Berry Flavonoids - Structure, Bioactivity, and Interactions

Berry fruits have shown a remarkably diverse range of bioactivities relevant to human health protection. The health-promoting potential of berries include antioxidant properties, anticancer, anti-adhesion (prevention of urinary tract infection, antiinflammatory, and neuroprotection properties. These broad spectrum health-protective properties are attributed to the distinctive and multifaceted mixture of phytochemical components that naturally accumulate in berry fruits. Natural potentiation between interacting components which co-occur within the fruits can intensify the potency and broaden the scope of protection beyond that of any single component. Our lab has concentrated on elucidating the chemopreventive and antioxidant capacity of berry fruits, including efficacy in multiple stages of the carcinogenesis process. In addition, we have examined the adaptogenic potential of fruit components; their capacity to increase endurance and stamina in test subjects.

In order to characterize the components responsible for health benefits in berry fruits, the components must be carefully separated to allow clean bioassay and characterization. Vacuum chromatography is performed first on a Toyopearl gel matrix, which provides for simple and efficient separation of sugars from other compounds in the extract. Other substrates such as silica gel are commonly used for subsequent rounds of subfractionation. One of the important pursuits in our team is to determine the effect(s) of alternative extraction and fractionation tactics on the integrity and structure of the components from fruits/functional foods, as many natural plant compounds may be ephemeral in nature, and easily degraded during the process of phytochemical separation and analysis. In addition, many plant-produced bioactive compounds are difficult to elucidate due to their large, complex, polymeric structures and potentiating interactions between phytochemicals that co-occur in the plant host.

Fractionations are typically monitored using TLC on silica gel plates. This protocol permits the separation of intractable and complex proanthocyanidin oligomers and other complex bioactive compounds with minimal loss of structural integrity, and determination of their bioactivity. We accomplish structural characterization of active constituents using an array of phytochemical detection strategies (HPLC, TLC, Mass Spectroscopy and NMR), and simultaneously, via bioassay-guided fractionation, elucidate the interactions between potentiating phytochemicals that co-occur in a bioactive edible plant.
In vitro cell culture systems

Plants continue to provide novel leads for multifaceted drugs to combat complex human chronic diseases, however, full appreciation for the diversity of compounds available through natural plant sources has not been realized. Development of a streamlined, scaled-up, reproducible plant cell culture model system for uniform production and harvest of bioactive components from plant sources provides a means to decipher the mechanisms of action specific to complex natural phytochemicals, to determine the degree of interaction between components in a food matrix in terms of disease intervention, and to track the metabolic route of bioactive components after ingestion by animal or man. The chemical composition of plants collected from the wild varies significantly and unpredictably, and often these plant collections lead to habitat disruption and political controversy. Uniform, continuous plant cell cultures maintained under controlled conditions do not have these disadvantages.

Additional advantages inherent to the cell culture models as research and discovery tools include:

* Phytochemical production can be more reliable, simpler, and predictable
* Phytochemical production is highly inducible by elicitation
* Isolation of the phytochemical can be rapid and efficient, as compared to extraction from complex whole plants
* Compounds produced in vitro often directly parallel compounds in the whole plant
* Interfering compounds that occur in the field-grown plant can be avoided in cell cultures
* Cell cultures can yield a source of defined standard phytochemicals in large volumes
* Cell cultures can be radio-labeled such that the accumulated secondary products, when provided as feed to laboratory animals, can be traced metabolically

Reliable, high volume, uniform cell suspension cultures from the genus Vaccinium have been adapted to continuous cell culture systems. The cell culture model, on a 2 week rotation, can be induced to produce the same range of bioactive plant phytochemicals as would be produced by their parent (donor) plant, and provides a streamlined vehicle for extraction of bioactives, a controlled system for elicititation, and a means to investigate the potentiating effects of interacting substances in a bioactive mixture. Tomato cell culture systems for elicitation of lycopene and its precursors, and soy, kudzu, and red clover systems for elicitation of isoflavone metabolites have similarly been developed in our laboratories. Part of our strategy is to determine elicitation protocols (chemical and/or physical stimuli) which result in maximal yields and most desirable chemical profile for secondary medicinal products from cell cultures

Nutrasorb - Invented by Nature, Improved by Science

A novel project within the Lilalab is evaluating the functional and technical parameters surrounding Nutrasorb™, an innovative and cost-effective platform to manufacture good-tasting, nutritious, and shelf-stable functional foods and ingredients. Nutrasorb™ is designed to deliver doses of phytonutrients within food structures to maintain health and wellness.

The matrix is an all-GRAS (Generally Recognized as Safe) food material, effectively absorbing and concentrating phytonutrients and bioactive natural products from plant juices, teas, and extracts. The enhanced matrix is then able to be processed into a wide variety of food or beverage products with enhanced nutritional value. The Nutrasorb™ matrix enables higher amounts of phytonutrient delivery compared to conventional plant servings.

