Progress 12/01/15 to 11/30/19
Outputs
Target Audience:The primary target audience reached during the reporting period is the scientific community. We have published and have submitted abstracts to scientific meetings. Both the Co-PI and PI have made several presentations both in the U.S. and abroad. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?First, the project has supported a post-doc (Christine) and graduate student (Yanling) that would otherwise not exist. Yanling has presented posters at Experimental Biology and participated in summer workshops. She is now a postdoc. Christine is now employed by Agri-Food in Canada. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Aim 1 is completed. We demonstrated that frequent consumption of broccoli alters the microbiome community and function in rats. Specifically, we demonstrated that there was an increase in gut microbial sulforaphane production upon frequent broccoli feeding in rats. More recently, we have preliminary data to show that is also true with kale. Also, we have data with our USDA collaborators that this change in microbiome function also happens in humans fed broccoli or kale. Aim 2 is completed.Although flavonoid sophorosides are common glycosides in brassica vegetables, red raspberries and other food plants, there is a lack of studies of absorption and metabolism of any sophoroside. The aim of this study was to characterize the absorption, phase II metabolism and microbial catabolism of quercetin-3-O-sophoroside, compared to that of quercetin aglycone. Quercetin-3-O-sophoroside was purified from Apocynum venetum and characterized by MS2, 1H and 13C NMR. Using an in situ rat gut model, we found intact, methylated, sulfated and both methylated and sulfated quercetin sophoroside in the plasma following jejunal introduction of the sophoroside; we found derivatives of benzoic acid, phenylacetic acid, and phenyl propionic acid in the cecal contents following cecal introduction. This novel finding, that quercetin sophoroside was absorbed intact, without deglycosylation, points to a possible role for the terminal sugar and/or the type of linkage among glycosidic moieties in the mechanism of absorption of flavonoid glycosides. Aim 3 will continue to be a focus of ourresearch. We have identified a few microorganisms that can consume glucoraphenin (GRP) (>80% consumed in 24 hours) but less than 5% is converted to ITCs. We have been unable to identify the major product of GRP hydrolysis by these bacteria (not SF, erucin, erucin nitrile). One of the candidates was E. coli Nissle. We identified a candidate myrosinase in its genome and made a knockout strain. Yet this strain still consumed GSLs at the same rate despite the hyrdolase knockout. More time is needed to make an exhaustive knockout library of glysosyl hydroases in this strain in order to identify the myrosinase gene. We will continue our work in this area and will be pursuing more funding opportunities.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2019
Citation:
Kaczmarek JL, Liu X, Charron CS, Novotny JA, Jeffery EH, Seifried HE, Ross SA, Miller MJ, Swanson KS and Holscher HD. 2019. Broccoli consumption affects the human gastrointestinal microbiota. Journal of Nutritional Biochemistry. 63. 27-34.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2020
Citation:
Charron CS, Novotny JA, Jeffery EH, Kramer M, Ross SA and Seifried HE. 2020. Consumption of baby kale increased cytochrome P450 1A2 (CYP1A2) activity and influenced bilirubin metabolism in a randomized clinical trial. Journal of Functional Foods. 64.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2020
Citation:
Wang Y, Berhow MA, Black M and Jeffery EH. 2020. A comparison of the absorption and metabolism of the major quercetin in brassica, quercetin-3-O-sophoroside, to that of quercetin aglycone, in rats. Food Chemistry. (In Press).
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Progress 12/01/17 to 11/30/18
Outputs
Target Audience:The primary target audience reached during the reporting period is the scientific community. We have published and have submitted abstracts to scientific meetings. Both the co-PI and PI have made several presentations which presented the ongoing work of the project - both in the U.S. and abroad. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?First, the project has supported a post-doc and graduate student that would otherwise not exist. The graduate student working on the project has presented posters at Experimental Biology and participated in summer workshops. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?Our goals for the next reporting period are tofinalize work on Aim 3 and tofinalize work on Aim 2.
Impacts
What was accomplished under these goals?
