Progress 10/01/10 to 09/30/14
Outputs
Target Audience: The target audiences of this project were scientists, health professionals, animal nutritionists, farmers and dairy producers. Information gathered as a result of this project has been disseminated among the target audiences through peer-reviewed publication, seminar presentations, consultation with dairy producers, and collaboration with laboratories in similar research areas. Changes/Problems: The transitioning of graduate students (Shane Huebner and Jake Olson) slowed the progress of this project as the new graduate student Jake Olson required training prior to conducting experiments. A co-PI on the project (Fariba Assadi-Porter), left the University of Wisconsin-Madison for another position, which slowed the progress of goals 1 and 3 as she was responsible for teaching the metabolomics analysis methodology to Jake Olson and the facility to conduct experiments thereof. Discovery that c9t11-CLA was as effective as the recommended human dose of Celebrex (celecoxib) made objective 4 obsolete (this objective was to determine if c9t11-CLA could serve as an adjunct to reduce levels of Celebrex needed to treat inflammation). Hence focus of the research continues to study metabolomic dysfunction, objectives which are ongoing and will be completed in 2015. What opportunities for training and professional development has the project provided? One graduate student (Jake Olson) was trained by the graduate student (Shane Huebner) in laboratory practices integral to the completion of the project goals. Professor Lai Changhua, College of Animal Science and Technology, China Agriculture Beijing, provide critical metabolomics input during the project. Additionally, several undergraduate students have been trained and mentored by Jake Olson listed as follows: 1) Alexander Haas, 2) Holly McKee, 3) Jennifer Lor, 4) Joni Baker, 5) Jennifer Roberts, 6) John Brantingham, and 7) Sarah DeVitt. Two undergraduate students, Joni Baker and Holly McKee presented mentored research findings at the yearly Introductory Biology 152 Undergraduate Research Poster Presentation at UW-Madison. Two graduate students, Daniel Zwick and Nitasha Bennet, have also been trained in animal husbandry techniques associated with this project by Jake Olson. Since the onset of this project, Shane Huebner has given one seminar presentation for the Nutritional Sciences seminar series and graduated with a Ph.D. at the end of the first year of this project (2011). Jake Olson joined the project after the departure of Shane Huebner and has given three seminar presentations regarding the progress and findings of this project for the Molecular and Environment Toxicology seminar series (2012-2014). How have the results been disseminated to communities of interest? Results of this project have been disseminated to communities of interest through peer-reviewed publication, outreach activities that include consulting with Wisconsin dairy producers and the analytical testing of dairy products for CLA content. Cook co-organized the 2011 Kenneth A. Spencer Award Symposia at the 244th American Chemical Society meeting entitled "Conjugated linoleic acid (CLA): A naturally occurring anti-obesity, anti-inflammatory fatty acid." The symposia included 8 invited speakers from around the world who are considered experts on CLA. Cook and lab members also presented invited talks at Ohio State University, University of Chicago, British Society of Animal Sciences in Nottingham, England, American Oil Chemist Society in Cincinnati, Ohio, the 9th Mid-Atlantic Nutrition Conference in Baltimore, Maryland, and at the American Chemical Society-Kansas City Spencer Award Symposia in Kansas City, MO. An Internal Food Technologist webinar entitled, "Conjugated linoleic acid from discovery to GRAS, a 20 year journey" was produced and made available nationally. Seminar presentations at the Molecular and Environmental Toxicology seminar series has allowed the results of this project to be disseminated to researchers of various disciplines and broad research areas. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
