Progress 09/01/08 to 08/31/11
Outputs
OUTPUTS: Propagation: a successful protocol for stem cutting propagation of V. uliginosum was developed and permanent field plots of V. uliginosum and V. vitis-idaea were established at the Georgeson Botanical Garden for use in antioxidant, genetic, biochemical diversity research. The garden is visited by more than 20,000 people annually, and interpretive signs explain the research. Research results are shared with individual berry growers and their help has been enlisted in collecting large-fruited germplasm from wild stands. Controlled environment research continues on light and environmental conditions best suited for crop establishment, early season development, humidity control, flowering and fruit set, plant morphological attributes, frost protection, yields and nutritional berry quality. Chemistry developed bioassays that identified both antioxidant and anti-inflammatory properties of blueberry extracts and isolated fractions using the TAP assay and sphingomyelinase assay. They developed an improved method for isolating blueberric acid by running an extract rich in blueberric acid through an ion exchange column (weakly basic tertiary amine) using deionized water as an eluant until all the non ionic components (mainly fructose) have eluted. Research continues to explore a method to remove the small amount of remaining malic acid impurity from the blueberric acid by precipitating malic acid as its 2,4-dinitrophenylhdrazone derivative. Through other funding work continues in basic neuroscience targeted toward the neutraceutical market and concentrating on specific characteristics of Alaska berries and their affects on neurological responses. Outreach: Cooperative Extension Service reports that twenty five berry products are being produced by small businesses with six more products in the planning stages. Products are wildberry sauces to be served with desserts, baked goods, ice cream, jams and jellies, and candies. PARTICIPANTS: Carol E. Lewis, Director AFES, Project Oversight. PROPAGATION: Patricia Holloway, Professor of Horticulture, Research Technician Grant Matheke. Two graduate students, Tina Buxbaum, and Rehanon Pampell, working on pollination biology and bumble bee biology to enhance fruit set in wild and cultivated stands using native pollinators. COLLABORATORS: Alaska Berry Growers Association, Kenai, commercial berry producer, Brian Olsen, Kenai, growers in Prince of Wales Island, two growers in the Interior, Charles Knight and Papa Meneur, and collaborative team USDA ARS: entomologist, Dr. Alberto Pantoja, UAF entomologist, Dr. Derek Sikes, and Alaska beekeeper, Stephen Peterson to expand knowledge of native pollinators. CONTROLLED ENVIRON: Meriam Karlsson, Professor of Horticulture, project manager; Jeff Werner, Research Professional; Jeffrey Johnson, Research Associate; Graduate Student: Yosuke Okada. Undergraduate students: James Ward, Andrew Winkelman, Patrick Sanders. COLLABORATORS:Chena Hot Springs Resort, Collaborator, Pike's Waterfront Lodge, Collaborator, Rough Brothers Greenhouse Manufacturing Company (Cincinnati, Ohio), Collaborator, Link4 Corporation (Placentia, California), Collaborator, Svensson Screens (Charlotte, North Carolina), CHEMICAL ANALYSIS: Thomas Kuhn Associate Professor, in close collaboration with Thomas Clausen Department Chair and Professor of Chemistry and Biochemistry and Lawrence Duffy, Professor of Chemistry and Biochemistry. Fractionation of AK wild bog blueberries was accomplished and driven by his graduate student Colin McGill, who graduated recently with a Ph.D degree in biochemistry based on his investigations .Sally Gustafson, PhD graduate student, succeeded in demonstrating the capacity of blueberries and purified compounds to potently inhibit NOX. Dr. Kriya Dunlap has accomplished the fractionation of an nSMase inhibitor from AK wild bog blueberries in collaboration with Dr. Colin McGill. Her efforts were recognized by the Dean of the College of Natural Sciences and Mathematics. She was promoted to a Research Faculty at UAF providing her status as an independent principal investigator. Sigourney Walker (Undergraduate student) participated in many of the isolations including IV, fructose, and cyaniding. Talal Ahmed, Gavin Baker, Patrick Bollig, Jennifer Chambers, Carrie Duplisea, Christopher Ebner, Kathleen Gerdes, Andy Lester and Brad Weis (Organic Laboratory {Chem 324W} students) performed the first two steps in the synthesis described. COLLABORATORS: Dr. James Joseph, USDA Human Nutrition Research Center at Tufts University, Dr. Donald Ingram, Pennington Biomedical Research Center (formerly NIA), and Dr. Wilhelmina Kalt,Agriculture Canada. A new collaboration was established with Dr James Bamburg, Colorado State University. The