Progress 01/01/14 to 12/31/16
Outputs
Target Audience:The central target audience of this project are producers that manage dairy cows. Although the research will benefit dairy producers nationally, a more immediate impact has been made regionally within Somerset County, PA. Academic, government, and industry professionals also serve as a target audience. At West Virginia University (WVU), the target audience has included undergraduate and graduate students seeking a career in the agricultural sciences, and faculty interested in employing a mass spectrometry-based metabolomics approach for their own research goals. At WVU (for 2016), training exercises included blood marker colorimetric and mass spectrometry analysis, tissue lipid extraction, immunoblotting, and continued data organization, statistical analysis, and presentation. Additionally, mass spectrometry-based methodologies that were developed with this project were utilized by other biomedical research programs at WVU. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A total of two Ph.D. students, two M.S. students, and two undergraduate researchers were mentored during Year 3. These students were trained to analyze serum and liver ceramide levels, liver lipid content, blood fatty acid, glucose, cholesterol, ketones, and LDL concentrations. Students were also trained to statistically analyze their data and prepare manuscripts for peer review. How have the results been disseminated to communities of interest?The community of interest are individuals within the dairy industry including but not limited to dairy farmers, veterinarians, nutrition companies, academic researchers, county extension agents, and government agricultural researchers. A series of scientific abstracts were accepted for presentation at the 2016 ADSA-ASAS Joint Annual Meeting, in addition to local and regional conferences and student competitions. Two manuscripts were submitted for peer review. We have also shared our results with the herd manager, veterinarian, and nutritionist at DoVan Farms. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The transition from gestation to lactation represents a critical period in the life cycle of the dairy cow. During this timeframe, dairy cows will experience a physiological state of negative energy balance characterized by excessive fatty acid concentrations in circulation (caused by adipose tissue mobilization) and hepatic ketogenesis. In turn, dairy cows can develop metabolic diseases such as fatty liver and ketosis during early lactation. The occurrence of metabolic diseases is greater in overweight cows, a result attributed to greater fatty acid levels in blood and reduced insulin sensitivity. Identifying the cause of insulin resistance in overweight dairy cows and developing new strategies to control insulin action to lower circulating fatty acid levels may be a means to improve animal health, reduce costs, and improve dairy farm profitability. In addition to finalizing a dissertation, we also submitted two manuscript for peer review. We also had the opportunity to analyze new data related to our second in vivo study (i.e. feed restriction study). Specifically, we tested the hypothesis that ceramide (ceramideaccumulates in dairy cows experiencing lipolysis and insulin resistance. To recap, nine dairy cows were utilized in a replicated 3×3 Latin square design. Cows were ad libitum fed, nutrient-restricted (NR), or NR with nicotinic acid (NA; 5 mg of NA/h per kg BW; delivered i.v.) for 34 h. When provided access, cows were ad libitum fed a mixed ration of grass hay and ground corn to meet requirements. Intake for NR cows was limited to vitamins and minerals. Nicotinic acid was administered to suppress lipolysis. Saline was infused in cows not provided NA. At h 32 and 33 of treatment, a liver biopsy and insulin tolerance test were performed, respectively. Samples were analyzed using colorimetry, immunoassay, and mass spectrometry. Nutrient restriction increased serum fatty acids and ceramide levels, and impaired insulin sensitivity; however, NA infusion was unable to prevent these responses. We also show that NR increases hepatic ceramide accumulation, a response that was positively associated with serum ceramide supply. Our data demonstrate that circulating and hepatic 24:0-ceramide are inversely associated with systemic insulin tolerance, an effect not observed for the 16:0 moiety. In conclusion, our results suggest that ceramide accrual represents a metabolic adaptation to nutrient restriction and impaired insulin action in dairy cows.