Progress 10/01/17 to 09/30/20
Outputs
Target Audience:The project is to identify dietary compounds from byproducts of fruits and vegetables that have anti-obesity effect by activating a function of brown adipose tissue and investigate their molecular mechanism.Therefore, target audiences are lab students, scientists who works in nutrient metabolism related to obesity, obese people, and communities who works in food and nutrition such as school teacher and nutritionist in counties. Results and information that are generated through this project were delivered to lab students by laboratory instruction during performing experiments that are proposed in this project.The data was delivered by poster presentations to other scientists and students who works in nutrition and obesity research areas at a scientific conference (Annual American Society for Nutrition Conferences) and to school teacher and nutritionists in Annual North Carolina Association of Family & Consumer Sciences Conferences where North Carolina school teachers and nutritionists attend. The data was also delivered to other scientists as scientific journals. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?During the three years, three master students, one undergraduate student, one PhD student, and two postdoctoral research fellows received the training from the project by obtaining new knowledge and experimental skills. Courtney Richard, a URSP student received one-on one training with a mentor and other team members about how to extract bioactive compounds from mango seed and how to measure total phenolic content and antioxidant properties and obtained knowledge about antioxidants. Courtney Richard had one published journal article as a co-author and make one presentation at a scientific conference. Olaniyan Jamal Cash, a master student received one-on one training with a mentor and other team members about how to culture RAW macrophage cells and extract bioactive compounds from red-onion peel, and how to measure gene expression from the cells and obtained knowledge about adipocytes related to inflammation in obesity. He made one research presentation at a scientific conference. Salma Altuwaijri, a master student received one-on one training with a mentor and other team members about how to extract bioactive compounds from garlic scape and how to measure total phenolic content and antioxidant properties and obtained knowledge about antioxidant. Salma Altuwaijri earned a Master's degree and made one presentation at a scientific conference. Olugbenga Balogun, a master student received one-on one training with a mentor and other team members about basic experimental skills e.g. gene expression, cell culture and animal handling to perform experiments to examine functions of garlic scape and obtained knowledge about molecular nutrition and functional foods. Olugbenga Balogun earned a master degree with a thesis and then continually started a PhD program. Dammah Otieno, a PhD student received one-on one training from a mentor about experiments how to isolate primary adipocytes from brown and white adipose tissue of mice, respectively and culture them. She expanded her knowledge to adipocyte development such as Wnt signaling. Dammah Otieno earned her doctoral degree with a dissertation. She made five presentations at scientific conferences and obtained two first authored and one co-authored published journals. Dr. Kyoungsoo Ha, a postdoctoral research fellow worked in the lab for a short period. He was trained to extract samples and to perform western blot analysis to measure protein expression. Dr. Ha had one published journal as a co-author. Dr. Ya Pei, a postdoctoral research fellow had received one-on one training from a mentor and knowledge related to BAT functions and obesity-induced inflammation, which attain greater proficiency to be an independent scientist in molecular nutrition area. Dr. Pei has prepared three conference abstracts, presented at scientific conference, and one first authored published journal. How have the results been disseminated to communities of interest?The dissemination to communities of interest was through a Small Farm Field Day at North Carolina A&T State University. In June 14, 2018, we educated around 100 farmers on health effects and re-usable applications of red and yellow-onion peel and their tea by presenting a poster that was developed for the education purpose regarding "onion peel: reducing obesity and inflammation". Another dissemination to communities of interest was through NCAFCS annual conferences (2018 and 2019) by presenting posters about our scientific findings to conference attendances who are usually school teachers and local health related workers and educators. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Objective 1 has 8 tasks by collecting and preparing extracts of by-products, identifying dietary compounds, testing cytotoxicity of samples, culturing primary brown adipocytes and 3T3-L1 cells, treating samples into both cells, examining gene and protein expression in these cells, manipulating these cells using TNFα to mimic inflammatory conditions, and then measuring synergistic effects of dietary compounds to increase UCP1 expression. We made extracts of food by-products, garlic scape, mango seeds, watermelon rind, and brewers' spent grain (BSG) and cytotoxicity of these extracts were measured in 3T3-L1 adipocytes and primary brown adipocytes. Mango seed, watermelon rind and BSG extracts did not show any significant difference in both cells. 3T3-L1 cells that were treated with garlic scape extract showed reduced adipogenesis and lipogenesis and increased lipolysis through AMPK signaling pathway during the differentiation, but it does not develop beige adipocytes. Bioactive compounds in garlic scape will be identificated this year, this work has been delayed due to the COVID 19 situation. We found a mechanism that quercetin, a major bioactive compound of onion peel regulated non-shivering thermogenesis in brown adipocytes by beta-adrenergic receptor and MAPK signaling pathways. Quercetin reduced inflammation in brown adipocytes by regulating SIRT1 signaling pathway in TNFα-induced inflamed primary brown adipocytes. Objective 2 includes 8 tasks by feeding animals, collecting tissues and blood, analyzing lipids in blood and liver, measuring gene and protein expression, measuring body composition and energy expenditure, and analyzing histology, testing glucose and insulin tolerance, and measuring cecal microbiome. Mice were fed with low-fat diet (LFD) and high-fat diet (HFD) with or without 10% OR 30% BSG for 16 weeks and then glucose and insulin tolerance tests were performed. Glucose and insulin tolerance were not clearly changed in mice supplemented with BSG, but cholesterol concentrations in liver and plasma were reduced. The expression of genes related to cholesterol metabolism were changed in liver. Mice were fed with LFD and HFD with or without 1% quercetin for 16 weeks. Mice fed a HFD supplemented with 1% quercetin (HFDQ) had reduced body weight and total plasma cholesterol level. HFDQ-fed mice had decreased retroperitoneal white adipose tissue (RWAT) weight and browning effect and lipolysis were increased in RWAT. The expression of genes that are involved in the regulation of non-shivering thermogenesis in BAT were increased, but energy expenditure and core body temperature were not changed. However, HFDQ-red mice showed increased physical activity during dark periods at room and cold temperatures. Quercetin decreased the Firmicutes to Bacteroidetes ratio and increased short-chain fatty acid production in the feces of HFD-fed mice.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Pei, Y., Otieno, D., Gu, I., Lee, S., Parks, J. S., Schimmel, K., & Kang, H. (2021) Effect of quercetin on non-shivering thermogenesis of brown adipose tissue in high-fat diet-induced obese mice, The Journal of Nutritional Biochemistry, 88, 108532. doi.org/10.1016/j.jnutbio.2020.108532
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Otieno, D., Lee, E., Lee, S., Richard, C., & Kang, H. (2020) Optimizing process of brewing onion peel tea using response surface methodology. NFS Journal, 20, 22-27. doi.org/10.1016/j.nfs.2020.06.004
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Kang, H., Lee, S., Otieno, D., & Ha, K. (2018). Flavonoids, potential bioactive compounds and non-shivering thermogenesis. Nutrients, 10(9), 1168. doi:10.3390/nu10091168
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2020
Citation:
Pei, Y., Kang H. (2020). The effects of quercetin on high fat diet-induced inflammation and oxidative stress in brown adipose tissue. Nutrition 2020
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2020
Citation:
Balogun B., Pei, Y., Kang H. (2020). Garlic scape extract decreases adipogenesis and lipogenesis by activating AMP-activated protein kinase in 3T3-L1 adipocytes
- Type:
Other
Status:
Published
Year Published:
2019
Citation:
Otieno D., Pei Y., Gu I., Lee S., & Kang H. (2019). Effect of quercetin on non-shivering thermogenesis and intestinal microbial populations (P06-037-19). Current Developments in Nutrition, 3 (1), doi.org/10.1093/cdn/nzz031.P06-037-19
- Type:
Other
Status:
Published
Year Published:
2019
Citation:
Otieno D., Altuwaijri S., & Kang H. (2019). In vitro inhibitory effect on carbohydrates digestion and antioxidant potential of garlic scape (P06-076-19). Current Developments in Nutrition, 3 (1), doi.org/10.1093/cdn/nzz031.P06-076-19
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2019
Citation:
Otieno, D., & Kang, H. (2019). Anti-obesity effect of quercetin in high-fat diet-induced obese mice. Proceedings of the 2019 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 10
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2019
Citation:
