Annual/Termination Reports:

[06/16/2005] [06/26/2006] [06/19/2007] [05/30/2008] [06/12/2009]

Report Information

Participants

Adams, Sean (sadams@amylin.com) - Amylin Pharmaceuticals, Inc., San Diego, CA; Azain, Michael (mazain@uga.edu) - University of Georgia; Beitz, Donald (dcbeitz@iastate.edu) - Iowa State University; Bergen, Werner (bergewg@auburn.edu) - Auburn University; Buskirk, Daniel (buskirk@msu.edu) - Michigan State University; Carey, Gale (gale.carey@unh.edu) - University of New Hampshire; Hans, Chetan (grad student) - Mississippi State University; Hargrave, Kim (grad student) - University of Nebraska-Lincoln; Hausman, Gary (ghausman@saa.ars.usda.gov) - USDA, Athens, GA; Kinder, James (kinder.15@osu.edu) - Administrator for Project, The Ohio State University; Miller, Larry (lmiller@csrees.usda.gov) - Administrator, USDA-CSREES; Miner, Jess (jminer1@unl.edu) - University of Nebraska-Lincoln; Montscioulis (grad student) - University of New Hampshire; Novakofski, Jan (jnova@uiuc.edu) - University of Illinois; Odle, Jack (jack_odle@ncsu.edu) - North Carolina State University; Rule, Daniel (dcrule@uwyo.edu) - University of Wyoming; Sainz, Roberto (rdsainz@ucdavis.edu) - University of California-Davis; Smith, Terry (trsmith@ads.msstate.edu) - Mississippi State University; Spurlock, Michael (spurloc0@purdue.edu) - Purdue University

Brief Summary of Minutes

The annual meeting was held in San Diego, CA on April 1st, 2005. Members in attendance were: Don Beitz, Jack Odle, Gale Carey, Jess Miner, Terry Smith, Jan Novokofski, Mike Spurlock, Werner Bergen, Dan Buskirk, Gary Hausman (USDA), Dan Rule, Mike Azain, Bob Sainz, Larry Miller (USDA-CSREES) and Jim Kinder (Admin). Sean Adams (Amylin Pharmaceuticals) participated as an industry guest and has been voted into the group as a sitting member.

1. Dr. Beitz opened the meeting by having brief introductions of participants.

2. The minutes of the 2004 meeting were reviewed briefly and accepted. Mike Spurlock was elected secretary.

3. Larry Miller provided an update on the USDA budget and there was considerable discussion as to the impact of proposed budget on research funding. Of particular concern was the impact of the proposed reduction (and potential elimination) of Hatch funding and increase in the indirect costs allocation.

4. The 2006 meeting was discussed, and the consensus opinion was to keep the meeting on the front end of the Experimental Biology meetings (April 1-5, 2006). The 2006 NCR-97 meeting will be held in San Francisco, on March 31st. There was a brief discussion of having the meeting on the UC-Davis campus, but this was abandoned due to the transit time and difficulty of transporting individuals to and from the UC-Davis campus.

5. Individual research reports were presented.

6. At the conclusion of the meeting, there was brief discussion of the value of extending the meeting another half day to allow more discussion vs. making more efficient use of time. This discussion has been extended via email communications and will be resolved prior to making arrangements for the 2006 meeting.