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Rico, J. E., F. Seck, M. V. Pinti, and J. W. McFadden. "1075 Increasing fatty acid oxidation improves insulin sensitivity in primary differentiated bovine adipocytes." Journal of Animal Science 94, no. supplement5 (2016): 515-515.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Davis, A. N., J. L. Clegg, and J. W. McFadden. "1335 Intravenous nicotinic acid suppresses adipose tissue lipolysis in Holstein dairy cows." Journal of Animal Science 94, no. supplement5 (2016): 644-644.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Samii, S. Saed, J. E. Rico, and J. W. McFadden. "1513 Characterization of peripartum liver and skeletal muscle ceramide concentrations in lean and overweight Holstein dairy cows." Journal of Animal Science 94, no. supplement5 (2016): 734-735.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2016
Citation:
SPHINGOLIPID METABOLISM IN DAIRY COWS AND ITS RELATIONSHIP TO CIRCULATING LIPIDS AND INSULIN RESISTANCE
- Type:
Journal Articles
Status:
Submitted
Year Published:
2016
Citation:
Temporal Changes in Sphingolipids and Insulin Sensitivity during the Transition from Gestation to Lactation
- Type:
Journal Articles
Status:
Under Review
Year Published:
2016
Citation:
Nutrient restriction increases circulating and hepatic ceramide in dairy cows displaying impaired insulin tolerance.
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Progress 01/01/15 to 12/31/15
Outputs
Target Audience:The central target audience of this project are producers that manage dairy cows. Although the research will benefit dairy producers nationally, a more immediate impact has been made regionally within Somerset County, PA. Academic, government, and industry professionals also serve as a target audience. At West Virginia University (WVU), the target audience has included undergraduate and graduate students seeking a career in the agricultural sciences, and faculty interested in employing a mass spectrometry-based metabolomics approach for their own research goals. Further efforts included training undergraduate and graduate students to manage and execute an in vitro scientific study. Training exercises included tissue collection, cell growth, treating cells, harvesting and storing cells, cell based assays, and data interpretation. Training exercises related to in vivo experimentation included animal care, sample (blood, liver, muscle, and adipose) collection, feed formulation, animal health diagnostics, milking technique, and biological sample harvesting and storage. Chemistry training exercises included blood marker colorimetric and mass spectrometry analysis, and tissue lipid extraction. Additional training in the areas of data organization and statistical analysis were provided. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A total of two Ph.D. students, two M.S. students, and three undergraduate researchers were mentored during Year 2. These students were trained by the PD to perform liver biopsies, collect blood and prepare samples for storage, record adiposity, collect feed and milk samples, record observations, insert jugular catheters, perform insulin tolerance testing, perform colorimetric (glucose, NEFA, ketones) and immonoassays (insulin, interleukins), execute mass spectrometry protocols (non-targeted and targeted using GC/MS and LC/MS technologies), manage metabolomics data sets, culture primary bovine adipocytes, and statistically analyze large data sets using multiple procedures. How have the results been disseminated to communities of interest?The community of interest are individuals within the dairy industry including but not limited to dairy farmers, veterinarians, nutrition companies, academic researchers, county extension agents, and government agricultural researchers. A series of scientific abstracts were accepted for poster and oral presentation at the 2015 ADSA-ASAS Joint Annual Meetings, in addition to local and regional conferences and student competitions. We have also shared our results with the herd manager, veterinarian, and nutritionist at DoVan Farms. What do you plan to do during the next reporting period to accomplish the goals?We need to statistically analyze and prepare in vivo and in vitro data for a manuscript submission and peer review.
Impacts
What was accomplished under these goals?