Altuwaijiri S., & Kang, H. (2019). The effect of in vitro digestion on total phenolic content and antioxidant capacity of garlic scape. Proceedings of the 2019 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 25
- Type:
Other
Status:
Published
Year Published:
2018
Citation:
Otieno, D., Fremah, S., Worku, M., & Kang, H. (2018). Effect of quercetin on non-shivering thermogenesis through Wnt signaling in brown adipocytes (P23-079). Current Developments in Nutrition, 2 (11), doi.org/10.1093/cdn/nzy050
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Richard, C., Otieno, D., Lee, S., & Kang, H. (2018). Optimization of onion peel tea brewing condition using a response surface methodology. Proceedings of the 2018 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 12
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Ha, K., & Kang, H. (2018). Anti-oxidant and anti-hyperglycemia activities of four different colored sweet bell peppers (Capsicum annuum L.) extracts. Proceedings of the 2018 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 13
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Cash, J. O., Ha, K., Otieno, D., & Kang, H. (2018). Anti-inflammatory effects of red onion peel extract on LPS-induced Raw 264.7 macrophage cells. Proceedings of the 2018 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 14
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Otieno, D., Fremah, S., Worku, M., & Kang, H. (2018). Effects of quercetin on non-shivering thermogenesis through Wnt signaling in brown adipocytes. Proceedings of the 2018 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 15
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Progress 10/01/18 to 09/30/19
Outputs
Target Audience:The project is to identify dietary compounds from byproducts of fruits and vegetables that have anti-obesity effect by activating a function of brown adipose tissue and investigate their molecular mechanism. Therefore, target audiences are lab students, scientists who works in nutrient metabolism related to obesity, obese people, and communities who works in food and nutrition such as school teachers and nutritionist in counties. Results and information that are generated through this project were delivered to lab students by laboratory instruction during performing experiments that are proposed in this project. The data was delivered to other scientists and students who works in nutrition and obesity research areas at a scientific conference (annual meeting of American Society for Nutrition, Nutrition 2019) and to school teachers and nutritionist in counties in 2019 Annual meeting of North Carolina Association of Family & Consumer Sciences by poster presentations. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? A new master student received one-on one training about basic experimental skills e.g. gene expression, cell culture and animal handling to assist experiments that are involved in this project. The third year doctoral student had advanced experimental skills to promote her experiments to the advanced metabolic understanding. Also, she is guided to advance onher dissertation writing. Postdoctoral research fellow had advanced training about sophisticated experimental skills and knowledge related to BAT functions and obesity-induced inflammation, which attain greater proficiency to be an independent scientist in molecular nutrition area. How have the results been disseminated to communities of interest?The research has been disseminated to the related and interestedcommunities through several presentations in conferences. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
In objective 1, extract samples from watermelon flesh, skin, and rinds, and garlic skin were prepared. To examine effect of extracts on non-shivering thermogenesis of BAT, cytotoxicity of extracts from watermelon rind and garlic skin, mango kernel, garlic scape, and brewers' spent grain (BSG) were tested in both primary brown adipocytes and 3T3-L1 cells using a MTT assay. Extracts of watermelon rind, flesh, and skin, mango kernel, garlic scape, garlic skin, and BSG at a maximum concentration that did not have cytotoxicity were treated into primary brown adipocytes and TNFa-treated primary brown adipocytes, and garlic scape extract was treated into 3T3-L cells to measure UCP1 gene expression. Extract from garlic skin and BSG reduced UCP1 gene expression whereas garlic scape and watermelon rind extract slightly increased the gene. Mango seed extract did not show any change. 