Progress 01/01/24 to 12/31/24
Outputs
Target Audience:Ulcerative colitis is a disease experienced worldwide. Understanding to role that the complex (polyphenols and polysaccharides) can play in alleviating the symptoms will be of interest to a variety of individuals, including undergraduate and graduate students, scientists, practitioners and individuals suffering from Ulcerative Colitis. The results werepresented to undergraduate and graduate students, scientists, practitioners and industry personnel. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Downstream experiments have trained students on techniques including mammalian cell culture, LC-MS, RT-PCR, GC-MS, DNA extraction, bacterial cell culture and proteomic extraction. In addition, graduate students are learning how to breed mice and how to conduct a study using mice as a model organism through hands-on training and animal care. The project also helped train undergraduate students in gut health research. How have the results been disseminated to communities of interest?The results were presented at departmental seminars and national conferences. Presentations were presented as part of the Whistler Center Update Series and posters were presented at the 2024 Whistler Center Industrial Associates Meetings. Further, an abstract and poster was presented at the 2025 Crohn's and Colitis Congress and 2024 Institute of Food Technologists Expo. What do you plan to do during the next reporting period to accomplish the goals?Cecal digesta is currently being sequenced to determine if supplementation of the complex alters the gut microbiota in mice that have undergone DSS treatment. In addition, colon tissues from mice in the therapeutic study are currently being analyzed for inflammation and oxidative stress. Lastly, Ulcerative colitis associated bacteria will be transplanted into germ-free IL-10 knockout mice to determine how the complex can alter the gut microbiota. Another study utilizing a bioengineered Lactobacillus probiotic for increased adhesion will be conducted in conjunction with blueberry polyphenols in an IL-10KO/ Piroxicam model to assess if a stronger Lactobacillus community may be able to metabolize the blueberry polyphenols more efficiently and confer resulting health benefits. The findings will be disseminated at national conferences and meetings.
Impacts
What was accomplished under these goals?
Ulcerative colitis (UC) is characterized by chronic inflammation in the colon, oxidative stress, and bacterial dysbiosis. Anthocyanins are known for their health-promoting properties, including anti-inflammatory and antioxidant effects, modulation of gut microbiota, and production of beneficial byproducts such as short-chain fatty acids when metabolized in the colon. These effects are most pronounced when anthocyanins reach the gut microbiota in the colon. However, isolated anthocyanins are unstable in the digestive tract due to pH fluctuations and high temperature, resulting in low colonic concentrations. Anthocyanin and pectin complexes increase the stability of anthocyanins, which was reported in a previous year's progress report, and can potentially encourage the delivery of active anthocyanins into the diseased area, reducing overall inflammation and oxidative stress while also being a beneficial compound for gut bacterial metabolism. We have determined that the anthocyanin and pectin complex can form not only when the anthocyanins are isolated but also within the whole food matrix. We successfully enhanced the interactions between blueberry anthocyanins and additional pectin within a whole food matrix. The resulting freeze-dried blueberry powder exhibited improved anthocyanin stability and bioactivity following in vitro upper GI digestion. To evaluate the treatment potential of this complex, we used IL-10 knockout mice, a model for UC. The complex was incorporated into mouse chow and fed to the mice. The protective effects of the complex are being tested using a DSS-induced colitis model. Preliminary results show that the complex prevents colon shortening (a hallmark of DSS-induced UC), significantly reduces inflammatory gene expression, increases tight junction RNA expression, and lowers oxidative stress. Gut microbiota composition is currently being analyzed via 16S rRNA sequencing. In addition, the complex is being evaluated in a second UC model induced by piroxicam. Early findings suggest that supplementation after disease induction leads to faster recovery and weight gain. Tissue samples from this study are currently being analyzed for inflammatory and oxidative stress markers. Both probiotics and whole foods rich in polyphenols aredietary interventions thathave been explored individually to help manage and treat IBD with great success. However, the ideal combination of both compounds leaves much to be desired. Thus, these compounds were tested together in vitro and in vivo. Currently, the in vitro experiment is completed with analysis of total phenolics, and antioxidant ability assessed with co-incubation of Lactobacillus species and blueberry polyphenols. The efficacy of polyphenols increasing adhesive capability in a colon epithelial cell line has also been determined. In vivo study preliminary results indicate that supplementation of blueberry polyphenols was able to increase tight junction protein expression and reduce inflammation more profoundly than the combination of the compound with probiotics. Microbiome analysis from the treatment groups is currently underway.