Aim 1. We have completed Aims 1.1 and 1.2. The publication from Aims 1.1 and 1.2 has been accepted previously. Aim 1.3 has been completed during this reporting period and is now published. Aim 2. We are still very active with Aim 2 this fall. So far, we have compared gavaging rats with different ratios of glucoraphanin and quercetin. Data analysis of this recently completed animal study is ongoing. Quercetin occurs mostly as quercetin sophorosides in broccoli rather than aglycon. However, little is known about where quercetin sophoroside is hydrolyzed and absorbed along the GI tract (i.e. whether hydrolyzed by lactase-phlorizin hydrolase or cytosolic beta-glucosidase in small intestine or by the microbiota in the lower gut). We have collaborated with Dr. Mark Berhow (USDA, Peoria) for purification of quercetin sophoroside (not commercially available). So far, we have obtained promising results, purifying this from a Chinese herbal tea Apocynum with a promising yield. The purity is >90%. We have studied its metabolism and absorption in the rat model to investigate whether quercetin sophorosides are hydrolyzed by the microbiota in the lower gut, where the hydrolysis of glucoraphanin occurs as well. Manuscript is in development. Aim 3. We have made some progress with Aim 3. We have had significant troubles creating a colorimetric substrate for myrosinase but we have recently changed our strategy. We have identified several candidate microorganisms that we are in the process of confirming. We have identifed a few bacteria that can consume GRP but make an unknown product (not SF, erucin, erucin nitrile). We have also identified a few that can make SF but percent yield is rather low (less than 10%) but far greater than most in the literature. Work in continuing.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Charron, C.S., Vinyard, B.T., Ross, S.A., Seifried, H.E., Jeffery, E.H. and Novotny, J.A. 2018. Absorption and metabolism of isothiocyanates formed from broccoli glucosinolates: Effects of BMI and daily consumption in a randomised clinical trial. British Journal of Nutrition. 120(12): 1370-1379.
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Wang, Y., Jeffery, E.H., Miller, M.J., Wallig, M.A. and Wu, Y. 2018. Lightly cooked broccoli is as effective as raw broccoli in mitigating dextran sulfate sodium induced colitis in mice. Nutrients 10(6).
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Progress 12/01/16 to 11/30/17
Outputs
Target Audience:The primary target audience reached during the reporting period is the scientific community. We have published one manuscript last year. In addition, we have submitted abstracts to scientific meetings. Both the co-PI and PI have made several presentations which presented the ongoing work of the project - both in the U.S. and abroad. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project has supported a post-doc and graduate student. The graduate student working on the project has presented posters at Experimental Biology and participated in summer workshops. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?Our goals for the next reporting period:1. Complete the data analysis described in Aim 1.3 and get it published; 2.Continue our work with quercetin and glucoraphanin regarding the site of absorption and metabolism; and 3.Identify the microbes with significant myrosinase activity.
Impacts
What was accomplished under these goals?
Aim 1. We have completed Aims 1.1 and 1.2. The publication from Aims 1.1 and 1.2 has been accepted. Aim 1.3 is ongoing. We have completed the animal study and are completing final data analysis prior to submitting the manuscript in early 2018. Aim 2. We have become quite active with Aim 2 this fall. So far, we have compared gavaging rats with different ratios of glucoraphanin and quercetin. Data analysis of this recently completed animal study is ongoing. We have recently harvested broccoli with different glucoraphanin and quercetin ratios for use in these future studies. Quercetin occurs mostly as quercetin sophorosides in broccoli rather than aglycon. However, little is known about where quercetin sophoroside is hydrolyzed and absorbed along the GI tract (i.e. whether hydrolyzed bylactase-phlorizin hydrolase or cytosolic beta-glucosidase in small intestine or by the microbiota in the lower gut). We have collaborated with Dr. Mark Berhow (USDA, Peoria) for purification of quercetin sophoroside (not commercially available). So far, we have obtained promising results, purifying this from a Chinese herbal teaApocynum with a promising yield. The purity so far is 40% and we continue working to increase the purity to >90%. Once obtained, we will work on metabolism and absorption in the rat model, to investigate whether quercetin sophorosides are hydrolyzed by the microbiota in the lower gut, where the hydrolysis of glucoraphanin occurs as well. Aim 3. We have made some progress with Aim 3. We have had significant troubles creating a colorimetric substrate for myrosinase but we have recently changed our strategy. We have recently adapted our HPLC method for measuring ITCs to a colimetric process that can be used in a high-throughput manner. We have identified several candidate microorganisms that we are in the process of confirming.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2017
Citation:
Liu, X., Wang, Y., Hoeflinger, J.L., Neme, B.P., Jeffery, E.H. and Miller, M.J. 2017. Dietary Broccoli Alters Rat Cecal Microbiota to Improve Glucoraphanin Hydrolysis to Bioactive Isothiocyanates. Nutrients, 9(3), 262. doi:10.3390/nu9030262.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2017
Citation:
Yanling Wang, Xiaoji Christine Liu, Talon Becker, Se Mi Song, John A Juvik, Michael J Miller and Elizabeth H Jeffery. 2017. Abstract: The Role of the Gut Microbiome in Quercetin Inhibition of Sulforaphane Bioavailability and Bioactivity in Rats. The FASEB Journal vol. 31 no. 1 Supplement 646.4.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2017
Citation:
Jeffery, E.H. 2017. Abstract: Controlling Obesity-Derived Hepatic Lipidosis and Carcinogenesis Through Dietary Broccoli. 15th World Congress on Advances in Nutrition, Food Science & Technology. Edinburgh, Scotland Sept 11-12, 2017.