Accomplishments

1. Influence of growth stage on caspase-3, -8, and -9 activity.<br /> Two apoptotic pathways have been demonstrated--receptor mediated (caspase-8) and mitochondria initiated (caspase-9) pathways. Both pathways activate a downstream or executioner caspase-3. The goal of the current work was to determine activity of these pathways during normal growth and aging of skeletal muscle. Three ages of chickens were selected to represent the stages of rapidly growing juveniles (1 week old, n = 6), young adult (2 months old, n = 6) and aged adult (2 years old, n = 6). Muscle growth occurred in three patterns--continued growth with age (pectoralis, biceps femoris), plateau at 2 months of age (sartorius, gastrocnemius), or decrease after 2 month of age (anterior latisimus dorsi, biceps brachii). Similarly, muscle caspase activity either plateaus at 2 months of age (i.e., sartorius, gastrocnemius) or declines from 2 months of age to 2 years old (i.e., pectoralis, biceps brachii, biceps femoris). However, there was no correlation between change of caspase-3 activities and change of muscle absolute wet weight. A strong correlation between activity of the two initiator caspases (-8 and -9) implies substantial cross talk between these pathways.<br /> <br /> 2. Mechanism of regulation of adipose tissue accretion by conjugated linoleic acid.<br /> Dietary conjugated linoleic acid (CLA)-induced body fat loss is enhanced in mice fed coconut oil diets for 6 wk following weaning compared with mice fed soy oil. We utilized several strategies to test the role of the essential fatty acid deficiency caused by coconut oil in the enhanced CLA-induced body fat loss. Results indicated that coconut oil enhances the CLA effect on decreasing body fat independent of an essential fatty acid deficiency. CLA metabolites contribute to the induction of a body fat loss; however, inhibition of D6 desaturase tended to decrease the CLA-induced body fat loss (P = 0.08). The altered fatty acid profile of coconut oil- and fat free-fed mice may alter the rates of fatty acid turnover as medium-chain fatty acids are oxidized more rapidly than are long-chain fatty acids. Coconut oil- and fat free-fed mice did not have altered rates of lipolysis. CLA increased lipolysis but only in coconut oil- and fat free-fed mice and not in soy oil-fed mice. Therefore, the enhancement of CLA-induced body fat loss by coconut oil and fat-free diets seems to involve increased lipolysis. In addition to a body fat loss, CLA also increases DNA fragmentation, indicative of apoptosis, in adipose tissue. These results confirm our previous reports of increased apoptosis in the adipose tissue of CLA-fed mice.<br /> <br /> 3. Lipid supplementation of lactating beef cows: A strategy for optimizing the post partum interval.<br /> Critical factors for optimizing the post partum interval, or the successful rebreeding of beef cows, include optimal nutrition, adequate body condition, and positive growth. The objectives of the current studies were to supplement lactating beef cows with high-oleate or high-linoleate safflower seeds and determine changes in body condition, adipose tissue lipogenesis, lactation measurements including milk lipid fatty acid composition, as well as blood metabolites and hormones. No diet effects on measures of in vitro lipogenesis in adipose tissue were observed. Fatty acid profiles of the milk fat revealed dietary fat-induced decreases (P < 0.0001) in concentrations of 10:0, 12:0, 14:0, and 16:0. This result may indicate down-regulation of lipogenesis in mammary tissue. Evaluation of mRNA for acetyl-CoA carboxylase, lipoprotein lipase, fatty acid synthase, and stearoyl-CoA desaturase revealed no major changes in their abundance when control cows and linoleate-supplemented cows were compared. Overall, the dietary supplementation of high-oleate or high-linoleate oil to lactating beef cows did not affect energy repartitioning into adipose tissue from 30 to 60 days of lactation (postpartum). Future investigations will focus on more potent inhibitors of milk fat synthesis in mammary tissue as a means of allowing a portion of dietary energy to be diverted back to adipose tissue of the beef cows to provide a more rapid gain of body condition before the critical 80 day postpartum interval occurs.<br /> <br /> 4. Ontogeny of carnitine biosynthesis in pigs as inferred from gamma-butyrobetaine hydroxylase activity.<br /> Gamma-butyrobetaine hydroxylase (EC 1.14.11.1) is the last enzyme of the L-carnitine biosynthesis pathway and catalyzes the hydroxylation of gamma-butyrobetaine to L-carnitine. Apparent enzyme kinetic constants (Vmax and Km for gamma-butyrobetaine) were measured in liver and kidney homogenates from pigs in seven age categories: newborn, 24-h-old (unsuckled), 1-, 3-, 5-, 8 wk-old and adult. Total enzyme activity increased by 130 fold for liver and 18 fold for kidney as organ weight increased from birth to 8 wk. Results were that age (developmental stage) affects gamma-butyrobetaine hydroxylase specific activity and Km for gamma-butyrobetaine in liver and kidney. While the predominant organ for carnitine synthesis is likely the kidney in neonates, the liver seems to predominate after the pig exceeds 1 wk of age.<br /> <br /> 5. The effect of ractopamine feeding level on fatty acid profiles in belly fat and backfat of finishing pigs.