Publications
- Type:
Peer Reviewed Journal Articles
Status:
Published
Year Published:
2024
Citation:
Nathan V. B., Eckrote S., Li S., & Reddivari L. 2024. Crude Blueberry Phenolic Extracts Improve Gut Barrier Integrity and Exert Anti-Inflammatory and Antimicrobial Activity in an In Vitro Weaning Stress Model. Antioxidants, 13(9), 1044.
- Type:
Peer Reviewed Journal Articles
Status:
Published
Year Published:
2024
Citation:
Moncada, E., Bulut, N., Li, S., Johnson, T., Hamaker, B., & Reddivari, L. (2024). Dietary Fibers Physicochemical Properties and Gut Bacterial Dysbiosis Determine Fiber Metabolism in the Gut. Nutrients, 16(15), 2446.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Nathan, V (2024). Effect of Blueberry Polyphenols on Improving Intestinal Health of Weaning Piglets. Institute of Food Technologists Expo, July 14-17, Chicago, IL.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Moncada, E., Andino, P., Johnson, T., Hamaker, B., & Reddivari, L. (2024). Dietary Fiber Structure Complexity Influences Fiber-Tolerance in a Humanized Germ-Free IL-10 KO Mice. Institute of Food Technologists FIRST. July 14-17, Chicago, IL.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Moncada, E., Andino, P., Johnson, T., Hamaker, B., & Reddivari, L. (2024). Exploring the Role of Gut Bacterial Dysbiosis and Gut Inflammation in Fiber Tolerance. American Society of Nutrition, Curr Develop Nutr, Volume 8; Supplement 2.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Moncada, E., Andino, P., Johnson, T., Hamaker, B., & Reddivari, L. (2024). Deciphering Fiber Tolerance: Impact of Gut Bacterial Dysbiosis and Fiber Physicochemical Properties. Midwest Microbiome Symposium. May 13-15, West Lafayette, IN.
|
Progress 01/01/23 to 12/31/23
Outputs
Target Audience:By completing the objectives, the team aims to inform scientists and practitioners in the agriculture industry on the merits of using whole food supplements as potential feed additive in swine care. Also, scientists, practitioners and individuals suffering from Ulcerative Colitis can be informed of the role that anthocyanin and pectin complexes have in alleviating Ulcerative Colitis symptoms. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?One Master's student who graduated working on part of this project is now pursuing PhD. The project has allowed graduate students to learn how to conduct a study utilizing swine as a model organism as they received hands-on training with swine care. Downstream experiments have trained students on techniques such as mammalian cell culture, LC-MS, RT-PCR, DNA extraction, and proteomic extraction. In addition, graduate students are learning how to breed mice and how to conduct a study using mice as a model organism through hands-on training and animal care. The project also helped train undergraduate students in gut health research, who are pursuing grad school now. How have the results been disseminated to communities of interest?The results were presented at departmental seminars and national conferences. Posters were presented and abstracts were published as part of 2023 American Society for Nutrition Conference. Posters will be presented at Whistler Center Industrial Associates Meetings. The poster abstract was also accepted for presentation at the Institute of Food Technologists' annual meeting in July 2024. What do you plan to do during the next reporting period to accomplish the goals?Piglet distal colon samples have been sent for proteomic analysis to further corroborate the hypothesis that antioxidant pathways were altered. Fecal samples from the piglets are currently being sequenced to determine if supplementation altered the piglet microbiome composition. Complete mice studies investigating the impacts of anthocyanin and pectin complex on inflammation and oxidative stress using genetic and DSS-induced chronic colitis models. Disseminate the findings at national conferences.
Impacts
What was accomplished under these goals?