As chronic inflammation and diseases associated with persistent inflammatory activity are increasing in the US, an overall goal of this project was to determine whether the intake of ruminant derived dietary fat, cis-9, trans-11 conjugated linoleic acid (c9t11-CLA), can reduce inflammation as well as or more effectively than a prescription non steroidal anti-inflammatory (NSAID) over time. Experiments were designed to specifically compare the NSAID, celecoxib, the most prescribed NSAID in the US, against c9t11-CLA for anti-inflammatory effects in animals. Previously, our lab has shown that c9t11-CLA is effective at reducing arthritic severity greater than 30% in arthritic mice at a concentration as low as 0.5% of the total diet. Since this dietary fat is a natural product of grass-fed ruminants and highly abundant for potential mass consumption by humans our research led us to examine c9t11-CLA's anti-inflammatory effects compared to celecoxib. Celecoxib has been shown to impart several side effects including heart attack, stroke, and thrombosis, leading us to hypothesize that this drug may cause long-term aberrations in normal metabolism, ultimately leading to comorbidity when taken over long periods of time. We then sought to examine the effects of c9t11-CLA on metabolism, comparing any potential changes to arthritic animals treated with celecoxib as a method of both comparison and potentially predicting side effects of long-term use. Finally, since it was unknown whether or not c9t11-CLA would impart similar anti-inflammatory activity as celecoxib, an experiment was proposed to examine the interaction of c9t11-CLA and celecoxib on inflammation in a combined treatment. Two graduate students, Shane Huebner and Jake Olson, have examined blood plasma from arthritic animals treated with c9t11-CLA, celecoxib, or corn oil (control) for alterations in small molecule metabolites as a result of long-term (84 days) dietary treatment. Our findings indicate that chronic inflammation significantly decreases total plasma amino acids (38% decrease) in arthritic mice and this effect was minimized in arthritic mice fed 0.5% c9t11-CLA of diet (20% decrease) to the extent that c9t11-CLA treated mice had total amino acid concentrations statistically indifferent from healthy non-arthritic mice. Jake Olson has performed an experiment to establish the dietary equivalent of c9t11-CLA to celecoxib for the reduction of inflammation in mice over a 42-day treatment period. Data generated from this study are currently being analyzed and interpreted; however, it is clear from our results that 0.5% dietary c9t11-CLA is at least as effective as 5mg/kg (relevant human daily dose) celecoxib at significantly reducing inflammation by 42 days post treatment. An inflammatory metabolomic analysis is currently being conducted to identify any changes in metabolism due to treatment. 1. Conduct an inflammatory metabolomic analysis of plasma samples from previous experiments. An experiment was conducted to profile metabolic changes due to dietary treatment in plasma from both non-arthritic and arthritic mice. After 42 days of treatment with either 1% corn oil or 0.5% c9t11-CLA + 0.5% corn oil, blood was collected from mice. Low molecular weight metabolites were purified and analyzed using NMR spectroscopy. Metabolite concentrations were profiled using the computer software Chenomx and analyzed for differences due to treatment. A major finding from this experiment was that arthritic mice regardless of treatment had decreased plasma amino acid concentrations; however, CLA treated animals had a decreased arthritic severity compared to corn oil treated arthritic mice as well as a decreased reduction in amino acid concentration, such that CLA treated mice were indifferent from non-arthritic mice. These findings suggest chronic inflammation perturbs systemic metabolism toward altered utilization of amino acids and that a decrease in plasma amino acid content may be a useful biomarker of chronic inflammation. Additionally, these results suggest that dietary CLA, in addition to being an effective treatment for reducing inflammation in animals, may also be useful in restoring an inflammation-linked, perturbed amino acid metabolome to normalcy. 2. Determine the "COX-2 equivalency" for c9, t11-CLA. An experiment was conducted by feeding three separate doses of the NSAID celecoxib (0.5, 5, and 50 mg/kg bw) to arthritic mice over a 42-day period, as well as a dietary treatment of 0.5% c9t11-CLA to determine a dose of celecoxib equivalent to c9t11-CLA for reducing inflammation. The results indicate that CLA significantly reduces inflammation compared to corn oil fed arthritic mice and that the anti-inflammatory effects of 0.5% c9t11-CLA are equal to a celecoxib dose in between 5 and 50mg/kg bw, with 5mg/kg being the commonly prescribed daily dosage for humans. The results of this experiment indicate that humans may receive anti-inflammatory benefits from the consumption of CLA in food products such as dairy fat without the excess costs and burdens of prescription drugs. A research presentation entitled, "How might dietary fat intake affect your use of NSAIDs?" was presented by Jake Olson at the August 30th Molecular and Environmental Toxicology Center seminar at UW-Madison. 