The transition from gestation to lactation represents a critical period in the life cycle of the dairy cow. During this timeframe, dairy cows will experience a physiological state of negative energy balance characterized by excessive fatty acid concentrations in circulation (caused by adipose tissue mobilization) and hepatic ketogenesis. In turn, dairy cows can develop metabolic diseases such as fatty liver and ketosis during early lactation. The occurrence of metabolic diseases is greater in overweight cows, a result attributed to greater fatty acid levels in blood and reduced insulin sensitivity. Identifying the cause of insulin resistance in overweight dairy cows and developing new strategies to control insulin action to lower circulating fatty acid levels may be a means to improve animal health, reduce costs, and improve dairy farm profitability. We analyzed data collected from our 2014 in vivo study that evaluated the relationship between the bovine lipidome and systemic insulin action in lean and overweight dairy cows transitioning from gestation to lactation. Our results demonstrated the following: first, insulin resistance develops postpartum concurrently with increased lipolysis and hepatic lipid accumulation; second, ceramides and glycosylated ceramides accumulate during the transition from gestation to lactation and are further elevated in overweight cows; third, ceramide accrual is associated with lipolysis and liver lipid accumulation, and C16:0 and C24:0-ceramide were inversely associated with systemic insulin sensitivity postpartum; fourth, plasma sphingomyelin, a potential source of ceramides reached a nadir at parturition and were closely associated with feed intake; fifth, select sphingomyelins were lower in the plasma of overweight cows during the peripartal period sixth, insulin resistance indices do not reflect changes in systemic insulin or glucose tolerance. Our results demonstrate that dynamic changes occur in peripartal sphingolipids that are influenced by adiposity, and are associated with the onset of peripartal insulin resistance. These observations are in agreement with a putative potential role for sphingolipids in facilitating the physiological adaptations of peripartum. The progression of insulin resistance in dairy cows represents a maternal adaptation to support milk production during heightened energy demand; however, excessive adipose tissue lipolysis can develop. In diabetic monogastrics, the mechanisms that mediate insulin resistance involve the sphingolipid ceramide. Considering that we previously demonstrated that ceramides are elevated during the peripartum, and intravenous lipid infusion increases circulating and hepatic ceramide, we explored the relationship between nutrient restriction and ceramide supply. We also evaluated the effects of nicotinic acid, a known suppressant of lipolysis, on ceramide supply during nutrient restriction. We tested the hypothesis that ceramide accumulates in dairy cows experiencing lipolysis and insulin resistance. Nine dairy cows were utilized in a replicated 3×3 Latin square design. Cows were ad libitum fed, nutrient-restricted (NR), or NR with nicotinic acid (NA; 5 mg of NA/h per kg BW; delivered i.v.) for 34 h. When provided access, cows were ad libitum fed a mixed ration of grass hay and ground corn to meet requirements. Intake for NR cows was limited to vitamins and minerals. Nicotinic acid was administered to suppress lipolysis. Saline was infused in cows not provided NA. At h 32 and 33 of treatment, a liver biopsy and insulin tolerance test were performed, respectively. Samples were analyzed using colorimetry, immunoassay, and mass spectrometry. Assay of blood (basal or following an insulin challenge) and liver variables (health markers and ceramides) is still required. Dairy cows develop insulin resistance during the transition from gestation to lactation. Because insulin is an anti-lipolytic hormone, insulin resistance promotes adipose tissue lipolysis. Increasing fatty acid oxidation (FAox) is a means to improve insulin sensitivity in monogastrics. Therefore, our second objective was to evaluate the effects of a pharmacological stimulator of FAox on insulin sensitivity in bovine adipocytes. To test our objective, we utilized subcutaneous adipose tissue collected from Angus steers. Stromal-vascular cells were grown from explants in DMEM/F12 growth medium containing 17.5 mM glucose and 10% fetal bovine serum (FBS). Cells were harvested by trypsinization and replated. Once confluent, cells were differentiated using DMEM/F12 medium containing 17.5 mM glucose, 5 mM acetate, 1 mM octanoate, 5% FBS, 1.72 mM insulin, 0.25 mM dexamethasone, 0.5 mM isobutylmethylxanthine, and 2 mM rosiglitazone. Following d 8 of differentiation, cells were cultured in DMEM/F12 