3T3-L1 cells treated with garlic scape extract also slightly showed increased UCP1. However, none of samples resersed TNFa-reduced UCP1 gene expression. In objective 2, to investigate effect of BSG in vivo, mice were fed with low fat-diet (LFD) and high fat-diet (HFD) with or without BSG. Body weight and food consumption were measured weekly. On the completion of the feeding study, glucose and insulin tolerance tests were performed and then blood and tissues were collected. HFD-fed mice supplemented with BSG showed reduced body weight. Glucose clearance rate was not changed by BSG supplementation. To examine effect of quercetin on non-shivering thermogenesis of BAT, the expression of genes and proteins related to non-shivering thermogenesis was analyzed using a real-time PCR and a western blot analysis. Genes that are involved in the regulation of non-shivering thermogenesis including UCP1 were induced. Proteins that are related to AMPK and beta-adrenergic receptor signaling pathways were increased. This data has been delivered to scientists at a scientific conference and publics at NCAFCS conference. A change in knowledge: The results make us be aware of food byproducts that still includes useful dietary compounds that can reduce obesity and their mechanism to regulate obesity. A change in action: Although some extracts showed increased UCP1 gene expression, the data and knowledge that are obtained from this study also lead us to test effect of the extracts on lipid synthesis system to have comprehensive understanding of these extracts-regulated lipid metabolism. Moreover, changes in proteins related to certain signaling pathway led us to improve our project to find a main regulatory system using specific protein-targeting inhibitors or activators. A change in condition: Although stated goals in this project did not meet as the project progresses, activities in this report will eventually contribute to changing people' thought about food byproducts and reducing prevalence of obesity.
Publications
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2019
Citation:
Otieno, D., and Kang, H. (2019). Anti-obesity effect of quercetin in high-fat diet-induced obese mice. Proceedings of the 2019 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 10
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2019
Citation:
Altuwaijiri S., and Kang, H. (2019). The effect of in vitro digestion on total phenolic content and antioxidant capacity of garlic scape. Proceedings of the 2019 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 25
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2019
Citation:
Otieno D., and Kang H. (2019). Quercetin increased non-shivering thermogenesis in brown adipose tissue. Poster session presented at the 2019 ARD (Association of Research Directors) Biennial Research Symposium, 241
- Type:
Other
Status:
Published
Year Published:
2019
Citation:
Otieno, D., Fremah, S., Worku, M., and Kang, H. (2018). Effect of quercetin on non-shivering thermogenesis through Wnt signaling in brown adipocytes (P23-079). Current Developments in Nutrition, 2 (11), nzy050, doi.org/10.1093/cdn/nzy050
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Progress 10/01/17 to 09/30/18
Outputs
Target Audience:This project is a basic-laboratory based project to find dietary compounds from food byproducts that have ability to increase brown fat activity and cause browning effect in white fat, which eventually leads to improving obesity condition by fat loss and further investigate the mechanism by which these dietary compounds regulate non-shivering thermogenesis in brown fat and white fat. Through this project, a master student, a doctoral student, and a postdoctoral research fellow have been trained as target audiences. Knowledge and experimental skills that are related to obesity are delivered to these target audiences by the laboratory instruction. Changes/Problems:One of samples was watermelon rinds. Watermelon's season is summer. To prepare watermelon rinds, we should completely dry them after we sliced them. We used a vacuum ovenfrom another lab (Dr. Yu's lab), but the vacuum did not work after the hurricane. Therefore, we could not dry our sample at that time. All the watermelon rinds that we sliced and prepared were rotten and the summer watermelon season was gone. We will try it again in summer of 2019. What opportunities for training and professional development has the project provided?During the period October 1, 2017-September 30, 2018, one master student, one undergraduate student, one PhD student, and one postdoc have been trained for obtaining new knowledge and experimental skills. Courtney Richard, a URSP student was trained about experimental skills to extract mango seed using organic solvent and measure total phenolic content and antioxidant properties of mango seed's extract and obtained knowledge about antioxidant of mango seed. Olaniyan Jamal Cash, a master student obtained skills to culture RAW macrophage cells and treat the cells using red-onion peel extract. He also obtained experimental skills to measure gene expression from the cells and obtain knowledge about adipocytes related to inflammation in obesity. Dammah Otieno, a PhD student obtained experimental skills to isolate primary brown adipocytes and white adipocytes from brown and white adipose tissue of mice, respectively and