3. Compare the inflammatory metabolome of mice fed c9, t11- CLA verses COX-2 inhibitor. Plasma samples collected from the previous experiment involving dietary treatment with three doses of celecoxib and c9t11-CLA are currently being analyzed by Jake Olson and co-PI Fariba Assadi-Porter to further evaluate whether these dietary treatments alter systemic metabolites, such as restoring the hypoaminoacidemia seen in the previous examination of plasma metabolites in arthritic mice. Since celecoxib has been shown to cause comorbidities in humans and c9t11-CLA has not, an evaluation of metabolites may lead to the discovery of previously unknown potential side effects or mechanisms of action for both celecoxib and c9t11-CLA. 4. Determine the interaction of CLA and COX inhibitors in preventing inflammation. Due to the finding that a dietary amount of 0.5% c9t11-CLA was more effective at reducing inflammation than the relevant daily dose (5mg/kg) of celecoxib, we are further pursuing the minimum effective dose of c9t11-CLA in reducing inflammation such that a lower dose would translate to less CLA consumption by humans, while maintaining a meaningful reduction of inflammation and therefore, a more practical means of controlling chronic inflammation than a concomitant treatment with both c9t11-CLA and celecoxib. A dose response experiment feeding c9t11-CLA at and below 0.5% of total diet has been performed and the results of this experiment indicate that 0.125% c9t11-CLA is as effective as 0.5% after 42 days of treatment. This dosage translates to a daily human dose of 3.025g and may not be the absolute minimum effective dose. Therefore, our results indicate that human consumption of a dietary source (e.g. dairy fat) rich in c9t11-CLA may be an effective and highly practical means of controlling chronic inflammation in humans.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Huebner SM, Olson JM, Campbell JP, Bishop JW, Crump PM, Cook ME. (2014) Dietary trans-10,cis-12 CLA Reduces Murine Collagen-Induced Arthritis in a Dose-Dependent Manner. J Nutr. doi: 10.3945/jn.113.182550
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Progress 01/01/13 to 09/30/13
Outputs
Target Audience: Health professionals, scientists, animal nutritionists, livestock and dairy farmers, and the general public are the targeted audiences for this work. In the current reporting period, information gathered as a result of this project have been shared with all audiences through presentations, publications, and services such as CLA analyses of animal products. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Experiments have led to the development of new hypotheses and therefore new research opportunities. How have the results been disseminated to communities of interest? Results have been disseminated to the scientific community through formal presentations, publications, and research proposals. Specifically, a research presentation entitled, “In search of the anti-inflammatory components in dairy fat” was given by Jake Olson at the October 24th Molecular and Environmental Toxicology Center seminar at UW-Madison. A manuscript entitled, “Dietary trans-10,cis-12 CLA Reduces Murine Collagen-Induced Arthritis in a Dose-Dependent Manner” was accepted for publication by the Journal of Nutrition and will be in print February of 2014. Our lab works closely with the agriculture industry through farmers and dairy producers, such that we provide analyses of products for CLA isomer content and further consult with interested parties to develop feeding strategies for CLA enrichment. What do you plan to do during the next reporting period to accomplish the goals? We have established an essential metabolomics analysis software in our laboratory. Remaining animal experiments will take place throughout the next reporting period and a final analysis of the data will be conducted in 2014. Furthermore, a comprehensive metabolomic analysis will be performed as per objective 3.
Impacts
What was accomplished under these goals?
Since our last progress report, several accomplishments have been achieved. A research presentation entitled, “In search of the anti-inflammatory components in dairy fat” was given by Jake Olson at the October 24th Molecular and Environmental Toxicology Center seminar at UW-Madison. A manuscript entitled, “Dietary trans-10,cis-12 CLA Reduces Murine Collagen-Induced Arthritis in a Dose-Dependent Manner” was published by Journal of Nutrition. Included in this dose response experiment is the calculation that anti-inflammatory levels of t10c12-CLA are not attainable from whole food sources in a practical diet without supplementation. Establishing a t10c12-CLA dietary anti-inflammatory threshold allows us to set and assess relevant or irrelevant dosages in future experiments. Furthermore, we have shown plasma cytokines (e.g. IL-1β and TNFα) to be unreliable biomarkers of chronic inflammation and demonstrated paw cytokines to be valid indicators of the inflammatory state in arthritic mice. Metabolomic analysis has been completed under objective 1 and a manuscript is currently in preparation.