goal of the interaction between the two laboratories is to investigate whether AK wild bog blueberries, fractions derived, and purified compounds harbor the potency to rescue neurons from a reorganization of their actin cytoskeleton into rod-like structures. Bamburg and co-workers demonstrated the formation of rods as an early event in neurodegeneration. TARGET AUDIENCES: Blueberry propagation: Researchers, students, berry growers. The users of the berry information range from home gardeners who are interested in growing bog blueberries in edible landscapes to small cottage industries based upon processed jam and jelly products.Controlled Environment: Owners, managers and employees of local horticulture operations and businesses; individuals considering potential horticulture production ventures; students at secondary and post-secondary levels including undergraduate and graduate students; initial and continuing training opportunities for the local workforce of horticulture operations. Chemistry: We are reasonably optimistic that the cumulative results of our work will be able to be submitted to the Journal of Natural Products. This is a peer-reviewed journal with a reasonably high impact factor (3.16) that is read by chemists and those interested in natural products having promising biological activities. This research on cancer and neuroprotection is very important to Alaska natives and all people living in the circumpolar north. The understanding of protective mechanisms of berries, a local food, is important for the berry and neutraceutucal industry. Another audience is the newly developing biotech industry in Alaska. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
These five projects addressed directives of our Governors since statehood to utilize natural resources to enhance the economy of Alaska and diversify it from extractive industries that dominate today. This work was a groundbreaking collaborative effort between the Departments of High Latitude Agriculture and Chemistry that provided an aggressive approach to bring ideas and products from basic research through demonstration to the consumer market. The idea was to develop a broadly based agribusiness sector in Alaska's economy based in the unique neutraceutical characteristics of Alaska's cultivated and wild harvested fruits and berries and products that can be manufactured from them. Success, directly dependent on funding has fallen short of those goals but the results to date are phenomenal. In the Propagation of four Alaska vaccinium species for potential field cultivation and wild stand enhancement best management practices were completed, demonstrated and shared with growers. Berry field research and development continues in controlled environments as well as field cultivation to develop best management practices for raspberries and currants for the fresh market. Chemistry determined that approximately 10% dry weight of Alaska blueberries to be nearly an order of magnitude higher that that found in commercial sources. Our research addressed questions regarding chronic illness such as metabolic syndrome, stroke and cancer that need both effective and economic therapies. We sought to understand at the molecular level 1) the interactions between blueberries and trophic factors related to cancer and stroke and 2) the effect that exposure to contaminants has on blueberry antioxidant effect. We continue to characterize blueberries in cell culture for chemoprotective effects. Suspicions were confirmed that it is in elevated levels in Alaska bog blueberries. Environmental toxins play a pivotal role in the initiation and progression of disease processes. Metals, pesticides, and halogenated biphenyls are suspected toxins both for the adult and developing nervous and immune systems. Chronic exposure, even at low levels, is inevitable due to increasing global industrial impacts. People living in a rural environment and/or those who rely on a subsistence life style (both prevalent in the State of Alaska) may be in great danger due to extremely slow transformation of contaminants in arctic regions and the resulting accumulation. Outreach: Chemistry: The research in this proposal has led to increased engagement of students in science courses. Adding lessons on blueberry chemistry has allowed incorporation of Alaska Native tradition ecological knowledge in our chemistry classes.Cooperative Extension: One of the challenges in making products from wild berries is the quality variability from year to year requiring recipe adaptation to produce a consistent product. Small producers developed products but often advertise them as "limited" in supply. The potential for these products is great demonstrating the need for best management practices for quality and quantity.
