, C.L., Ragone, D., Brown, P., Murch, S. 2012. Isolation and identification of mosquito (Aedes aegypti) biting deterrent fatty acids from male inflorescences of breadfruit (Artocarpus altilis (Parkinson)Fosberg). Journal of Agricultural and Food Chemistry. 60:3867-3873.
Parajuli, P., Joshee, N., Chinni, S.R., Rimando, A.M., Mittal, S., Sethi, S., Yadav, A.K. 2011. Delayed growth of glioma by Scutellaria flavonoids involve inhibition of Akt, GSK-3 and NF-kB signaling. Journal of Neuro-Oncology. 101:15-24.
Cantrell, C.L., Klun, J.A. 2011. Callicarpenal and Intermedeol: Two Natural Arthropod Feeding Deterrent and Repellent Compounds Identified from the Southern Folk Remedy Plant, Callicarpa americana. In: Paluch, G.E., Coats, J.R., editors. Recent Developments in Invertebrate Repellents. Volume 1090. Washington, DC: American Chemical Society. p. 47-58.
Chang, J., Rimando, A.M., Pallas, M., Camins, A., Porquet, D., Reeves, J., Shukitt Hale, B., Smith, M.A., Joseph, J.A., Casadesus, G. 2011. Low-dose pterostilbene but not resveratrol is a potent neuromodulator in aging and Alzheimer’s Disease. Neurobiology of Aging. 33:2062-2071.
Queiroz, S.C., Cantrell, C.L., Duke, S.O., Wedge, D.E., Nandula, V.K., Moraes, R.M., Cerdeira, A.L. 2012. Bioassay-directed isolation and identification of phytotoxic and fungitoxic acetylenes from Conyza canadensis. Journal of Agricultural and Food Chemistry. 60:5893-5898.
Mikstacka, R., Rimando, A.M., Dutkiewicz, Z., Stefanski, T., Sobiak, S. 2012. Design synthesis and evaluation of the inhibitory selectivity of novel trans-resveratrol analogues on human recombinant CYP1A1 CYP1A2 and CYP1B1. Bioorganic and Medicinal Chemistry. 20(17):5117-2126.
Zhang, A., Rimando, A.M., Fish, W., Mentreddy, S.R., Mathews, S. 2012. Serviceberry [Amerlanchier alnifolia (Nutt.) Nutt. ex. M. Roem(Rosaceae)] leaf exhibits mammalian alpha glucosidase activity and suppresses postprandial glycemic response in a mouse model of diet induced obesity/hyperglycemia. Journal of Ethnopharmacology. 143(2):481-487.
Poulose, S.M., Fisher, D.R., Larson, J., Bielinski, D.F., Rimando, A.M., Carey, A.N., Schauss, A.G., Shukitt Hale, B. 2012. Anthocyanin-rich acai (Euterpe oleracea mart.) fruit pulp fractions attenuate inflammatory stress signaling in mouse brain BV-2 microglial cells. Journal of Agricultural and Food Chemistry. 60:1084-1093.