Publications
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2013
Citation:
Huebner SM, Olson JM, Campbell JP, et al. (2013) Dietary trans-10,cis-12 CLA Reduces Murine Collagen-Induced Arthritis in a Dose-Dependent Manner. J Nutr. doi: 10.3945/jn.113.182550
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: Since our last progress report, several outputs have been achieved. A research presentation entitled, "Metabolic analyses of specific isomers of conjugated linoleic acid in the collagen induced arthritis model" was given by Jake Olson at the November 15th Molecular and Environmental Toxicology Center seminar at UW-Madison. A symposium entitled "Conjugated Linoleic Acid (CLA): A Naturally Occurring Anti-Obesity, Anti-Inflammatory Fatty Acid" was developed in honor of Dr. Michael W. Pariza, who first isolated CLA, at the American Chemical Society's August national meeting in Philadelphia, PA; where he was awarded the 2011 Kenneth A. Spencer Award. At this event, Prof. Mark Cook gave a presentation entitled, "Dietary CLA reduces clinical signs of acute and chronic inflammatory disease." A poster presentation entitled, "Development of a chronic inflammation metabolome and cytokine profile to screen potential pro and anti-inflammatory compounds" was presented by Jake Olson on May 23rd at a WARF Discovery Challenge event at UW-Madison. Recently, we analyzed several livestock products to determine CLA content for SunRay Dairy, LLP, a Wisconsin dair, marketing CLA as a health-promoting natural product. This is one example of many in which dairy producers now market CLA for its health benefits. PARTICIPANTS: Professor Mark E. Cook (Animal Sciences Dept., UW) serves as the principle investigator on the project. CO-PIs Drs. F.M. Assadi-Porter (National NMR Center, Biochemistry, UW) and D.E. Butz (NMR Center and Animal Sciences, UW) are primarily responsible for the NMR analyses. Dr. Changhua Li, visiting professor has been responsible for developing new methods of sample preparation for NMR analyses. Dr. S Huebner, Jeffery Bishop, and Dr. J Sand have been supportive of training a new graduate student, Jake Olson, in animal studies and target analyses. Based on our previous results, we are discussing several applications of both CLA and metabolomic analyses in inflammatory diseases, such as rheumatoid arthritis, with Daniel Muller, MD, PhD, a clinical rheumatologist. Opportunities are consistently available for undergraduates to participate in this research project. An undergraduate student, Alexander Haas, has been hired as an undergraduate student hourly employee to assist Jake Olson in completing the objectives of this project. The undergraduate student will have opportunities to apply for small supplementary grants to further support this project, as well as to develop personally as a researcher. TARGET AUDIENCES: Health professionals, scientists, animal nutritionists, livestock and dairy farmers, and the general public are the targeted audiences for this work in the long-term. In the current reporting period, information gathered as a result of this project has been shared with all audiences through presentations, publications, and services such as CLA analyses of animal products. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Based on the complexity of metabolomic data analyses, we are currently developing techniques and strategies to interpret large amounts of metabolomic data. Dr. Changhua Li, a visiting professor, continues to help develop methods for sample preparation using nuclear magnetic resonance spectroscopy (NMR). We are also currently collaborating with Associate Professor Fariba Assadi-Porter to develop appropriate statistical models for analyses of metabolomic data. Animal experiments to advance our objectives are in progress at this time. Results from our previous research have led to several outcomes/impacts. Prof. Mark Cook was invited to present at a symposium at the American Chemical Society's national meeting in Philadelphia, where he gave a presentation entitled, "Dietary CLA reduces clinical signs of acute and chronic inflammatory disease." Furthermore, at this event Dr. Michael W. Pariza, a past collaborator, was awarded the Kenneth A. Spencer award for his research on CLA. Based on our previous results, we are discussing several novel applications of both CLA and metabolomic analyses in inflammatory diseases, such as rheumatoid arthritis, with Daniel Muller, MD, PhD, a clinical rheumatologist at UW-Madison.