Publications
- Karlsson , M. and J. Werner. 2011. High tunnel covering materials for northern field production. Acta Horticulturae 893:1333-1339.
- Neely, H.L., R.T. Koenig, C.A. Miles, T.C. Koenig, and M.G. Karlsson. 2010. Diurnal fluctuation in tissue nitrate concentration of field-grown leafy greens at two latitudes. HortScience 45:1815-1818.
- Werner, J., Y. Okada and M. Karlsson. 2011. Using light emitting diodes in high latitude greenhouse production. Acta Horticulturae. 169.
- Duffy, L.K., Middlecamp, C.H., Godduhn A., Nicholas-Figueroa, L. and Fabbri, C.E. 2009. Because Science matters: engaging students in chemistry using traditional knowledge in Proceedings of "Lessons from continuity and change in the fourth international polar year", Fairbanks, AK.
- International Symposium on High Technology for Greenhouse Systems. 2009. Quebec City, Canada, P129, p. 156, GreenSys2009 Scientific Program (Abstract.)
- Holloway, P.S., K.M. Kokx, J. Auer and S. Pearce. 2009. Propagation of northern bog blueberry (Vaccinium uliginosum) by seeds and stem cuttings. Combined Proceedings International Plant Propagators' Society. 59:475-478.
- Holloway, P. S., K.M. Kokx, J. Auer and S. Pearce. 2009. Propagation of Alaska bog blueberry, Vaccinium uliginosum. HortScience. 44(4):1102. Abstr.
- Karlsson , M. and J. Werner. 2009. High tunnel covering materials for Northern field production.
- Karlsson, M. and J. Werner. 2008. Modified field environments for high latitude crop production, p. 64. 2008 International Meeting on Controlled Environment Agriculture, NCERA-101 North America Committee on Controlled Environment Technology and Use, Cocoa Beach, Florida.
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Progress 09/01/09 to 08/31/10
Outputs
OUTPUTS: Berry Propagation research developed a successful seed germination protocol, a stem cutting protocol for propagation of V. uliginosum and explored the variability in rooting by site and from plant to plant; and established permanent field plots of V. uliginosum and V. vitis-idaea at the Georgeson Botanical Garden for use in antioxidant, genetic, biochemical diversity research. Results were shared directly with individual berry growers, members of the Alaska Berry Grower Association and to the general public at a regional conference, Grown in Alaska, Palmer, Alaska; scientific poster was presented at the American Society for Horticultural Sciences national meeting, St Louis, MO, the International Plant Propagators' Society meeting, Denver CO and a public interest leaflet was revised to reflect recent research. In the controlled environment work four commercially available greenhouse films were tested using high tunnel structures. The evaluated materials were K50 Clear, K50 IR/AC, KoolLite380 and Solatrol. Raspberries and strawberries were grown to evaluate crop response. Long cane Tulameen were planted using 11.3-liter (3 gallon) large containers for immediate first season raspberry production. Excellent pollination and growth resulted in top quality fresh market raspberries in all environments including the field. More than 100 high quality marketable raspberries were harvested from each single cane plant. Disease inhibition due to blueberry interaction studies continue with an assay developed for translocation in myocytes and white blood cell cultures to screen the blueberry fractions. This allowed us to extend our knowledge of blueberries beyond inflammation and cancer to another chronic syndrome. Identification of bioactive nutrients in the diet that impact the metabolic state leading to improvement in chronic diseases like metabolic syndrome is important. Blueberry fractions may have selective effects on insulin sensitivity. Nutritional intervention has been shown to ameliorate some insulin resistance in type 2 diabetes. The results of chemical analysis of antioxidants research have been disseminated to a broad community through several mechanisms: PhD Thesis (May 2010) by Dr. Colin McGill entitled 'Biologically Relevant Secondary Metabolites of Vaccinium Uliginosum: Bioassay-Directed Natural Products Identification of Anti-Neuroinflammatory Agents in the Alaska Bog Blueberry'; two undergraduate research poster sessions; an honor's project report