treatment medium containing 5 mM glucose, 1 mM acetate, 1% FBS, and 1.2 nM insulin. Differentiated adipocytes were treated with C75 (stimulator of carnitine palmitoyltranserase 1 and FAox; 0 to 100 µM) or palmitic acid (C16:0; 0 to 800 µM) complexed with bovine serum albumin for 3 to 18 h. Control cells did not receive C75 or C16:0 treatment. A minimum of two independent experiments with three replicates per experiment were performed. The statistical model included the fxed effects of treatment, experiment, and their interaction. Replicate within experiment was the random effect. Triacylglycerol (TAG) accumulation and cell viability were determined using colorimetric and fuorescence assays, respectively. Measurement of FAox and insulin sensitivity were measured using radiolabeled 2-deoxyglucose (2DOG) and C16:0. Relative to undifferentiated adipocytes, TAG accumulation was 736% greater in differentiated adipocytes. Following differentiation, treatment with C16:0 or C75 for 18 h did not impair cell viability. Interestingly, 100 mM C75 improved cell viability by 40%, relative to control. Treatment of adipocytes with 100 mM C75 for 3 h increased FAox, as demonstrated by a 122% increase in the recovery of radiolabeled acid-soluble products as well as a 30% increase in radiolabeled CO2. Although C16:0 did not modify insulin 2DOG uptake following an 18 h treatment, treating adipocytes with 100 mM C75 for 18 h increased 2DOG uptake by 141%, relative to control. We conclude that the stimulation of FAox enhances insulin-stimulated glucose uptake in primary differentiated bovine adipocytes.
Publications
|
Progress 01/01/14 to 12/31/14
Outputs
Target Audience: The central target audience of this project are producers that manage dairy cows. Although the research will benefit dairy producers nationally, a more immediate impact has been made regionally within Somerset County, PA. Academic, government, and industry professionals also serve as a target audience. At West Virginia University (WVU), the target audience has included undergraduate and graduate students seeking a career in the agricultural sciences, and faculty interested in employing a mass spectrometry-based metabolomics approach for their own research goals. For four-months (May to August of 2014), the WVU project team was located at DoVan Farms (Berlin, PA), a 700-cow Holstein dairy farm owned and operated by the VanGilder family. This farm is a WVU Agricultural Research and Education Partner and is the external site of the described in vivo research (Aim 1 sample collection only). During the project duration, students and the project leader delivered educational workshops to the VanGilder family. Workshops included transition cow health and potential implications for the described research. Because Aim 1 sample collected occurred at DoVan Farms and the project team lived in Berlin, PA for the duration of the study, the graduate students had the opportunity to learn modern dairy production and management practices from the VanGilder family. Further efforts included training undergraduate and graduate students to manage and execute a scientific study located on a commercial dairy farm. Training exercises included animal care, sample (blood, liver, muscle, and adipose) collection, feed formulation, animal health diagnostics, milking technique, sample harvesting and storage, and data organization. At WVU, training exercises included blood marker colorimetric and mass spectrometry analysis, tissue lipid extraction, mRNA gene expression analysis, continued data organization, statistical analysis, and presentation. Additionally, mass spectrometry-based methodologies that were developed with this project were used as a training exercise in a biochemistry wet lab offered to undergraduates at WVU. Changes/Problems: One challenge that we faced was our inability to collect feed intake data past d 6 postpartum. Because we chose to perform our research on a commercial dairy farm, we installed Calan gates in a special pen on the farm for feed intake monitoring; however, due to space constraints we had to move these animals to a free-stall system after d 6 in milk. Although we faced limitation, we were still able to document greater reductions in appetite in overweight cows at parturition. For future work, this problem has been addressed. Starting in 2015, DoVan Farms has now provided WVU with additional barn space for transition cow research and inidivudal pens have been installed. We can now house up to 20 cows within inidvidual pens at any one time and for any duration of time. Time and space constraints is no longer an issue. No other problems existed. What opportunities for training and professional development has the project provided? A total of two Ph.D. students, two M.S. students, and three