Publications
- Cherian, G and Poureslami, R. (2012). Fats and Fatty Acids in Poultry Nutrition and Health. Paul and Company Pub Consortium.
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: The primary goals of this research project are to develop a nuclear magnetic resonance metabolome of plasma using an arthritic mouse model that would be useful in diagnosis chronic inflammation. Additional objectives were to determine the effectiveness cis 9, trans 11 conjugated linoleic acid (c9t11-CLA), a naturally occurring animal fat, as an effective treatment of chronic inflammation without causing adverse effects on secondary nutrient metabolism (as determined by the NMR metabolome). While only one year has been completed on this specific project, several outputs have been achieved. First, publication of scientific papers in the past on c9t11-CLA's anti-inflammatory effects have resulted in the recognition that the consumption of ruminant fat, found in milk or beef, may have anti-inflammatory health benefits. This has been evident in scientific publications linking the consumption of ruminant fats to decreased inflammatory diseases such as heart disease. Increasingly, dietary practices for cattle that increase the levels of c9t11-CLA or its precursors in ruminant fat are being promoted as having health benefits. In addition, analysis of the NMR metabolome with regards to CLA consumption provides evidence of CLA's safety. This year, a webinar was sponsored by International Food Technologists on the safety of CLA for animal and human consumption. Interest in this topic also resulted in an invited presentation at the American Oil Chemist Society meeting in the session on Lipids and Inflammation. Implications and limitations of the use of CLA in poultry were also presented at a scientific meeting and by invitation to the Mid-Atlantic Nutrition Conference. Also, while not directly funded by this project, a patent for the use of the NMR inflammatory metabolome in the detection of chronic inflammation was awarded by the U.S. Patent Office. PARTICIPANTS: Professor Mark E. Cook (Animal Sciences Department, UW) serves as the principal investigator on the project. Co-PIs Drs. F.M. Assadi-Porter (National NMR Center, Biochemistry, UW) and D.E. Butz (NMR Center and Animal Sciences, UW) are primarily responsible for the NMR analyses. Dr. Changhua Li, visiting professor (in Cook's lab) has been responsible for developing new methods of sample preparation for NMR analysis. Dr. M. Tonelli (NMR Center) has provided NMR instrument support. Professor H. Eghbalnia, Department of Molecular and Cellular Physiology, University of Cincinnati has provided advice on data computation. Dr. S. Huebner and Jeffery Bishop have been supportive in training a new graduate student, Jake Olson (Cook's lab) in animal studies and data analysis. TARGET AUDIENCES: Health professionals, scientists, animal nutritionist, and the general public are the targeted audiences for this work in the long-term. In the current reporting period, information gathered as a result of this specific project have been shared with scientists through presentations and publications. A presentation was delivered at a nutritional conference in Maryland that specifically targeted animal nutritionists. A press release in UW's Grow Magazine provided a short article on the anti-inflammatory effects of CLA. The webinar hosted by IFT was viewed, on the first day, by over 50 individuals or groups with interest in food science or food product development. PROJECT MODIFICATIONS: No major changes were made to the objectives. In the first year of this project, we were unable to identify a suitable graduate student to start the project. Activities for the first year of the project was managed by another graduate student and a visiting professor. A new student, Jake Olson were identified in the fall of 2011.