submitted to the Honors program Spring 2011; a poster presentation made Spring 2010 by Colin McGill at the UA Biomed Research Conference (UBRC) at UAA; and a seminar presented to the Biology Department at UAA. A request was made for INBRE support looking at the inflammatory mechanisms of iqmic and oral tobacco in oral epithelium and the protective effects of whole Alaska blueberry extracts and blueberric acid in general. PARTICIPANTS: Patricia Holloway, Professor of Horticulture, AFES, UAF; Alaska Berry Growers Association; Commercial berry producer, Brian Olsen, Kenai. Growers in Prince of Wales Island who have successfully propagated V. ovalifolium by air layering. Two growers in the Interior, Charles Knight and Papa Meneur, propagated thousands of seedlings of V. vitis-idaea and V. uliginosum and have transplanted them into cultivated fields. Two graduate students, Tina Buxbaum, and Rehanon Pampell, are working on pollination biology and bumble bee biology to enhance fruit set in wild and cultivated stands using native pollinators. We have developed a collaborative team with USDA ARS entomologist, Dr. Alberto Pantoja, UAF entomologist, Dr. Derek Sikes, and Alaska beekeeper, Stephen Peterson, to expand knowledge of native pollinators. Meriam Karlsson, Professor of Horticulture, project manager, Controlled Environment Laboratory, AFES, UAF; Jeff Werner, Research Professional; Jeffrey Johnson, Research Associate; Terry Marsh, Research Associate; Katherine Albers, Research Associate; Chris DuBois, Research Associate; Yosuke Okada, B.S. student; James Ward, B.S. student; Andrew Winkelman, B.S. student; Patrick Terra, B.S. student; Patrick Sanders, B.S. student; Justin Hogrefe, B.S. student; Melissa Gagnon, B.S. student; Kate Fournier, B.S. student; Shawn Biessel, B.S. student; Ryan Sloger, B.S. student; Chena Hot Springs Resort, Collaborator, Pike's Waterfront Lodge, Collaborator Dr. Duffy, Chemistry professor at UAF, led the project while Dr. Dunlap, a research assistant professor, managed and maintained the lab and cell cultures. A high school teacher received training on the ELISA technique which she will use with her students on high school research projects. A graduate student, Linda Nicholas-Figueroa, also participated in the design of the modifications for year 2 of this project. Dr. Tom Clausen, Chemistry Professor, in the natural products laboratory supported two students on this project. Colin McGill received his PhD in May 2010 for his contribution to the project (primarily the initial bioassay driven isolations and structural identifications). Jennifer Chambers is an undergraduate upper division chemistry major who developed an HPLC method for quantifying blueberric acid as well as our improved method to isolate it. TARGET AUDIENCES: Blueberry propagation: Researchers, students, berry growers. The users of the berry information range from home gardeners who are interested in growing bog blueberries in edible landscapes to small cottage industries based upon processed jam and jelly products. Controlled Environment: Owners, managers and employees of local horticulture operations and businesses; individuals considering potential horticulture production ventures; students at secondary and post-secondary levels including undergraduate and graduate students; initial and continuing training opportunities for the local workforce of horticulture operations. Chemistry: We are reasonably optimistic that the cumulative results of our work will be able to be submitted to the Journal of Natural Products. This is a peer-reviewed journal with a reasonably high impact factor (3.16) that is read by chemists and those interested in natural products having promising biological activities. Nearly all submissions report newly reported compounds with important biological implications. Blueberric acid falls under this heading in that is has not been reported before as a natural product, it has demonstrated ability to protect against oxidative stress (Kuhn's work), and it is a major component in a common berry consumed by much of the public (up to about 10% the dry weight of the berry). It is also an interesting secondary metabolite in that it is not derived from any of the major pathways (shikimates, polyketides, terpenes) but rather is an offshoot of primary metabolism. The incidence of brain cancer in Alaska is higher than the national average. This research on cancer and neuroprotection is very important to Alaska natives and all people living in the circumpolar north. The understanding of protective mechanisms of berries, a local food, is important for the berry and neutraceutucal industry. Another audience is the newly developing biotech industry in Alaska. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Berries grown under high latitude conditions have elevated concentrations of health promoting phytochemicals. Summer conditions with long twilight extended days, seasonal moderate temperatures and harsh winters appear to promote the synthesis and accumulation of antioxidants and other compounds of high nutritional value. Berries are harvested from wild stands but more intensely managed production systems are desirable to meet the dietary demands and marketing opportunities for quality berries. To decrease field uncertainty and improve seasonal productivity, high tunnels are now commonly used to produce a variety of crops. Traditional ethylvinylacetate (EVA) plastics such as K50 Clear are most commonly used in high tunnels and plastic greenhouses. Using alternative materials to accentuate or isolate patterns in the exceptional light and temperature conditions of high latitudes may improve berry growth and offer insights to factors significant for the nutritional quality. Understanding growth and development will be useful for improving field production and for conditioning propagation materials for introduction to field environments. In the disease inhibition due to blueberry interaction studies, we found that the SH-SY5Y neuroblastoma cells have measurable protein levels of the vascular endothelial growth factor and interleukin-1 receptor antagonist. Addition of BB fraction 8 to neuroblastoma cells inhibits the TNF-α related secretion of VEGF suggesting a potential reduction in cell division and blood vessel formation. BB fraction 8 induced the highest levels of IL-1Ra which suggest that IL-1Ra is involved in the regulation of VEGF secretion. In the chemical analysis of antioxidants research an HPLC method was developed for quantifying blueberric acid in berries using a specialized prevail organic acid column (Alltech) with a mobile phase of aq HCl (pH = 1). Our suspicions were confirmed that it is in elevated levels in Alaska bog blueberries. We determined it's concentration of about 10% the dry weight of our berries to be nearly an order of magnitude higher that that found in commercial sources. We also developed an improved method for isolating blueberric acid. Our earlier method relied heavily on silica gel chromatography using a chloroform / methanol solvent (9:1). Blueberric acid could be separated in this manner from fructose and malic acid only after repeated chromatographic separations. The old method was consequently expensive, generated large amounts of hazardous waste, and slow. The method we found that works the best is to run an extract rich in blueberric acid through and ion exchange column (weakly basic tertiary amine) using deionized water as an eluant until all the non ionic components (mainly fructose) have eluted. The acidic components (mainly blueberric and malic acids) are then flushed from the column as their ammonium salts with 5% ammonia solution. The free acids can then be obtained by running the eluant through a strongly acidic ion exchange column. Repeated chromatography is unnecessary as well as the need for using hazardous organic solvents.
Publications
- Duffy, L.K., Middlecamp, C.H., Godduhn, A. and Fabbri, C.E. 2009. Using Culture, Policy and Traditional Knowledge to Improve Engagement in Science Courses. Amer. Journ. Applied Sciences, 6(8): 1560-1566.
- Duffy, L., Middlecamp, C., Godduhn, A., Fabbri, C. and van Muelken, M. 2010. Chemistry, Society, and Civic Engagement (Part 3): Adapt and Adopt. Journal of Chemistry Education.
- McGill, Colin, 2010 Ph.D. Thesis: Biologically Relevant Secondary Metabolites of Vaccinium Uliginosum: Bioassay-Directed Natural Products Identification of Anti-Neuroinflammatory Agents in the Alaska Bog Blueberry.