undergraduate researchers were mentored during Year 1. These students were trained by the PD to perform liver, adipose, and muscle biopsies, collect blood and prepare samples for storage, record adiposity, collect feed and milk samples, record observations, insert jugular catheters, perform insulin and glucose tolerance testing, perform colorimetric (glucose, NEFA, ketones) and immonoassays (insulin, interleukins), execute mass spectrometry protocols (non-targeted and targeted using GC/MS and LC/MS technologies), manage metabolomics data sets, culture primary bovine adipocytes, initiate gene expression analysis protocols (i.e. RNA isolation, cDNA synthesis, and PCR), and statistically analyze large data sets using multiple procedures. All students have submitted abstracts for national presentation, and all have participated in local poster/oral talk competitions. As a result, multiple students received scholastic awards. Rachel Cokeley (Outstanding Senior Award), Eduardo Rico (two-time 2nd place poster competition award), Sina Saed Samii (1st place poster competition award), Amanda Davis (Ruby Distinguished Doctoral Fellow). How have the results been disseminated to communities of interest? The community of interest are individuals within the dairy industry including but not limited to dairy farmers, veterinarians, nutrition companies, academic researchers, county extension agents, and government agricultural researchers.(1) To reach the local community, our research team hosted a table at the Somerset County Fair in Somerset, PA for a 1-week duration. We presented the rationale for our research, current data, and potential impact. We also educated the local community about the agricultural research and education partnership between West Virginia University and DoVan Farms (a commercial dairy farm) located in Somerset County, PA. (2) To education dairy industry professionals about the field of metabolomics and the potential impact these technologies may have on the future of dairy production, a popular press article in Progressive Dairyman was published. (3) A series of scientific abstracts were accepted for poster and oral presentation at the 2014 and 2015 ADSA-ASAS Joint Annual Meetings, in addition to local and regional conferences and student competitions. (4) We have shared our results with the herd manager, veterinarian, and nutritionist at DoVan Farms. (5) One manuscript is currently being reviewed by the Journal of Dairy Science, while a minimum of three other manuscripts are in preparation for manuscript submission for peer-review. What do you plan to do during the next reporting period to accomplish the goals? To complete the project goal for Objective 1 we will: 1. Measure plasma and tissue free fatty acid concentrations. 2. Quantify liver lipid accumulation. 3. Evaluate changes in gene expression and protein abundance of tissues with a focus on ceramide and fatty acid metabolism, inflammatory signaling, and insulin action. 4. Perform non-targeted GC/MS analyses on plasma and liver tissues. 5. Explore new methods to quantify different metabolite classes (e.g. isoprostanes or steroids). 6. Statistically analyze our large data sets to draw associations between the bovine metabolome, adiposity, and insulin resistance in periparturient dairy cows. 7. Finally, submit our in vivo work for peer-review publication. To complete the project goal for Objective 2 we will: 1. Validate our in vitro primary bovine adipocyte insulin resistance model. 2. Utilize pharmacological modifiers of fatty acid metabolism to increase fatty acid oxidation and improve insulin sensitivity in insulin resistance bovine adipocytes. 3. Characterize the adipocyte metabolome (with a focus on ceramide and fatty acid levels) in insulin sensitive and resistance cells in the absence or presence of pharmacological modifiers that can augment fatty acid oxidation. 4. Analyses will include 2-deoxyglucose measurement of insulin sensitivity, measurement of lipolysis, gene expression and immunoblotting of genes related to ceramide metabolism and insulin action, respectively, as well as mass spectrometry analyses of cellular metabolite pools. 5. Finally, submit our in vitro work for peer-review publication. Additional work: 1. We are also continuing our collaborative work with Dr. Yves Boisclair. Because we demonstrated an increase in circulating ceramides following i.v. infusion with a soybean triglyceride emulsion (also increasing NEFA availability), we are measuring the liver expression of genes related to ceramide metabolism in samples collected from these cows to determine the impact of excess NEFA availability on hepatic gene expression profiles related to ceramide synthesis. We are also characterizing the free fatty acid profiles of plasma collected from these animals.
