Recipient Organization
ALABAMA A&M UNIVERSITY
4900 MERIDIAN STREET
NORMAL,AL 35762
Performing Department
Food & Animal Sciences
Non Technical Summary
Diabetes is a major epidemic in the US and worldwide. The rise in diabetes is attributed to the rise in obesity as diabetes is a major health complication. Natural and alternative approaches are needed for the prevention and cure of diabetes. This project is focused on evaluation of the potential health benefits of selected spices and plant food components in the prevention of obesity induced diabetes. A multidisciplinary approach will be used to determine the anti-diabetic potential of spices and to disseminate the knowledge. Dietary modification and diet management is a major concern among diabetic individuals and this project will evaluate consumer's knowledge about health properties of spices and foods, as well as provide participants with nutrition education needed for the effective management of diabetes. Adolescents have poor dietary habits and generally consume calorie dense foods which may lead to obesity. Health enhancing functional foods are needed to increase healthy food choice. The development of functional foods as a part of this project with spices would lead to wider selection of spice containing foods not only to enhance flavor but to increase anti diabetic potential. Collaborative efforts between food scientists and nutritionists are needed to ensure successful formulation of functional foods that the consumer deems safe, appetizing and beneficial to maintaining good health and the collaboration of nutritionists to transfer the information to the consumers via nutrition education modules.
Animal Health Component
75%
Research Effort Categories
Basic
15%
Applied
75%
Developmental
10%
Goals / Objectives
Objective 1: To screen for foods (selected spices and plant extracts) with components that possesses antioxidant and antidiabetic potentialObjective 2: To evaluate the effect of selected foods (selected spices) in reducing blood glucose level in streptozotocin treated rats along with high sugar and fat diet induced pre and post diabetic model.Objective 3: To conduct a nutrition education intervention program to impart healthy life style and changes dietary behavior of pre and diabetic adolescents Objective 4: To develop functional food products by incorporating selected foods (spices and plant extracts) with anti-diabetic potential to promote healthy eating
Project Methods
Materials and methods: (Objective 1); In vitro studies and cell culturePowdered spices (turmeric, ginger, garlic), selected fruit (Guava) and aloe-vera will be purchased from Monterey Bay Spice Company, (Watsonville, CA) and local grocery stores. Spices will be treated thermally (30°C, 40°C, 50°C, 60°C) to optimize extraction of bioactive components.Antioxidant potential of extracts: Extracts will be tested for total phenolic content by Folin-Ciocalteau (FC) methodandotal flavonoid content in extracts will be determined. Identification and quantification of various chemical compounds present within extracts will be analyzed by HPLC (Revathy et al 2011 & Bocchini et al 2001). The antioxidant activity of extracts will be characterized in variousin vitromodel systems such as DPPH radical scavenging, Ferric reducing antioxidant power oxygen radical absorbance capacity (ORAC), trolox equivalence antioxidant capacity assay (TEAC) (Miller et al., 1993), total antioxidant capacity (TAC) , and nitric oxide radical scavenging activity (NORS) , oxidation of liposomes. Inhibition potential of extracts against carbohydrate and lipid metabolizing enzymes will be determined by following in-vitro protocols. α-glucosidase, α-Amylase inhibition and Pancreatic lipase inhibition by extracts will be determined .Cell Culture HIT-T15, Hep G2, 3T3L1 cell lines will be obtained from the American Type Culture Collection (Manassas, VA). They will be grown in appropriate media and maintained in an incubator at 37°C and humidified 5% CO2 atmosphere. Cells will be sub-cultured once a week and will be treated with different concentrations of extracts at (100, 200, 400, 800, 1000 µg/ml) at 37°C for 24 hours. Cells will then incubated with 25 µM glucose and incubated for another 24 hours at 37ºC. Hydrogen peroxide (400 µM) will be added three hours prior to the harvesting time. Cells will be lysed by using ice-cold solubilizing buffer and centrifuged at 10,000xg for 20 min at 4ºC. Collected supernatants will be analyzed for antioxidant activities. The effect of extracts and glucose on cell viability will be determined according to Abnova MTT (3-(4, 5-dimethylthiazolyl-2)) cell viability assay kit (Walnut, CA). A LDH cytotoxicity detection kit (Clonetech Laboratories, Inc. Mountainview CA) will be used according to the manufacturer's instructions to determine cytotoxicity of extracts. Microscopic examination of cell morphology will be performed using Giemsa stain and fixed with methanol. Formation of malondialdehyde (Cayman's TBARS (tiobarbituric acid) assay kit (Ann Arbor, MI)) and formation of Reactive oxygen species (ROS) will be determined using the oxiselect