Objective 1 and 2a: Ulcerative Colitis is associated with significant amounts of inflammation in the colon, oxidative stress, and bacterial dysbiosis. Anthocyanins have a variety of health-promoting properties including anti-inflammatory, antioxidant, positive gut bacterial interactions, and beneficial byproduct production when metabolized by gut microbiota. These benefits are more pronounced when the anthocyanins can reach the gut microbiota in the colon. Isolated anthocyanins lack stability in the digestive tract because of the changes in pH and colonic concentrations of anthocyanins are low. Anthocyanin and pectin complexes increase the stability of anthocyanins, which was reported in last year's progress report, and can potentially encourage the movement of the active anthocyanins into the diseased area, reducing overall inflammation and oxidative stress while also being a beneficial compound for gut bacterial metabolism, producing short chain fatty acids. The ability of anthocyanin and pectin complexes to aid in alleviating symptoms associated with Ulcerative Colitis is being determined using IL 10 knockout mice, an Ulcerative Colitis model. We successfully enhanced the interactions between blueberry anthocyanins and pectin in situ. The resulting freeze-dried blueberry powder showed increased anthocyanin stability. The blueberry anthocyanin and pectin complex was incorporated into chow and is currently being fed to the knockout mice. The current study is investigating the protective ability of the complex on DSS-induced Ulcerative Colitis. In addition, the therapeutic potential of the complex will be determined through the induction of Ulcerative Colitis in the IL 10 knockout mice. Both studies are underway. Weaning is considered one of the most stressful events in a piglet life due to debilitated gastrointestinal function, increased inflammation, and immense oxidative stress. Polyphenols and anthocyanins are bioactive compounds that have shown merit in mitigating these symptoms thus these compounds were tested in vitro and in vivo. Currently, the in vitro experiment is completed and is awaiting results to determine the composition of the crude blueberry polyphenolic extract through metabolomic profiling of anthocyanins present. In vivo study: 20 early weaning piglets were supplemented with 10% freeze-dried blueberry powder in addition to their basal diets for three weeks. Preliminary results indicate that supplementation of these compounds did not significantly alter tight junction protein expression or reduce inflammation in the early weaned piglet model. However, the antioxidant status of blueberry-treated piglets was significant. Antioxidant enzymes, catalase and superoxide dismutase were upregulated. Furthermore, thiobarbituric acid, a marker of lipid peroxidation, was markedly decreased in treated piglets.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Fu W, Li S, Helmick H, Hamaker BR, Kokini JL, Reddivari L. Complexation with Polysaccharides Enhances the Stability of Isolated Anthocyanins. Foods. 2023; 12(9):1846. https://doi.org/10.3390/foods12091846
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Moncada E, Bulut N, Li S, Hamaker B, Reddivari L, SELECTION OF DIETARY FIBERS FOR GUT HEALTH BASED ON THEIR PHYSICOCHEMICAL PROPERTIES, Inflammatory Bowel Diseases, Volume 29, Issue Supplement_1, February 2023, Page S48
- Type:
Journal Articles
Status:
Under Review
Year Published:
2023
Citation:
Moncada E, Bulut N, Li S, Johnson T, Hamaker BR, Reddivari L*. 2023. Selection of Dietary Fibers for Gut Health based on their Physicochemical Properties: In-vitro study In review Food and Function
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Nathan V, Eckrote S, Li S, Reddivari L. Effect of Crude Blueberry Phenolic Extracts on Improving Gut Barrier Integrity and Inflammation In Vitro. American Society of Nutrition, July 22-25, 2023 Boston, MA Current Developments in Nutrition Issue 7, Supplemental 1, pg 206
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Reddivari L, Moncada E, Li S. OR16-05-23. Role of Fiber Physicochemical Properties in Fiber Intolerance, Current Developments in Nutrition, Volume 7, Supplement 1, 2023, 100837, ISSN 2475-2991
|
Progress 01/01/22 to 12/31/22
Outputs
Target Audience:Scientists, Graduate and Undergraduate students, Industry personnel and Consumers Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Provided an opportunity to interact with other scientists as part of W40002 and to train undergraduate students, graduate students and Post-doc in the area of plant bioactive compounds and their health benefits. This grant helped an undergraduate student to be accepted into the Purdue Summer College of Agriculture Research Fellowship (SCARF) and receive the Office of Undergraduate Research Grant. How have the results been disseminated to communities of interest?The results were presented in seminars and symposiums. The attendees represent scientists, industry personnel, graduate and undergraduate students in nutrition, food science, chemistry and plant science. We published three abstracts. What do you plan to do during the next reporting period to accomplish the goals?We are breeding germ-free IL-10 KO mice and established the model for the colonization of Ulcerative colitis-associated bacteria.We will conduct animal experiments using both germ-free mice colonized with healthy and colitis-associated gut bacteria to understand the role of microbiota dysbiosis in the anti-colitic potential of Blueberry and pectin complex.