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Progress 09/01/08 to 08/31/09
Outputs
OUTPUTS: Progress in propagation and production was made through the establishment of permanent field plots of V. uliginosum and V. vitis-idaea for use in antioxidant, genetic, biochemical diversity research. Graduate students work on the pollination biology of V. uliginosum and bumble bee biology. A retractable double-layer flat-roof high tunnel has been erected to protect the crop from inclement weather while taking full advantage of solar heating, natural ventilation, and the optimal light and temperature conditions of the intense northern field season. The Alaska Berry Growers (ABG) and public have been kept informed through emails and educational meetings. The Chemistry Ph.D.graduate student has completed the research and thesis and defense are expected Spring 2010. A publication will be submitted to the Journal of Natural Products. Ongoing collaboration with Dr. James Joseph (USDA Human Nutrition Research Center at Tufts University) accomplished a significant technology transfer. Sally Gustafson (PhD student, UAF) established microscopy-driven investigations on the effects of blueberry extracts on lipid microdomain (raft) formation in neuronal cells during a 4-month residency in Dr. Joseph's laboratory and returned with technological know how regarding microglia (BV-2 cells), primary hippocampal neuron cultures, and the fractionation of blueberry extracts according to molecular families. Additional connections through this research were established with Dr. Donald Ingram, a researcher at Pennington Biomedical Research Center (formerly NIA), and Dr. Wilhelmina Kalt (Agriculture Canada). Research was disseminated through invited presentations at national and international meetings at Tufts University, and to the Economic Development Task Force of Fairbanks, AK. Significant advances were accomplished through participation of Dr. Kuhn in the University Economic Development Work Group to alert the community of the commercial opportunities with respect to Alaska Blueberries. Through support and guidance by the UAF technology transfer officer, the PI was able to research the interest potential of national businesses in AK blueberries or derived nutraceuticals at the International Food Technology Conference (Anaheim, CA, 2009), and steps are initiated to secure intellectual property. These efforts have led to the development of an 2010 SBIR application. Samples to determine the antioxidant efficacy of blueberry extract or polyphenols using TAP assays with a focus on regional variation and level of contaminants have been collected. The TNF stimulating assay has been perfected and chemoprotective fractions identified to determine the interaction of blueberry extracts and polyphenols in the presence of mercury and PCB by measuring CRP and IL-6 in a cell culture system. VEGF levels have been measured in our cell culture system. We have verified preliminary results indicating chemoprotection to SH-SY5Y neuroblastoma cells (a cancerous cell line) exposed to an inflammatory stimulus. After cells were exposed to stimulus, some blueberry extract fractions inhibited the release of IL1, ILRA and VEGF. PARTICIPANTS: Carol E. Lewis, Director AFES, Project Oversight. PROPAGATION: Patricia Holloway, Professor of Horticulture, Research Technician Grant Matheke. Two graduate students, Tina Buxbaum, and Rehanon Pampell, working on pollination biology and bumble bee biology to enhance fruit set in wild and cultivated stands using native pollinators. COLLABORATORS: Alaska Berry Growers Association, Kenai, commercial berry producer, Brian Olsen, Kenai, growers in Prince of Wales Island, two growers in the Interior, Charles Knight and Papa Meneur, and collaborative team USDA ARS: entomologist, Dr. Alberto Pantoja, UAF entomologist, Dr. Derek Sikes, and Alaska beekeeper, Stephen Peterson to expand knowledge of native pollinators. CONTROLLED ENVIRON: Meriam Karlsson, Professor of Horticulture, project manager; Jeff Werner, Research Professional; Jeffrey Johnson, Research Associate; Graduate Student: Yosuke Okada. Undergraduate students: James Ward, Andrew Winkelman, Patrick Sanders. COLLABORATORS:Chena Hot Springs Resort, Collaborator, Pike's Waterfront Lodge, Collaborator, Rough Brothers Greenhouse Manufacturing Company (Cincinnati, Ohio), Collaborator, Link4 Corporation (Placentia, California), Collaborator, Svensson Screens (Charlotte, North Carolina), CHEMICAL ANALYSIS: Thomas Kuhn Associate Professor, in close collaboration with Thomas Clausen Department Chair and Professor of Chemistry and Biochemistry and Lawrence Duffy, Professor of Chemistry and Biochemistry. Fractionation of AK wild bog blueberries was accomplished and driven by his graduate student Colin McGill, who graduated recently with a Ph.D degree in biochemistry based on his investigations .Sally Gustafson, PhD graduate student, succeeded in demonstrating the capacity of blueberries and purified compounds to potently inhibit NOX. Dr. Kriya Dunlap has accomplished the