intracellualr ROS assay kit. Induction of antioxidative enzymes, Superoxide dismutase (SOD), Catalase (CAT) activity, Glutathione peroxidase (GPx) activity will be determined using a Cayman Chemical assay kit (Ann Arbor, MI). Antioxidant, Reduced glutathione (GSH), levels will also be measured using a Cayman Chemicals assay. Anti-inflammatory enzyme, Cyclooxygenase (COX-2), activity will be determined by using Enzo Life Sciences assay kit (Farmingdale, NY).Objective.2:Animal model: Experiment design:In vivo diabetic & pre-diabetic models will be utilized for the assessment of antidiabetic potential of food components at selected concentrations with feeding high fat (40% of calories)and sugar diets (40% of calories). Eight week old male wistar rats (48) will be purchased form Harlan laboratories (Harlan (Envigo), IN, USA).Sample collection: Blood will be drawn to document serum glucose levels. Animals will be cut open for collecting samples. Blood will be drawn by cardiac puncture. Samples (liver, pancreas) will be collected and saved for further analysis.Prior to euthanasia an oral glucose tolerance test (OGTT) will be performed at the end of the treatment. After 12 hours of fasting, glucose (2 g/kg) will be administered by gavage. Blood samples will be taken from rodent tail at 0, 30, 60, and 120 min after administration. Blood glucose levels will be measured using glucose meter.Biochemical analysis of serum and liver:During autopsy, whole blood taken from rats from each group was allowed to clot and the serum separated and analyzed for alanine aminotransferase (ALT), and Gamma-glutamyl Transferase (GGT) using standard kits (Cayman Chemicals Ann Arbor, MI, USA; Biovision). Triglycerides, Glucose, total cholesterol, high density lipoprotein -cholesterol (HDL-C), Low density lipoprotein -cholesterol (LDL-C), Creatinine, will also be determined using standard kits (Cayman Chemicals Ann Arbor, MI, USA).Bone Mineral analysis:Femurs will be excised from rats and weights of bones, bone length, and diameter will be recorded. Selected minerals (Ca, P, Mg, Zn and Fe) will be analyzed by inductively coupled plasma (ICP) (AOAC 984.27).Detoxification Antioxidant Rat Enzyme Activity -Detoxification and antioxidant enzymes (GSH, GST, GPx, SOD, and Catalase) will be measured in this study. Liver samples from the rats will be used for evaluation. GSH, GST, GPx, SOD, and Catalase kits (Cayman Chemical Company, Ann Arbor, MI) will be used according to the manufacturer's instructions for determination of enzyme activity.Objective 3Study design and data collection -Data will be collected from students, faculty and staff for a randomized intervention trial to test a nutrition intervention study to improve nutrient and health knowledge as well to reduce risk of diabetes at Alabama A&M University. At baseline, over 400 individuals with or without diabetes will be enrolled in the study. Trained research personnel will collect data following standardized protocols at baseline and after the intervention period. Study protocols will be approved by the Institutional Review Board at the Alabama A&M University.Subjects -Individuals with signed volunteer forms for the study, complete data on socio-demographic characteristics, dietary intake and measured blood sugar, height and weight will be selected for nutrition intervention program (approximately 50). All protocols will be approved by the Institutional review Board (Human Subjects Committee). The participants will receive a stipend of $100 per year.Assessment of dietary intake -Food intake of participants will be assessed by the NIH Food Frequency self-administered semi-quantitative questionnaire.Dietary Intervention Study -A four-week educational community dietary intervention study will be conducted at Alabama A&M University. This intervention study will educate both diabetic and non-diabetic individuals on healthy eating, nutrient requirements, and component function and use of medicinal plants in disease prevention.Nutrition education intervention will include information related to the development of effective, minimal intervention, sustainable dietary strategies to maintain weight and achieve weight and minimize the risk of dangerous, and misleading dietary weight-maintenance/weight-loss practices. The educational component will be designed to be integrative. The major purpose will be to show how food choices and physical activity are tied to personal behavior, physical and emotional health, and the environment.Objective 4:Functional food products will be developed utilizing fruits, spices and other phytochemical rich foods to maximize their nutrition and health benefits to customers. The product will be designed to appeal as a self-serve snack with potential health benefits as a value added product. A market research survey will be conducted in order to assess the target market, suggested set price, flavor profile, and product texture. The product development will encompass ideation, concept development and testing, formulation, nutritional analysis, shelf life testing, development of a HACCP plan, packaging, and sensory testing.