Impacts
What was accomplished under these goals?
The transition from gestation to lactation represents a critical period in the life cycle of the dairy cow. During this timeframe, dairy cows will experience a physiological state of negative energy balance characterized by excessive fatty acid concentrations in circulation (caused by adipose tissue mobilization) and hepatic ketogenesis. In turn, dairy cows can develop metabolic diseases such as fatty liver and ketosis during early lactation. The occurrence of metabolic diseases is greater in overweight cows, a result attributed to greater fatty acid levels in blood and reduced insulin sensitivity. Identifying the cause of insulin resistance in overweight dairy cows and developing new strategies to control insulin action to lower circulating fatty acid levels may be a means to improve animal health, reduce costs, and improve dairy farm profitability. 1. For Objective 1: First (January to May of 2014), we utilized an existing sample set limited to plasma samples collected from lean and overweight periparturient cows to identify novel biomarkers associated with hyperlipidemia and insulin resistance. Samples were collected at -30, -15, -7, and 4 d relative to calving. Utilizing LC/MS in collaboration with Dr. Norman Haughey at Johns Hopkins University and GC/MS in the lab of the PD at West Virginia University, we have profiled (detected) 32 ceramides (including monohexosylceramides and lactosylceramides), 18 sphingomyelins, 4 long-chain fatty acylcarnitines, 32 free fatty acids, and 40 metabolites using our GC/MS-based non-targeted metabolite profiling approach. A major finding was that ceramide levels in plasma were higher in overweight transition cows. Furthermore, the level of ceramide increased during the transition in all cows, suggesting a homeorhetic response to a change in physiological state. Also of interest, we observed a delayed increase in glycosylated ceramide subspecies in all animals with elevated ceramide concentrations in plasma. Furthermore, we have discovered a positive correlation between NEFA availability and ceramide accumulation in plasma (e.g. C24:0-ceramide). We also demonstrated a negative correlation between ceramide accumulation in plasma and insulin sensitivity (e.g. C24:0-ceramide). Similar observations were observed for saturated fatty acids (e.g. palmitic acid) and long-chain fatty acylcarnitines (e.g. C16:0-acylcarnitine). In contrast, we discovered a negative correlation between NEFA availability and sphingomyelin accumulation in plasma. As well as a positive correlation between sphingomyelin accumulation in plasma and insulin sensitivity. Lastly, plasma ceramides were negative correlated with sphingomyelin levels. These key findings support our hypothesis that fatty acid oxidation is impaired in overweight transition cows which supports the de novo synthesis and accumulation of the sphingolipid ceramide. It should also be noted that our sphingolipid profiles in overweight, insulin resistant cows mimicked those observed in overweight, type 2 diabetic humans. To further explore the relationship between NEFA availability, ceramide accumulation, and insulin action in cows we A) executed Objective 1 with a focus on ceramide metabolism and insulin signaling, and B) collaboratively assessed the relationship between NEFA supply and ceramide accumulation in cows experiencing acute increases in plama NEFA. A. Sample collection: From May to September of 2014, we completed the in vivo sample collection for Objective 1. At DoVan Farms, WVU Agricultural Research and Education Partner (a 700 Holstein cow dairy farm in Berlin, PA), we enrolled 14 multiparous, Holstein dairy cows experiencing overnutrition or nutrient restriction (animals displaying an overweight or