For more information, visit http://nutrasorb.com.

 

The Global Institute for BioExploration (GIBEX)

The Global Institute for BioExploration (GIBEX) is a global research and development network that promotes ethical, natural product-based pharmacological bioexploration to benefit human health and the environment in developing countries. GIBEX was established by Rutgers, the State University of New Jersey, the University of Illinois at Urbana-Champaign, and recently North Carolina State University. All are leading US universities with strong records of building successful international programs in discovering and developing life-saving medicines.

GIBEX is guided by the pioneering “Reversing the Flow” paradigm intended to bring pharmacological screens to developing countries (Screens-to-Nature technology) and reverse the human ‘brain drain’. GIBEX does not remove any natural resources from partnering countries. Instead, it trains and equips local scientists and students with innovative, cost-effective, and portable drug-discovery tools and technologies that can be directly deployed into forests, savannas, deserts and marshes. In addition to adapting the process of drug discovery and commercialization to the needs of partnering countries, GIBEX promotes sustainable infrastructure and capacity improvements, local intellectual property ownership, conservation, educational opportunities, and entrepreneurship.

'REVERSING THE FLOW' concept is a founding principle of GIBEX, which distinguishes it from other international bio-exploration activities.

• Reversing the Resource Drain
  Biological materials are customarily collected from the natural sources in developing countries and moved to developed countries for pharmacological screening. This practice creates numerous political, social and legal problems. GIBEX reverses the flow of the discovery process. It brings portable, field-deployable pharmacological screens directly to nature in the developing countries, thus creating a new paradigm of ethical bio-exploration.
• Reversing the R&D Drain
  Developing countries are rarely involved in pre-clinical and clinical development of therapeutic agents derived from their flora and fauna. GIBEX enhances the economic returns for the source countries by keeping the later-stage development work in these countries through building, nurturing and facilitating internal R&D. Thus, the value of any licensing agreement for the source country is maximized. To achieve this goal, GIBEX is building pre-clinical and clinical development capacity in the member countries using the 'reversed brain drain' approach (see below).
• Reversing the Brain Drain
  The training of the scientists from developing countries is customarily done in the developed countries. This practice, in addition to being costly and inefficient, creates a major 'brain drain' detrimental to the local economies. It makes poor countries poorer and the rich ones richer. GIBEX sends scientists from developed countries to developing countries to provide hands-on training in the application of the 'screens to nature' technology as well as in all other aspects of pharmacological discovery and research.

For more information, visit the GIBEX website.

Investigating Tropical Plants for Novel Antimalarial Compounds

Working with the Gates Foundation's Medicines for Malaria Venture, the Lilalab has been working in conjunction with Rutgers University as well as the University of Cape Town in order to uncover new candidates in the fight against malaria.

Looking at the South American vine Canella winterana, Dr. Grace has uncovered a set of sesquiterpenes that provide activity against the malaria parasite in a cell culture model. Her work also set up the structural characterization of these compounds, which are undergoing further testing. This is the first example from the MMV project of new antimalarial compounds from plant sources.

Catalyst for Science Education and Validation of Traditional Knowledge

In this new multi-year project from the USDA Higher Education Challenge (HEC), the Lilalab is collaborating with education experts at North Carolina State University, Rutgers University, the University of Alaska Anchorage, the University of Alaska Fairbanks, United Tribes Technical College and Native American students to develop a lab and lecture curriculum based upon the Screens-to-Nature technology that GIBEX has pioneered. From the grant summary:

Native Americans students, dramatically underrepresented in science, technology, and advanced education, are introduced to straightforward, logical and illustrative field deployable portable biodiscovery assays known as Screens to Nature (STN) technology, and thoroughly coached on the use of these renewable and safe-to-use toolkits to investigate the biological activity of indigenous plants, fungi, microbes and marine organisms in their own communities.  The inventive STN portfolio is designed to help students recognize the unique bioactive (human health-protective) properties of their local natural resources, to put them in direct charge of making novel discoveries using the scientific method, to capture and sustain interest and enthusiasm for science discovery and to concurrently validate the traditional ecological knowledge of the community elders through science principles. 

STN is simultaneously a learning tool, and a community mobilization technique that promotes interaction among different stakeholder groups (tribal leaders, elders, youth, etc.) and uniquely connects the communities, environment, and health.  The experiences will develop future scientists of Native American origin as local ambassadors, with an invigorated sense of community citizenship.  Their instructors will also engage in a transdisciplinary summer training institute designed to amplify their appreciation for integrated scientific instruction and experience first hand the next steps after initial field bioassay discoveries, through validation of bioefficacy in the laboratory and clinic..  Our long terms goals are to create a self-sustaining, timely, and cost-effective resource to link young adults and elders in tribal nations, expand and enliven educational concepts, build pride and community, scientifically validate traditional ecological knowledge, and reinforce appreciation and recognition across generations.