<br /> One hundred fifty pigs (75 barrows and 75 gilts, initial weight 77 kg) were used to investigate the effect of ractopamine (RAC) on fatty acid profile in belly fat and backfat. Pigs, within genders, were assigned randomly to three treatments, which consisted of 0 (control diet) or 5 or 10 ppm of RAC. Feeding RAC had no effects on fatty acid profile and the calculated iodine value of belly fat, but had an effect on linoleic acid content in backfat. No significant difference was detected between pigs fed control diet and pigs fed control diet with 5 ppm RAC. Feeding a high level of RAC (> 5 ppm) will increase the enrichment of linoleic acid in backfat, but there is no overall effect (or negligible effect) of RAC (less or equal to 10 ppm) on the fatty acid profile in belly fat and backfat of finishing pigs.<br /> <br /> 6. Enhancement of bovine preadipocytes by troglitazone and dexamethasone.<br /> It was hypothesized that bovine i.m. and s.c. preadipocyte differentiation would be enhanced by a peroxisome proliferators-activated receptor (PPAR)-³ agonist, troglitazone (TRO), and a glucocorticoid, dexamethasone (DEX), with the relative response being greater in i.m. than in s.c. cells. In the first set of experiments, preadipocytes from i.m. and s.c. stromal-vascular (S-V) cells of an Angus steer were cloned and used to optimize culture conditions supporting adipose differentiation, as well as compare the adipogenic responses of i.m. and s.c. preadipocytes to TRO. When i.m. (n = 3) and s.c. (n = 2) clones were compared, all clones responded to addition of 20 to 60 ¼M TRO (P < 0.02), with no depot differences (P = 0.47). In the second set of experiments, i.m. and s.c. S-V cells isolated from three Angus steers were used to determine the adipogenic effects of DEX and TRO. Differentiation of bovine i.m. and s.c. preadipocytes was enhanced in response to serum lipid, DEX, and TRO. Under identical media conditions, s.c. preadipocytes have a greater capacity to differentiate compared with i.m.<br /> <br /> 7. Regulation of gene expression in porcine skeletal muscle by ractopamine.<br /> Expression of selected genes of carbohydrate and lipid metabolism was determined in skeletal muscles (ST) of Pietrain crossbred pigs after feeding of ractopamine for 52 days. Foxo 1a, PPAR alpha, lipoprotein lipase (LPL), PDK4, elF-4EBP, and beta actin mRNA abundance were studied by utilizing quantitative RT-PCR (real time). mRNA abundance for only LPL was lower (P < 0.01) in ST muscle of ractopamine-fed pigs; expression of other genes showed no differences between treatment and control.<br /> <br /> 8. Role of vasculature of adipose tissue in clearance of plasma lipids.<br /> Liposyn (artificial lipid preparation) was injected into the blood of 70- and 110-day fetal pigs to study clearance from blood and uptake by capillaries and other fetal tissues. In fetuses injected with liposyn, numerous lipid droplets were evident in adipose tissue capillaries that were not associated with adipocytes. In contrast, little lipid was evident in muscle capillaries. Electron microscopy demonstrated phagocytosis of liposyn by capillary walls and lipid droplets inside and outside capillary walls. Less lipid was evident in older fetuses injected with the higher dose of liposyn, but lipid droplets were evident in capillary walls and many other ultrastructural features were associated with liposyn infusion. Therefore, the inherent capability of fetal adipose tissue to clear lipid from blood may reside in the vasculature per se.<br /> <br /> 9. The effect of dietary omega-3 fatty acids on sow and litter performance.<br /> The objective of this study was to determine the effects of feeding a diet containing n-3 fatty acids during late gestation and/or lactation on sow reproductive performance. Omega-3 fatty acids, supplemented in the form of a protected n-3 product (Fertilium®, United Feeds, Sheridan, IN), resulted in a shift of the n-6/n-3 ratio from approximately 20 in the control to 13 in the omega-3 gestation diets and 13 to 10, respectively, in the diets. During lactation, sows fed the control diet during gestation had greater feed consumption than did those fed the omega-3 diet. Pigs from sows fed omega-3 L diet, challenged with lipolysaccharide at d 14, showed a trend for attenuated temperature change and body weight loss. Additionally, those sows fed the control diet during gestation weaned heavier litters than did sows fed the omega-3 diet. Evidently, feeding a diet containing supplemental omega-3 fatty acids during late gestation had a negative effect on sow feed intake and thus litter performance during lactation.<br /> <br /> 10. Conjugated linoleic acid (CLA) appearance in human breast milk.<br /> The purpose of this study was to determine the kinetics of CLA appearance in the breast milk of lactating mothers following ingestion of a shortbread cookie made with either CLA-enriched or CLA-moderate butter (control). Seven women expressed an average of 8 samples of breast milk over 48 hours after eating either CLA-enriched (1912 mg CLA) or CLA-moderate (231 mg CLA) cookies. Results are (1) CLA enrichment of total fatty acids in the breast milk post-ingestion of the CLA-enriched cookies was 5-fold compared with control, (2) the area under the curve for 48 hours post CLA-enriched cookies ingestion was 46% greater than that for post CLA-moderate cookies ingestion, (3) after ingestion of the CLA-enriched cookies, breast milk CLA enrichment peaked between 8-28 hours. In conclusion, breast milk fatty acids are enriched in CLA by the ingestion of a CLA-rich food product within 28 hours.<br /> <br /> <br /> 11. Adipose tissue and inflammation.