culture them. She expanded her knowledge to adipocyte development such as Wnt signaling. Dr. Kyoungsoo Ha worked in the lab shortly. He was trained to extract samples and to perform western blot analysis to measure protein expression, but he was resigned. So, his working period was now enough to contribute to our study. How have the results been disseminated to communities of interest?The dissemination to communities of interest was through a Small Farm Field Day at North Carolina A&T State University. In June 14, 2018, we educated around 100 farmers on health effects and re-usable applications of red and yellow-onion peel and their tea by presenting a poster that was developed for the education purpose regarding "onion peel: reducing obesity and inflammation". Another dissemination to communities of interest was through NCAFCS annual conference by presenting posters about our scientific findings to conference attendances who are usually school teachers and local health related workers and educators. What do you plan to do during the next reporting period to accomplish the goals?In objective 1, we will make extract of garlic skin and watermelon rinds. All samples will be tested in primary brown adipocytes and white adipocytes and then genes will be measured. Based on the concentration of TNF-alpha that we determined, samples will be treated into the TNF-alpha-induced inflamed brown adipocytes and then genes will be measured. In objective 2, mice treated with 1% quercetin will be tested in a metabolic cage to measure energy metabolism and their body temperature and body composition will be measured in Weill Cornell Medicine-metabolic center by a contract service. Diet supplemented with BSG will be designed. Mice will be fed with BSG sample at animal facility of the Wake Forest University.
Impacts
What was accomplished under these goals?
Objective 1 is to examine extracts and dietary compounds of food by-products that have the potential to activate BAT, browning adipocytes, and decrease inflammation in vitro.In this objective, we made extract from mango seeds, garlic scapes, and brewers' spent grain (BSG). We tested cell viability of these extracts using MTT assay in MUCAR cells. We treated one concentration of each extract in primary brown adipocytes and primary white adipocytes to obtain preliminary data. We also tested different concentrations of TNF-alpha in primary brown adipocytes to decide experimental concentration for a further study. We obtained nutrition composition data of BSG sample. Objective 2 is to explore physiological properties of dietary compounds or by-product extracts that have the potential to activate BAT and browning adipocytes e in vivo. In objective 2, we performed a feeding study using 1% quercetin in low and high fat diet fed mice at animal facility of Wake Forest University and collected tissues. The expression of genes related to non-shivering thermogenesis in BAT and WAT were measured.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Kang, H., Lee, S., Otieno, D., & Ha, K. (2018). Flavonoids, potential bioactive compounds and non-shivering thermogenesis. Nutrients, 10(9), 1168. doi:10.3390/nu10091168
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Otieno, D., Fremah, S., Worku, M., & Kang, H. (2018). Effect of quercetin on non-shivering thermogenesis through Wnt signaling in brown adipocytes. Current Developments in Nutrition, 2 (11), 1 November 2019, nzy050, P23-079, doi:10.1093/cdn/nzy050
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Richard, C., Otieno, D., Lee, S., & Kang, H. (2018). Optimization of onion peel tea brewing condition using a response surface methodology. Proceedings of the 2018 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 12
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Ha, K., & Kang, H. (2018). Anti-oxidant and anti-hyperglycemia activities of four different colored sweet bell peppers (Capsicum annuum L.) extracts. Proceedings of the 2018 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 13
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Cash, J. O., Ha, K., Otieno, D., & Kang, H. (2018). Anti-inflammatory effects of red onion peel extract on LPS-induced Raw 264.7 macrophage cells. Proceedings of the 2018 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 14
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2018
Citation:
Otieno, D., Fremah, S., Worku, M., & Kang, H. (2018). Effects of quercetin on non-shivering thermogenesis through Wnt signaling in brown adipocytes. Proceedings of the 2018 NCAFCS (North Carolina Association of Family & Consumer Sciences) Annual Conference, 15
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2017
Citation:
Otieno, D., Parks, J. S, & Kang, H. (2017). Beetroot extract reduced fat accumulation in diet-induced obese mice. IFT (Institute of Food Technologists) 17 Book of Abstracts, 249
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