Progress 07/01/18 to 06/30/19
Target Audience:Dr. Seeram and Dr. Slitt presented their results on this grant at the American Chemical Society national meeting in Spring 2020 in Orlando to over 50 attendees. Also, Dr. Seeram presented data as the Keyonote speaker at the annual meeting of t the International Maple Syrup Institute in October 2020 inMinnesota. Changes/Problems:
What opportunities for training and professional development has the project provided?Several undergraudate students worked on the grant. How have the results been disseminated to communities of interest?To themaple growers and producers at the natinonal meeting of the IMSI. Also,on the website www.uri.edu/maple What do you plan to do during the next reporting period to accomplish the goals?Plan to publish a manuscript
What was accomplished under these goals?
Background: Obesity is a diet-induced disease that stems into numerous pathologies such as metabolic syndrome, type II diabetes mellitus, Nonalcoholic Steatohepatitis, Fatty Liver disease and even cancer. Chronic low grade inflammation is linked in the etiology of these diseases. Polyphenol rich nutraceuticals such as Maple Syrup Extract (MSX) may prevent or reduce the severity of these inflammatory processes. Methods: MSX was evaluated in a murine diet-induced obesity model for a period of 18 weeks. Body weight and food consumption were monitored in addition to measuring Fasting Blood Glucose, Glucose, and Insulin Tolerance during this study. IL-6 was quantified from serum following necropsy. Liver and White Adipose Tissue were excised and weighed at the time of necropsy. Livers were sectioned, stained using H&E and scored for histological features associated with NASH/NAFLD. Livers were also subjected to bottom-up proteomic analysis for significant protein changes. Results: 18 weeks of high fat high fructose consumption paired with sedentary behavior caused significant increases in body weight, serum IL-6 concentrations, glucose and insulin intolerance as compared to low fat diet fed mice. The addition of MSX 0.5% w/w exacerbated liver steatosis whereas MSX 0.05% w/w failed to improve histological scoring as compared to high fat diet control mice. Conclusion: MSX supplementation in high fat diet feed did not ameliorate the progression of fatty liver disease in this rodent model as compared to our previous pilot study. The addition of MSX at 0.05% did significantly reduce pro-inflammatory serum IL-6 concentrations as compared to high fat diet control mice. The addition of fructose and glucose in drinking water combined with the duration of diet exposure may have masked the beneficial effects of MSX witnessed in previous studies.
Conference Papers and Presentations
1. Maple-derived food bioactives in human health & diseases: The path forward. Seeram N.P. American Chemical Society (ACS) National Meeting, Orlando, FL, March 30, 2020. (Oral).
Progress 07/01/17 to 06/30/18
Target Audience:We have had two principal audiences - scientists and the public. We had communicated our pilot animal studyfindings on MSXat the annual American Chemical Society meeting in April 2017 at the First Global Symposium on Chemistry and Biological Effects of Maple Food Products in the Division of Agricultural and Food Chemistry. Dr. Seeram (co-I) was the opening speaker at the International Polyphenol Congress held in Madison WI in July 2018 and he presented data on maple polyphenols incluiding MSX. Dr. Slitt (Co-I) also presented our MSX work at a local AAPS meeting in May 2018 and at the USDA Project Directors Meeting in Boston 2018. Dr. Seeram has also done multiple public speaking presentations at several RI Garden Clubs on maple phytochemicals and MSXincludingNewport Garden Club (November 2017), South Kingston Garden Club (September 2018)and Arbutus Garden Club in Charlestown (October 2018). Changes/Problems:The small deviations for the project were that we selected a slightly different diet than proposed. There were some concerns from the reviewers that we needed to have a lower percent fat diet. Our initial study used a 45% kCal diet, but this diet did not significantly induce inflammation. Therefore, we decided to switch to a American Life Style Obesity (ALIOS) diet, which is a western diet (22% hydrogenated vegetable oil, 0.5% cholesterol) along with high fructose-sucrose water (42g/L) to induce more inflammation and a non-alcoholic steatosis phenotype. We also adjusted the timing of our tissue collections, with the first collection being at week 18 instead of