<br /> Inflammation has been linked to chronic diseases such as cardiovascular disease and diabetes, and adipose tissue has been implicated as a producer of a variety of inflammatory factors. To evaluate 3-month-old miniature swine as a model for the study of inflammation and disease, we determined adipose tissue production of monocyte chemoattractant protein-1 (MCP-1) in situ, ex vivo, and in vitro. One sibling of each pair was overfed a high fat diet (HF), whereas the remaining sibling was normal-fed the control diet (C) for 3 months. No MCP-1 production was detected in adipose perfusate of any of the six swine. MCP-1 was undetectable in adipose tissue incubation fluid from C swine but increased by 76% in fluid from HF swine. Adipocytes were isolated from adipose tissue: a 2-hour incubation increased MCP-1 by 300% and 48% in adipocytes from C and HF swine, respectively. In conclusion, MCP-1 is produced by adipose tissue and adipocytes in vitro but is not detectable in situ. There was no correlation between MCP-1 production and adiposity; thus, the utility of the swine to be used as a model for studying inflammation and chronic disease is uncertain.<br /> <br /> 12. Association of genetic variation to healthfulness of beef.<br /> The objective was to determine the natural variability in fatty acid composition of beef lipids and to identify single nucleotide polymorphisms (SNPs) in the stearoyl-CoA desaturase (SCD) gene to test the association of SNPs with fatty acid composition. In general, we found that triacylglycerol (TAG) composition is heritable but phospholipids composition is not. The atherogenic index of TAGs had a heritability estimate of 0.55 and 0.45 for the TAG and total lipids, respectively. Individual fatty acids of TAGs also had high heritability estimates. For example, the heritability estimates of 14:0 and 16:0 in TAG were 0.49 and 0.40, respectively. Monounsaturates 16:1 and 18:1 in TAGs both had heritability estimates greater than 0.5. Two of the three potential SNPs for the SCD gene were homozygous in the tested population. We have classified 172 cattle into two genotypes, VA and VV, based on the third SNP. The ratio of palmitoleic acid (C16:1) concentration vs. palmitic acid (C16:0) concentration of TAG is significantly associated with the SNP (P=0.02). There is no significant association of the SNP with the fatty acid composition of phospholipids.<br /> <br /> 13. Isolation and proliferation of fish muscle satellite cells.<br /> The overall scope of this project was to screen a wide variety of relevant, growth factors and/or hormones in (fast growthrainbow trout and slow growthyellow perch) fish-derived satellite cell cultures to assess potency in prompting cell proliferation. By using collagenase, pronase, only slight centrifugation steps, and two different filtration steps, we were capable of isolating rather large numbers of satellite cells from both muscles of rainbow trout and yellow perch. Previous research indicated that trout satellite cells could be maintained at 20 ºC in non-bicarbonate-buffered L-15 medium that is supplemented with serum. This same formulation, however, did not support the proliferation of rainbow trout (or yellow perch) satellite cells. Investigators are now in the process of adding specific growth regulators [IGF-I, GH, FGF, HGH, IL-15, and others] to determine if these factors can induce fish satellite cells to proliferate in vitro.<br /> <br /> 14. Dedifferentiation of fish and bovine adipocytes.<br /> Mature adipocyte cultures were established from both beef animals (subcutaneous, visceral, and intermuscular fat depots) and from trout (visceral fat depot only). Parallel cultures of traditional stromal vascular cells also were established from trout. Investigators were incapable of prompting mature adipocytes from trout to dedifferentiate, but beef-derived mature adipocytes dedifferentiate and readily form adipofibroblasts. Adipofibroblasts were capable of both proliferating and differentiating to form lipid-filled adipocytes in vitro. Current efforts are under way to decipher the regulation of the dedifferentiation process, as well as the conversion of progeny adipofibroblasts to form adipocytes.<br /> <br /> 15. Simulation of the development of adipose tissue in beef cattle.<br /> A sub-model of bovine adipose development (hyperplasia and hypertrophy) for incorporation into a model of whole-animal growth was developed. Four adipose depots were represented: visceral (Fv), intramuscular (Fi), subcutaneous (Fs), and intramuscular (Fm) adipose tissue. During growth from 100 to 700 days of age, no plateau in cell numbers was discernible in the subcutaneous and intramuscular depots. In contrast, the visceral and intramuscular depots display clear maxima. However, there is some evidence of a second wave of hyperplasia, particularly in the visceral depot. For the purposes of this exercise, no provision for secondary hyperplasia has been incorporated, because most beef cattle would reach market finish prior to this occurrence. Even in the early-maturing tissues, hypertrophy continues with little sign of a plateau up to 600 days of age. As subcutaneous and intramuscular adipocytes proliferate, however, these tissues make up progressively larger proportions of the total body fat as the animal matures. The sub-model seems to adequately describe normal growth and accretion of adipose tissue in the whole body and in the four main depots. It represents a first step in the development of an analytical tool for the study of factors affecting fat deposition and distribution, but much remains to be done. As with all growth models, it is only as good as the estimates of nutrient inputs; so, full testing of the model will require definition of feed intake and energy value throughout the life of the animal. Further model developments will include incorporation of nutritional elements to the equations for adipose hyperplasia, possible differences in lipogenesis and lipolysis among depots, examination of possible mechanisms for genetic differences, and study of the effects of growth path or nutritional history.<br />