lean phenotype, respectively). During the transition from gestation to lactation (-28 to 28 d relative to calving), blood and milk were collected routinely. Additionally, tissue (adipose, muscle, and liver) were biopsied at -21, -7, and 4 d relative to calving. An insulin tolerance test was performed at -20, -6, and 5 d relative to calving. A glucose tolerance test was performed at -19, -5, and 6 d relative to calving. Individual intakes were monitored, and body weight and adiposity were measured weekly. Feed samples were also collected and composites formed bi-monthly. Milk production was also recorded. Major data acquired: We observed greater concentrations of glucose, insulin, ketones (i.e. BHBA), and fatty acids in cows experiencing overnutrition. Overweight cows experienced greater appetite suppression at parturition, and greater reductions in glucose-stimulated NEFA disposal. Insulin tolerance data also demonstrated greater reductions in insulin-stimulated glucose disposal postpartum, as compared to prepartum testing. Evidence for impaired hepatic gluconeogenesis was also observed in overweight cows. From a metabolomics perspective, we observed dramatic temporal changes in plasma sphingolipid levels in all cows transitioning from gestation to lactation. For example, we observed a sustained increase in ceramide levels (e.g. C24:0-ceramide) during the transition, an increase that paralleled NEFA supply. We also observed biphasic changes in circulating sphingomyelin (e.g. C18:1-sphingomyelin levels in plasma reached a nadir at calving). These results support our hypothesis that overnutrition 1) supports impaired fatty acid oxidation and the de novo synthesis of ceramide, and 2) inflammation supports the hydrolysis of sphingomyelin to form ceramide. In further support, select ceramide levels were higher in overweight cows (e.g. C24:-ceramide) while sphingomyelin levels were higher in lean cows. To determine the origin of plasma ceramide, we profiled the concentrations of ceramide in liver, adipose, and muscle tissues pre- and postpartum. The data has been collected and the results are currently being statistically analyzed. B. Because we observed parallel increases in plasma NEFA and ceramide, we wanted to demonstrate that excess NEFA in overweight cows can support the de novo synthesis of ceramide. In collaboration with Dr. Yves Boisclair (Cornell University), we had the opportunity to acquire samples from Holstein cows infused with a soybean triglyceride (TAG) emulsion (Intralipid) for 16 h. The infusion of TAG increased circulating NEFA and hepatic lipid deposition. Within 3 h of infusion, plasma ceramide (e.g. C24:0-ceramide) levels increased. A response shared by many ceramide and glycosylated ceramide subspecies. We did not observe dramatic changes in plasma sphingomyelin levels. These results suggest that acute changes in ceramide availability can support de novo ceramide synthesis but not sphingomyelin hydrolysis. Because these cows were non-pregnant, non-lactating Holstein cows, inflammation was likely not present. To determine the origin of plasma ceramide, we profiled the concentrations of ceramide in liver and adipose collected from these cows. The data has been collected; however, the results are currently being statistically analyzed. 2. To address objective 2, we have refined existing protocols and are currently culturing primary bovine adipocytes. Because this work has just started, we have nothing to report. After Year 1, we have identified approximately 50 new biomarkers (mainly ceramides and fatty acylcarnitines) associated with the development of insulin resistance in periparturient dairy cows. On-going work is defining the relationship between these biomarkers and the mechanisms that mediate insulin resistance.