Impacts
Key outcomes to date that have resulted from this research are that chronic inflammatory disease such as arthritis causes disruption in the NMR metabolome. Significant changes involve the plasma pool of amino acids. Chronic arthritis causes a hypoaminoacidemia. The aberrant amino acid metabolome was restored to normalcy when conjugated linoleic acid was included in the diet. There are several impacts from these findings. First, secondary diseases (i.e., cardiovascular disease) that have been associated with chronic inflammation may be caused by alterations in nutrient metabolism. Correcting the changes in nutrient metabolism by using a naturally occurring fatty acid such as CLA could potentially improve the outcome of the people or animals that suffer from chronic inflammation. In a recent study out of Harvard University, the level of c9t11-CLA in humans was inversely related to the risk of myocardial infarction; a finding that may be linked to reduced inflammation. Second, the development of a NMR metabolome that is diagnostic of inflammation will allow the determination of subclinical forms of inflammation that may affect human of animal health. Inflammation is considered a silent killer (Times Magazine). The problem has been the identification of individuals with subclinical inflammation. Select biomarkers have been used as indices of subclinical inflammation (i.e., c-reactive protein or CRP), however these biomarkers have often resulted in misleading diagnoses and some diseases that elevate CRP may not actually be forms of chronic inflammation (for example, obesity). Powerful tools for measuring chronic inflammation will impact decisions in the treatment of patients. Third, the finding that CLA in ruminant fat may restore the inflammatory metabolome to normalcy suggests that products of ruminant origin may actually be beneficial to human health. The impact of these finding could alter long-term dietary recommendation for humans. Another impact could be a the farm level where ruminant nutritionist design feeding programs that assure CLA levels in ruminant products that could improve health outcomes. By securing a patent on the use of NMR metabolomics as a measure of inflammation, researchers can make sure the use of the technology, at least for a limited time, is used in a manner that agrees with scientific findings.
Publications
- Cook, M.E., and Sand J.M. (2011).What poultry nutritionists and marketers need to know about conjugated linoleic acid: a potent anti-inflammatory of ruminant origin.Proc. 9th Ann. Mid-Atlantic Nutr. Conf., pp. 169-177.
- Leone, V.A., Haughey, D., Bobeck, E.A., Cook, M.E., and Assadi-Porter, F.M. (2011. Evidence that maternal conjugated linoleic acid alters secondary metabolites in plasma of late stage embryos that may lead to increased embryonic mortality. J. Anim. Sci. 89. E-suppl:357.
- Cook, M.E., and Huebner, S.M. (2011). Conjugated linoleic acid's anti-inflammatory action in an animal model of rheumatoid arthritis. Am. Oil Chem. Soc. Ann. Meeting. E-abstract http://www.aocs.org/archives/am2011/abstracts.cfminterest=Health%20a nd%20Nutrition.
- Assadi-Porter, F.M., Cook, M.E., Eghbalnia, H.R., Tonelli, M. Porter, W.P., Butz, D.E. (2011). Noninvasive measurement and identification of biomarkers in disease state. U.S. Patent 8026049.
- Pariza, M.W., Park, Y., and Cook, M.E. (2011). Conjugated linoleic acid from discovery to GRAS, a 20 year journey.Intern. Food Technologist webnair. http://www.ift.org/knowledge-center/learn-online/on-demand-webcasts/f ood-health-and-nutrition/conjugated-linoleic-acid.aspx
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: This project began October 1, 2010. This project is related to completed project WIS01073. No outputs have been created since the three months of this project's start. PARTICIPANTS: Professor Mark E. Cook (Animal Sciences Department, University of Wisconsin) will serve as the principal investigator on the project. Dr Fariba Assadi-Porter (Biochemistry, University of Wisconsin)and Dr. Daniel E. Butz (Zoology and Animal Sciences Department, University of Wisconsin) will serve as co-investigators on the project and will assist in metabolomic analyses. Professor Hamid Eghbalnia, Department of Molecular and Cellular Physiology, University of Cincinnati will assist in data computation. Dr. Shane Huebner (Department of Nutritional Sciences, University of Wisconsin) will provide training in techniques involving the animal model. Jeffery Bishop, an undergraduate in Biochemistry, University of Wisconsin, will assist in the animal experimentation. TARGET AUDIENCES: A meeting with a science communicator, Nicole Miller (Office of External Affairs, College of Agricultural and Life Sciences, University of Wisconsin), has occurred. The plan is to draft an article on the findings from WIS01073 and anticipated results from this project for the general public. A meeting is scheduled January 7, 2011 with a medical doctor in Rheumatology to explore grant funding to conduct clinical trials. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
During the three months period of the project, graduate students have rotated through the laboratory. These students gained hands on experience working with different aspects of the project.
Publications
- No publications reported this period
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