fractionation of an nSMase inhibitor from AK wild bog blueberries in collaboration with Dr. Colin McGill. Her efforts were recognized by the Dean of the College of Natural Sciences and Mathematics. She was promoted to a Research Faculty at UAF providing her status as an independent principal investigator. Sigourney Walker (Undergraduate student) participated in many of the isolations including IV, fructose, and cyaniding. Talal Ahmed, Gavin Baker, Patrick Bollig, Jennifer Chambers, Carrie Duplisea, Christopher Ebner, Kathleen Gerdes, Andy Lester and Brad Weis (Organic Laboratory {Chem 324W} students) performed the first two steps in the synthesis described. COLLABORATORS: Dr. James Joseph, USDA Human Nutrition Research Center at Tufts University, Dr. Donald Ingram, Pennington Biomedical Research Center (formerly NIA), and Dr. Wilhelmina Kalt,Agriculture Canada. A new collaboration was established with Dr James Bamburg, Colorado State University. The goal of the interaction between the two laboratories is to investigate whether AK wild bog blueberries, fractions derived, and purified compounds harbor the potency to rescue neurons from a reorganization of their actin cytoskeleton into rod-like structures. Bamburg and co-workers demonstrated the formation of rods as an early event in neurodegeneration. TARGET AUDIENCES: PROPAGATION: Home gardeners, small cottage, Alaska Berry Growers Association, researchers. CONTROLLED ENVIRONMENTS: Owners, managers and employees of state-wide commercial greenhouse and other horticulture operations and businesses; berry and specialty crops growers, individuals considering or participating in horticulture production ventures; students in secondary and post-secondary educational programs and college level undergraduate and graduate students; initial and continuing training opportunities for the local workforce of horticulture operations, plant physiologists, climate change and climatology researchers. CHEMICAL ANALYSIS: Knowledge obtained through this research will benefit three principal audiences. First, inclusion of nutritional aspects with respect to biomedical research continues to become an important research direction in the molecular biosciences at UAF and a key research subject in Biochemistry. Second, the potential of this research to foster nutraceutical business endeavors is currently discussed with the university and the business community of Fairbanks. This research demonstrates a new dimension to natural resource use in Alaska. Third, this research continues to demonstrate that nutrition could provide a powerful approach to alleviate disease and aging-derived deficits to CNS health, which would benefit the public at large. Key to this research is the demonstration that natural compounds not only act as diffuse scavengers yet rather target distinct and defined biochemical entities in complex cellular signaling network. Therefore the benefits of nutrition are demonstrated to reach far beyond antioxidant capacity. The incidence of brain cancer in Alaska is higher than the national average. This research on cancer and neuroprotection is very important to Alaska natives and all people living in the circumpolar north. The understanding of protective mechanisms of berries, a local food, is important for the berry and neutraceutucal industry. Another audience is the newly developing biotech industry in Alaska. PROJECT MODIFICATIONS: PROPAGATION:The focus of this research has been narrowed to include two species of Vaccinium, V. vitis-idaea and V. uliginosum, in part because of requests from constituents and also because of climactic challenges working in Interior, Alaska. Some of the coastal species of Vaccinium cannot survive in Interior, Alaska, and a suitable location has not been found for field trials in coastal Alaska. The two remaining species have the greatest economic potential in Alaska. Whereas, the species have been narrowed, the scope of the project has expanded to include important components to management of wild and cultivated stands including methods of field establishment and pollination biology for improved fruit production. Both of these areas were pursued at the request of the Alaska Berry Growers Association. CHEMICAL ANALYSIS: Modification for Year 2 Plan: Besides measuring IL-6 and CRP in our assay system, we will also assay for Il-1 beta so we can complete the total IL-1 screening. We will also develop an assay for Glut 4 translocation in myocytes and white blood cell cultures to screen the blueberry fractions. This will allow us to extend our knowledge of blueberries beyond inflammation and cancer to another chronic syndrome. Identification of bioactive nutrients in the diet that impact the metabolic state leading to improvement in chronic diseases like metabolic syndrome is important. Blueberry fractions may have selective effects on insulin sensitivity. Since our year 1 points to some blueberry fractions interfering with the release cytokines, we propose to develop a translocation assay using Glut 4 levels in myocytes and white blood cells.