Publications

<b>2005 Publications <i>(see attachment below for 2004 publications):</b></i><br /> Behlke, K., E. Behlke, P. Robinson, J. Takacs, R. Dumitru, S. Ragsdale, P. Newsome, and J. Miner. 2005. Inhibition of methanogenesis in free living vs. protozoa-associated ruminal methanogens. J. Anim. Sci. (Midwest Section).<br /> <br /> Bergen, W.G., Mersmann, H.J. 2005. Comparative aspects of lipid metabolism: impact on contemporary research and use of animal models. J. Nutr. 135: 2499-2502.<br /> <br /> Bohan, M.M., G. Janda, L. Anderson, A. Trenkle, and D. Beitz. 2005. Effect of dietary macronutrients on appetite-related hormones in blood on body composition of lean and obese rats. FASEB J. 19:A76.<br /> <br /> Brewer, M. S., Peterson, W. J., Carr, T., Mccusker, R., and Novakofski, J. 2005. Thermal gelation properties of myofibrillar protein and gelatin combinations. J. Muscle Foods. 16(2):126-140.<br /> <br /> Burrin, D.G. and H.J. Mersmann (Eds.). Biology of Metabolism in Growing Animals. Elsevier 2005.<br /> <br /> Carey, G.B. 2005. Integrative metabolism: an interactive learning tool for biochemistry, physiology, and nutrition. FASEB J. 19:A222.<br /> <br /> Carnagey, K.M., S.M. Lonergan, E.J. Huff-Lonergan, R.L. Horst, A.H. Trenkle, and D.C. Beitz. 2005. Effect of 25-hydroxyvitamin D3 and dietary calcium on calpastatin activity in beef muscles. FASEB J. 19:A991. <br /> <br /> Carnagey, K.M., A.E. Wertz, T.J. Knight, R.L. Horst, E.J. Huff-Lonergan, A.H. Trenkle, and D.C. Beitz. 2005. Effects of orally administered 25-hydroxyvitamin D3 and vitamin E on calcium and vitamin D metabolites in plasma and on vitamin D metabolites in beef longissimus dorsi muscle. Abstr. 100 of Midwestern Section ADSA and Midwest Branch ADSA 2005 meeting.<br /> <br /> Corl, B.A., J.M. Rhoads, R.J. Harrell, A.T. Blikslager, O.T. Phillips, L.A. Gatlin, X.M. Niu and J. Odle. 2005. Rotaviral enteritis stimulates ribosomal p70 s6 kinase and increases intestinal protein synthesis in neonatal pigs. FASEB J.<br /> <br /> Cota, D.E., B.A. Corl and J. Odle. 2005. Isolation of feline stromal-vascular cells and differentiation into adipocytes. Undergraduate Research Symposium. NCSU. Aug. 5.<br /> <br /> Dodson, M.V., M.E. Fernyhough, J.L. Vierck, and G.J. Hausman. 2005. Adipocytes may not be a terminally differentiated cell type: Implications for animal production. Animal Science 80(3):239-240.<br /> <br /> Ebert, A.R., A.S. Berman, R.J. Harrell, A.M. Kessler, S.G. Cornelius and J. Odle. 2005. Vegetable proteins enhance growth of milk fed piglets, despite lower apparent ileal digestibility. J. Nutr. 135:2137-2143.<br /> <br /> Ele, J., S. Meers, M. Azain, and R. Dove. 2005. Evaluation of canola meal as an alternative plant protein source in nursery pig diets. J. Anim. Sci. 88 (Suppl 1): 334.<br /> <br /> Fernyhough, M.E., D.I. Helterline, J.L. Vierck, G.J. Hausman, R.A. Hill, and M.V. Dodson. 2005. Dedifferentiation of mature adipocytes to form adipofibroblasts: More than a possibility. Adipocytes 1:17-24.<br /> <br /> Fernyhough, M.E., J.L. Vierck, G.J. Hausman, P.S. Mir, E.K. Okine and M.V. Dodson. 2005. Primary adipocyte culture: Adipocyte purification methods may lead to a new understanding of adipose tissue growth and development. Cytotechnology 46:163-172<br /> <br /> Fernyhough, M., L. Bucci, G. Hausman, J. Antonio, J. Vierck,and M.V. Dodson. 2005. Gaining a Solid Grip on Adipogenesis. Tissue and Cell 37(4):335-338.<br /> <br /> Gatlin, L.A., M.T. See and J. Odle. 2005. Effects of chemical hydrogenation of supplemental fat on relative apparent lipid digestibility in finishing swine. <br /> J. Anim. Sci. 83:1890-1898.