Publications
- Type:
Journal Articles
Status:
Under Review
Year Published:
2015
Citation:
Rico, J. E., V. V. R. Bandaru, J. M. Dorskind, N. J. Haughey, and J. W. McFadden. 2015. Plasma ceramides are elevated in overweight Holstein dairy cows experiencing greater lipolysis and insulin resistance during the transition from late pregnancy to early lactation. J. Dairy Sci. [under review]
- Type:
Other
Status:
Published
Year Published:
2014
Citation:
McFadden, J. W. 2014. Metabolomics: Can it advance detection of metabolic disease? Progressive Dairyman, Issue 17, October 19, 2014, pg. 17.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
Rico, J. E. and J. W. McFadden. 2014. Overconditioned prepartum cows exhibit a greater magnitude of insulin resistance and mobilize more NEFA earlier compared with lean cows. J. Dairy Sci. 97(E-Suppl. 1):335. (Abstr. 670)
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
Rico, J. E. and J. W. McFadden. 2014. Identifying biomarkers for pre-onset insulin resistance using mass spectrometry-based metabolomics: Plasma ceramides are elevated in overconditioned transition dairy cows. J. Dairy Sci. 97(E-Suppl. 1):336. (Abstr. 671)
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
Rico, J. E. and J. W. McFadden. 2014. Plasma sphingomyelins are correlated with severity of insulin resistance, and relate to plasma ceramides in overconditioned transition dairy cows. J. Dairy Sci. 97(E-Suppl. 1). (Late-Breaking Abstr. #8)
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2015
Citation:
Rico, J. E., S. Saed Samii, A. T. Mathews, and J. W. McFadden. 2015. Temporal changes in plasma sphingolipids during the transition from pregnancy to lactation in Holstein cows. J. Dairy Sci. 98(E-Suppl. 1). (Abstr. 474)
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2015
Citation:
Rico, J. E., L. S. Caixeta, Y. R. Boisclair, and J. W. McFadden. 2015. Increased plasma NEFA lowers the ratio of sphingomyelin to ceramide in Holstein cows. J. Dairy Sci. 98(E-Suppl. 1). (Abstr. W363)
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2015
Citation:
Rico, J. E., R. E. Cokeley, and J. W. McFadden. 2015. Plasma long-chain acylcarnitines are elevated in overweight dairy cows experiencing greater lipolysis and insulin resistance during late pregnancy. J. Dairy Sci. 98(E-Suppl. 1). (Abstr. W366)
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Rico, J. E. and J. W. McFadden. 2014. Plasma ceramides are elevated, and correlate with increased lipolysis and insulin resistance in an overconditioned transition cow model. First annual Navigating Lipid Research in Baltimore: Cell to system. Carnegie Institution for Science, Baltimore, MD.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Rico, J. E. and J. W. McFadden. 2014. Plasma ceramides are elevated in overconditioned transition cows, and correlate with increased lipolysis and severity of insulin resistance. West Virginia University Davis College Graduate Research Conference. Ph.D. poster competition.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Orndorff, C., J. E. Rico, Saed S. Samii, A. T. Mathews, A. Davis, and J. W. McFadden. 2014. Overweight transition dairy cows mobilize more adipose and have decreased insulin sensitivity compared with lean cows. West Virginia University Undergraduate Research Conference. Poster competition.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Davis, A., J. E. Rico, S. Saed Samii, A. T. Mathews, C. Orndorff, and J.W. McFadden. 2014. Identification of novel biomarkers for insulin resistance using mass spectrometry-based metabolomics in a dairy cow model. West Virginia IDeA Network for Biomedical Research Excellence Research Symposium. Oral talk competition.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2015
Citation:
Saed Samii, S., J. E. Rico, A. T. Mathews, C. L. Orndorff, A. N. Davis, and J. W. McFadden. 2015. Comparison of the RQUICKI estimate of insulin sensitivity with glucose and insulin tolerance in periparturient dairy cows. J. Dairy Sci. 98(E-Suppl. 1). (Abstr. M385)
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2015
Citation:
Saed Samii, S., J. E. Rico, A. T. Mathews, and J. W. McFadden. 2015. Ratio between plasma sphingolipids reveals acyl-chain specific changes during the transition from pregnancy to lactation in Holstein cows. J. Dairy Sci. 98(E-Suppl. 1). (Abstr. M413)
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2015
Citation:
Rico, J. E., L. S. Caixeta, Y. R. Boisclair, and J. W. McFadden. 2015. Increased NEFA availability promotes plasma ceramide accumulation in Holstein cows. J. Dairy Sci. 98(E-Suppl. 1). (Abstr. 472)
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2015
Citation:
Rico, J. E., L. S. Caixeta, Y. R. Boisclair, and J. W. McFadden. 2015. An acute increase in circulating NEFA does not lower total plasma sphingomyelin levels in Holstein cows. J. Dairy Sci. 98(E-Suppl. 1). (Abstr. 473)
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