Impacts
Small markets continue to develop around a cottage industry of wild jams and jellies. Wild blueberries are sold at all 19 of Alaska's farmer's markets. All major jam and jelly businesses in Alaska include at least one wild blueberry product in their lines. One market emphasizing the phytonutrient value of wild Alaska blueberries is continuing development in Kenai, Alaska. Demand for freshly harvested wild berries continues to grow, although the exact size of the industry is not known. Demand for basic information on berry culture is high, we are working with the Cooperative Extension Service and the Alaska Division of Forestry to provide information via a statewide conference and publications in 2009. Summer conditions of twilight extended days, seasonal moderate temperatures and harsh winters appear to stimulate the synthesis and accumulation of antioxidants. Understanding the environmental conditions and the plant physiology governing the formation of health promoting substances is necessary to produce berries of highest possible nutritional quality. These research findings and observations support the continuous development and advancement of producer guidelines and the best management procedures for field establishment, production management and harvest. The long-term goal of our chemistry research centers on the containment and reduction of CNS inflammation through natural products to alleviate neurodegeneration and its dire consequences. Specifically we proposed to elucidate the molecular mechanisms conferring neuroprotection against inflammatory processes and oxidative stress by molecular compounds in Alaska Wild Blueberries and to characterize these natural compounds as a lead drug development approach to enhance therapeutic or preventative strategies. Our findings that AK wild bog blueberry extracts contain highly specific inhibitory potency against the ceramide-producing nSMAse and the ROS-producing NOX, were received with great interest by the scientific community. We demonstrated not only the non-antioxidant nature of these inhibitory compounds in blueberry extracts but, more importantly, their discrete partition in fractionation procedures. As one of few laboratories, we were able to retain the potency of crude whole blueberry extracts subjected to fractionation. This implies that berry fruit are likely to contain inhibitory compounds of high selectivity and potency against molecular targets in neuronal cells. It is noteworthy that these inhibitory compounds and fractions exhibit higher potency, increased specificity, and virtually no toxicity and thus outperform current pharmacology. Lipid rafts harbor a vital role in the generation of intracellular signals that emerge at the plasma membrane and in particular redox signals. Our research demonstrates that blueberry extracts, specific fractions, and purified compounds have the capacity to modulate raft dynamics and thus disable NOX assembly. These studies demonstrate that natural compounds in blueberries physically alter the plasma membrane environment and thereby support the survival of neuronal cells.
Publications
- Duffy, L.K., Dunlap, K.L., and Nicholas-Figueroa . 2009. Chemo-protection for brain cancer by Alaska blueberries. 2009 Arctic Science Conference abstract.
- Holloway, P. 2009. How to germinate seeds of Alaska wild blueberries and lingonberries (lowbush cranberries). Georgeson Botanical Notes No 34. (revised) 2p.
- Holloway, P. S., K.M. Kokx, J. Auer and S. Pearce. 2009. Propagation of Alaska bog blueberry, Vaccinium uliginosum. HortScience. 44(4):1102. Abstr.
- Karlsson, M. and J. Werner. 2010. High tunnel covering materials for northern field production. Acta Horticulturae
- Kuhn, T. 2009. Invited presentation: Winter Conference on Brain Research 2009. Kuhn, T. 2009. Invited presentation: Berry Health Monterey 2009, International Society for Nutraceutical and Functional Foods, 2009.
- Kuhn, T. 2009. Invited presentation: Tufts University.
- Kuhn, T. 2009. Invited presentation: Economic Development Task Force of Fairbanks, AK.
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