- Type:
Other
Status:
Other
Year Published:
2017
Citation:
Jeffery, e.h. 2017. Comparing Cooked and Raw Broccoli Efficacy for Colon Health.
Glucosinolate Conference 2017. Berlin Germany Sept 17-20, 2017.
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Progress 12/01/15 to 11/30/16
Outputs
Target Audience:The primary target audience reached during the reporting period is the scientific community. We have submitted one manuscript that is currently under review. In addition, we have submitted two abstracts to scientific meetings. Both the Co-PI and PI have made several presentations which presented the ongoing work of the project - both in the US and abroad. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?First, the project has supported a post doc and a graduate student that would otherwise not exist. The graduate student working on this project presented a poster at Experimental Biology (see Products section for more details). How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?Our goals for the next reporting period: 1. Get our publication focused on Aim 1.1 and 1.2 published. 2. Complete large animal study for Aim 1.3 and anlayze/intrepret the data. We should also have this work submitted for publication by the end of the next reporting period. 3. Initiate quercetin/sulforaphane whole broccoli animal studies. 4. Make progress in identifying the major source of microbial myrosinase in the rat cecum.
Impacts
What was accomplished under these goals?
Our group and others have demonstrated many health-promoting benefits of consuming whole broccoli. However, the impact of whole broccoli on gut health is largely unknown. As an outcome of the proposed investigations, we will have determined that whole broccoli consumption alters the gut microbiota composition and metabolism. These alterations of gut microbiota composition and metabolism are likely to increase microbial SF production, gut SF absorption and gut SF bioactivity. Together, we expect a positive contribution to gut health and a corresponding reduction in inflammation. Lastly, the identification of microbial myrosinases will help explain the variation in clinical response to GRP and thus enable novel approaches to improve microbial SF production in the gut. These expected outcomes are significant, because they can be used to inform the public about the intestinal health promoting properties of whole broccoli. Information about any synergies of Quercetin with SF can inform plant breeders for improved selection of new cultivars. In addition, identification of a microbial myrosinase will enable strategies to increase microbial SF production in the gut, so that consumers that prefer cooked broccoli can still gain the health benefits. Furthermore, understanding much of the past variation in response to GRP will allow more uniform clinical studies potentially leading to establishment of health claims. Aim 1. We have completed Aims 1.1 and 1.2. The publication from Aims 1.1 and 1.2 is currently under review. Aim 1.3 is ongoing. We have recently completed a small pilot study to establish the basic parameters of the DSS-induced inflammation rat model. We expect to make significant progress with Aim 1.3 during year 2. Aim 2. We have become quite active with Aim 2 this fall. So far, we have compared gavaging rats with different ratios of glucoraphanin and quercetin. Data analysis of this recently completed animal study is ongoing. During spring 2017, we anticipate using whole broccoli to test our hypothesis. Towards that end, we have recently harvested broccoli with different glucoraphanin and quercetin ratios for use in these future studies. Aim 3. We have made some progress with Aim 3. We have had significant troubles creating a colorimetric substrate for myrosinase but we have recently started collaborating with a carbohydrate chemist that is working on the problem. We have also attempted to use glucoraphanin and/or sulforaphane to select for myrosinase postive microorganisms from our rat studies. We have identified several microorganisms that can convert glucoraphenin to sulforaphane. We are in the process of further evaluations of these microbes.
Publications
- Type:
Other
Status:
Accepted
Year Published:
2016
Citation:
Effect of frequent broccoli intake on rat gut microbiota metabolism and composition. Experimental Biology. San Diego, CA, Apr 2-6, 2016 - Poster.
- Type:
Other
Status:
Accepted
Year Published:
2016
Citation:
The role of the microbiome in bioavailability of sulforaphane from dietary broccoli. Food Bioactives and Health Conference. Norwich, UK, Sept 13-15, 2016 - Poster.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2016
Citation:
The role of the microbiome in bioavailability of sulforaphane from dietary broccoli. Food Bioactives and Health Conference. Norwich, UK, Sept 13-15, 2016 - Oral Presentation.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2016
Citation:
A new era for prebiotics: Improving activation of bioactive food components by the microbiome. Food Technology Conference. Las Vegas, NV. November 14-16. Keynote Speaker.
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