week 15 and our second time point at week 34. Basically, we measurements in body weight, glucose sensitivity, and insulin sensitivityto better gauge the appropriate timing. A second addition to the project was to include THP-1 human monocytes in addition to the primary monocytes proposed. A major unanticipated finding is that high concentrations of MSX in feed or culture media exert adverse effects. We have observed that lower MSX concentrations (1 ppm) that are more relevant to consumption exert much more significant anti-inflammatory properties than high concentrations (100 ppm). This suggests that there might be some constituents of MSX that have toxic properties, but only at high, non-relevant concentrations. What opportunities for training and professional development has the project provided?The project has funded 1 graduate research assistant. The graduate assistant (Nick DaSilva) has received an interdisciplinary training in medicinal chemistry and pharmacology/toxicology. The graduate student has participated in routine lab meetings in Dr. Seeram and Dr. Slitt's research groups. He has received training in culture of human monocytes, as well as conducting a large-scale rodent experiment. Since the start of this study, our group has trained approximately 4undergraduate students per semester who were exposed to a number techniques and skillsets such as basic cell culture, animal welfare/feeding, and experiment design/execution/data analysis. This study has also been used to initiate collaborations with several research groups (domestic and foreign) to further investigate the effects of MSX in various tissues from this rodent study namelykidney, small intestines and fecal microbial diversity. How have the results been disseminated to communities of interest?Results have been disseminated primarly through professional research societies such as the American Chemical Society and the American Society of Nutrition (ASN). What do you plan to do during the next reporting period to accomplish the goals?The final necropsy is to be performed withtissues harvested and analyzed as described in previous sections. Specifically, much of the tissue analysis that remains is to be performed by collaborating research groups on a contractualbasis. These results will then be compiledin manuscript form for submission into a suitable journal within the grant specified timeline (June 2019). We are currently preparing at least two resulting manuscripts from the funded work and will present the findings at the Spring ACS meeting in Orlando, Florida in a symposiumentitled " Global Symposium on Chemistry and Biological Effects ofMaple-Food Products". This session is being organized by a member of the PI's group (Dr. Hang Ma) and the PI (Seeram) and Co-I (Slitt) will be presenting research findings at the symposium. The PIs have also forged collaborations with other experts to expand some of the proposed endpoints to areas that are emerging and novel to Maple. For example, Dr. Seeram's group has collected fecal material from the funded mouse study and the impact of MSX feeding on gut microial diversity will be assessed by16S rRNA sequencing performed by Dr. Andree Marette's group at University Laval in Canada. Kidney samples from the resulting study will be analyzed by Dr. Nathan Cherrington's research group at the University of Arizona. Lastly, through collaborations at the Universit of Rhode Island, our group as acquired expertise in label-free proteomics methodology by MS-SWATH for untargeted proteomics.
What was accomplished under these goals?
We are about 75% completed performing the proposed studies for pre-adipocyte work with murine 3T3-L1 adipocyes, with some timepoints and modifications to the culture conditions remaining. We have observed that treatment with maple syrup extract (MSX) can slightly decrease lipid accumulation and decreases LPS-induced cytokine secretion into media. We have completed the proposed human monocyte experiments and were able to add an additional human monocyte/macrophage cell line. This work was able to demonstrate that LPS can stimulate Tnf-alpha and Il-6 secretion into cell culture media and this was significantly decreased by co-treatment with MSX (0.1 ppm and 1 ppm). We have completed week 17 of our in vivo mouse study and are currently on the week 30timepoint. We are getting ready to necropsy the mice. We have also measured and asessedfasting blood glucose,glucose and insulin tolerance. Liver samples will be examined and scored by a trained pathologist.