<br /> <br /> Hadenfeldt, T.J., K.M. Hargrave, and J. L. Miner. 2005. The interaction of dietary CLA and fat source on triglyceride turnover in adipose tissue of mice. J. Anim. Sci. (Midwest Section). <br /> <br /> Hargrave, K. M., M.J. Azain, M.G. Obukowicz, and J.L. Miner. 2005. Effect of conjugated linoleic acid and/or a specific delta6-desaturase inhibitor on body composition of mice. J. Anim. Sci. (Midwest Section). <br /> <br /> Hargrave, K.M., T.J. Hadenfeldt, M.J. Azain, and J.L. Miner, 2005. Coconut oil and fat free diets enhance conjugated linoleic acid-induced lipolysis and body fat loss in mice. FASEB J. 19:A447.<br /> <br /> Hart, H.A., M. J. Azain, G. J. Hausman, D. E. Reeves, and C. R. Barb. 2005. Failure of short term feed restriction to effect leptin secretion and subcutaneous adipose tissue expression of leptin or long form leptin receptor (Ob-r) in the prepuberal gilt. J. Anim Sci. 88 Suppl 1: 167.<br /> <br /> House, R.L., J.P. Cassady, E.J. Eisen, T.E. Eling, J.B. Collins, S.F. Grissom and J. Odle. 2005. Functional genomic characterization of the delipidative effects of trans 10, cis12 conjugated linoleic acid (t10c12 CLA) in a polygenic obese line of mice. Physiol. Genomics. 21:351-361.<br /> <br /> House, R.L, J.P. Cassady, E.J. Eisen, M.K. McIntosh, and J. Odle. 2005. Conjugated linoleic acid evokes delipidation through the regulation of genes controlling lipid metabolism in adipose and liver tissue. Obesity Reviews. 6:247-258.<br /> <br /> Kinkel, A.D., M.E. Fernyhough, D.L. Helterline, J.L. Vierck, K.S. Oberg, T.J. Vance, G.J. Hausman, R.A. Hill and M.V. Dodson. 2005. Oil red-O stains non-adipogenic cells: A precautionary note. Cytotechnology 46(1):49-56.<br /> <br /> Kokta, T.A., A.L. Strat, M.R. Papassani, J.I. Szasz, M.V. Dodson, and R.A. Hill. 2005. Regulation of lipid accumulation in 3T3-L1 cells: Effects of oleic and linoleic acids and insulin. The Endocrine Societys 87th Annual Meeting.<br /> <br /> Lin, X., R.L. House and J. Odle. 2005.Ontogeny and kinetics of carnitine palmitoyltransferase in liver and skeletal muscle of the domestic felid (Felis domestica). J. Nutr. Biochem. 16:331-338.<br /> <br /> Lin, X., P. Lyvers-Peffer, J. Woodworth and J. Odle. 2005. Ontogeny of carnitine biosynthesis in pigs, inferred from gamma-butyrobetaine hydroxylase activity. FASEB J.<br /> <br /> Lin, X., M.T. See, K.N. Wentz, J. Odle, B.A. Belstra, T.A. Armstrong, P.D. Matzat, P.J. Pincker, F.K. McKeith, M. Culbertson, W. Herring, and J. Hansen. 2005. The effect of ractopamine feeding level on fatty acid profiles in belly and clearplate fat of finishing pigs. Midwest Animal Science, DesMoines, IA.<br /> <br /> Meers, S. A., C. R. Dove, and M. J. Azain. 2006. The effect of omega-3 fatty acids on sow and litter performance. J. Animal Science 88 (Suppl. 1): 31.<br /> <br /> Mersmann, H.J., 2005. Mechanisms for modification of porcine growth by beta-adrenergic receptor agonists. In; Manipulating Pig Production X, JE Patterson, Ed. Australasian Pig Science Association, Werribee, Victoria, 3030, Australia. pp. 76-89.<br /> <br /> Mersmann, H.J. and S.B. Smith. Development of white adipose tissue metabolism. In: Biology of Metabolism in Growing Animals. D.G. Burrin and H.J. Mersmann, Eds. Elsevier, Edinburgh.pp. 275-302. 2005.<br /> <br /> Mersmann, H.J. and S.B. Smith. Adipose tissue development. IN: Encyclopedia of Meat Science. W. Jensen, C. Devine and M. Dikemann, Eds. Elsevier, Oxford. Pp.530-538. 2004.