Impacts
What was accomplished under these goals?
Though the grant start date was January 2022, funds were received in May 2022. Research Hypothesis: Phenolic Compounds from blueberries will mitigate the adhesion and invasion of Enterotoxigenic Escherichia coli (ETEC) and Salmonella enterica Serovar Typhimurium (ST) to porcine intestinal epithelial cells. Weaning is a stressful season in a piglet life and marks the transition to an adult diet. The process is accompanied by impaired gastrointestinal tract (GIT) function resulting in post-weaning diarrhea (PWD). PWD leads to an immunocompromised piglet, thus making the piglet more susceptible to bacterial infections. Currently, antibiotics are used as a precautionary measure, but the concern of antibiotic resistance in veterinary medicine has increased the need for a nutraceutical approach. Anthocyanins are bioactive compounds found in a variety of fruits and vegetables and are proven to improve gut health as well as contain antimicrobial properties. Here, we investigated the prophylactic effects of commercially available blueberry phenolic compounds (BPE), rich in anthocyanins, on IPEC-J2 cells infected with Enterotoxigenic Escherichia coli (ETEC) and Salmonella enterica Serovar Typhimurium (ST). Minimum inhibitory concentrations for ETEC and ST were 250ug/mL and 62.5ug/mL, respectively. BPE did not significantly decrease the adhesion or invasion of either pathogen. However, the antioxidant status and cell stress will be assessed to determine if the inflammatory response is reduced. The preliminary data suggests that BPE did not mitigate the adhesion or invasion of ST and ETEC. Research Hypothesis: The stability and bioavailability of anthocyanins can be improved with polysaccharide complexation Isolated anthocyanins are unstable and can be easily destabilized by environmental factors (pH, light, temperature). Furthermore, the colonic concentration of isolated anthocyanins is low compared to when in a food matrix. Complexation is a technique used to stabilize anthocyanins. This study aims to investigate which polysaccharides can stabilize anthocyanins and enhance the colonic concentration of anthocyanins via complexation. The health benefits of anthocyanin-polysaccharide complexes were also evaluated in vivo. The different polysaccharides used were pectin, inulin, starch, cellulose, and their mixture in equal proportions. Pectin exhibited the best binding capacity with anthocyanins (anthocyanin-pectin complex; APC). APC retained more anthocyanins compared to other complexes after the simulated upper GI digestion. The electrostatic interaction of APC quantified by zeta potential showed that anthocyanins were positively charged, and pectin was slightly negatively charged under pH 2.0 conditions. The increased particle size of APC was observed indicating the agglomeration between particles. The physical surface structure of APC was changed after complexation compared to APM (anthocyanin-pectin mixture) as observed by SEM. The stability of anthocyanins in APC was better at pH 3, 5 and 7 and showed higher temperature tolerance than isolated anthocyanins. Furthermore, APC significantly decreased the LPS-induced increase in gut permeability in Caco-2 cell model. APC restored the gut barrier function in vivo by reducing gut permeability and increasing mucus thickness in a DSS-induced colitis mouse model. APC also increased the total short-chain fatty acids, especially butyrate concentration. In conclusion, APC enhanced the stability of isolated anthocyanins and improved the gut barrier function compared to isolated anthocyanins.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Eckrote S, Nathan V, Reddivari L. 2022. Protective Effects of Blueberry Phenolic Compounds in IPEC-J2 Cells Challenged with Post Weaning Diarrhea Pathogens. Fall Undergraduate Research Expo, Purdue University Poster Competition.
- Type:
Other
Status:
Other
Year Published:
2022
Citation:
Nathan V, Reddivari L. 2022 Effect of Crude Blueberry Phenolic Extracts on Improving Gut Barrier Integrity and Inflammation in IPEC-J2 Cells Challenged with LPS. Whistler Center for Carbohydrate Research Progress Update, Purdue University- Research Talk
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Eckrote S, Nathan V, Reddivari L. 2022. Anti-pathogenic Activity of Blueberry Phenolic Compounds on Salmonella enterica Serovar Typhimurium (ST) and Enterotoxigenic E. Coli (ETEC) in IPEC-J2 Cells. Summer Undergraduate Research Symposium, Purdue University Poster Competition.
|
|