<br /> <br /> Mersmann, H.J. Body composition: chemical analysis. IN: Encyclopedia of Animal Science. W.G. Pond and A.W. Bell, Eds. Marcel Dekker, New York, NY. pp.159-162. 2005. <br /> <br /> Mersmann, H.J. Body composition: genetic influence. IN: Encyclopedia of Animal Science. W.G. Pond and A.W. Bell, Eds. Marcel Dekker, New York, NY. pp. 163-165. 2005.<br /> <br /> Mersmann, H.J. Body composition: technical options for change. IN: Encyclopedia of Animal Science. W.G. Pond and A.E. Bell, Eds. Marcel Dekker, New York, NY.pp.177-179. 2005.<br /> <br /> Mersmann, H.J. Lipids. IN: Encyclopedia of Animal Science. W.G. Pond and A.E. Bell, Eds. Marcel Dekker, New York, NY. pp. 578-581. 2005.<br /> <br /> Mersmann, H.J. Contributions to society: biomedical research models. IN: Encyclopedia of Animal Science. W.G. Pond and A.E. Bell, Eds. Marcel Dekker, New York., NY. pp.239-241. NY 2005.<br /> <br /> McNeel, R.L. and H.J. Mersmann. 2005. Low- and high-carbohydrate diets: body composition differences in rats. Obes. Res. 13: 1651-1660.<br /> <br /> Moutsiolis, A.A., D.C. Rule, C.M. Murrieta, D.E. Bauman, A.L. Lock, D.M. Barbano, and Carey, G.B. 2005. Conjugated linoleic acid appearance in human breast milk. FASEB J. 19:A436.<br /> <br /> Odle, J., P. Lyvers Peffer, X. Lin. 2005. Hepatic fatty acid oxidation and ketogenesis in young pigs. In: Biology of metabolism in growing animals. (Eds D.G. Burrin and H.J. Mersmann) Elsevier Limited, New York. pp. 221-234.<br /> <br /> Oliver, W.T., K.J. Touchette, C.S. Whisnant, J.A. Brown, S.A. Mathews, J. Odle, and R.J. Harrell. 2005. Pigs weaned from the sow at 10 days of age respond to dietary energy source of manufactured liquid diets and exogenous porcine somatotropin (pST). J. Anim. Sci. 83:1002-1009.<br /> <br /> Peffer, P.A., X. Lin and J. Odle. 2005. Hepatic ß oxidation and carnitine palmitoyltransferase I in neonatal pigs following dietary treatments of clofibric acid, isoproterenol and medium chain triglycerides . Am. J. Physiol. (Regul, Integr. Comp. Physiol.) 288:R1518-R1524.<br /> <br /> Rhoads, J.M., X.M. Niu, J. Odle, and L. Graves. 2005. Role of mTOR signaling in intestinal cell migration. J. Pediatr. Gastroent. Nutr. 41:514-515.<br /> <br /> Rhoads, J.M., B. Harrell, B. Corl, X.M. Niu, L. Gatlin, O. Phillips, A. Blikslager, and J. Odle. 2005. mTOR signaling is a component of intestinal repair in piglet rotavirus enteritis. J. Pediatr. Gastroent. Nutr. 41:514.<br /> <br /> Rhoads, J.M., X.M. Niu, B. Corl, R.J. Harrell, L.A. Gatlin, and J. Odle. 2005. Ribosomal p70s6k coordinately changes with intestinal and muscle protein synthesis rates during viral diarrhea. J. Invest. Med., S205 (meeting of Southern region of Amer. Physiol. Assoc.).<br /> <br /> Rincker,P.J., M.T. See, T.A. Armstrong, P.D. Matzat, B.A. Belstra, F.K. McKeith, L. Xi, J. Odle, M. Culbertson, W. Herring, and J. Hansen. 2005. Effects of ractopamine feeding level on carcass cutting yield and loin quality measurements. Midwest Animal Science, DesMoines, IA.<br /> <br /> See, M.T., T.A. Armstrong, P.D. Matzat, B.A. Belstra, F.K. McKeith, P.J. Rincker, L. Xi, J. Odle, M. Culbertson, W. Herring, and J. Hansen. 2005. Effect of ractopamine feeding level on growth performance and carcass composition. Midwest Animal Science, DesMoines, IA.<br /> <br /> Strouch, M.B., E.K. Jackson, Z. Mi, N.A. Metes, and Carey, G.B. (2005) Extracellular cyclic AMP-adenosine pathway in isolated adipocytes and adipose tissue. Obes. Res. (In press).<br /> <br /> Wentz, K., X. Lin, T. See and J. Odle. 2005. Effects of ractopamine on pork fatty acid composition. Undergraduate Research Symposium. NCSU. April 28.<br />

Impact Statements

1. Skeletal muscle growth relates to the apoptosis (programmed death) of the myonuclei as evidenced by correlations of apoptosis (e.g., caspase) activity to muscle growth.
2. Conjugated linoleic acid decreases accretion of adipose tissue through increased rates of apoptosis (e.g. caspase 3 activity) in adipose tissue.
3. Supplementation of the diet of lactating beef cows with high-oleate or high-linoleic oil did not affect energy partitioning into adipose tissue from 30 to 60 days postpartum, suggesting that rebreeding efficiency may not be improved by this nutritional strategy.
4. Prior to one week of age, most carnitine biosynthesis occurs in kidney of neonatal pigs but biosynthesis in liver predominates when pigs are older than one week of age.
5. Feeding ractopamine at doses of 10 ppm, not 5 ppm, enriches backfat of finishing pigs, but there is no overall effect on fatty acid profile of belly fat or back fat of finishing pigs.
6. Differentiation of bovine preadipocytes is enhanced by activation of the peroxisome proliferators-activated receptor and the glucocorticoid receptor.
7. Ractopamine feeding decreases expression of lipoprotein lipase (involved in fat accretion) but not of other selected enzymes of fat accretion.
8. Fetal adipose tissue, at least in part, removes blood lipids via phagocytosis of its vasculature.
9. Feeding supplemental omega-3 fatty acids to lactating sows decreased feed intake and thus litter growth.
10. Lactating women who consume foods enriched in conjugated fatty acids (CLA) produce CLA-enriched milk within 28 hours.
11. On the basis of secretion of monocyte chemoattractant protein-1, the usefulness of miniature swine as a model for studying inflammation and chronic disease remains uncertain.
12. Individual fatty acids of triacylglycerol but not of phospholipids, of beef lipid are highly heritable, and the ratio of palmitic to palmitoleic acid in triacylglycerol is associated with a single nucleotide polymorphism in the stearoyl-CoA desaturase gene.
13. Satellite cells can be isolated from fish muscle cells for mechanistic studies.
14. Mature bovine but not fish (trout) adipocytes were shown to dedifferentiate into adipofibroblasts and then to become lipid-filled adipocytes again.
15. A sub-model of bovine adipose tissue development was developed from a model of whole-animal growth. It adequately described normal growth (accretion) of subcutaneous, intramuscular, visceral, and intermuscular depots.

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Participants

The NCCC097 annual meeting was held again in conjunction with Experimental Biology, with most participants attending both meetings. In addition to Drs. Jim Kinder (administrative advisor), Mark Mirando (CSREES Representative), and Harry Mersmann (Distinguished Alumnus), the following individuals were present. All project directors provided research updates. Additionally, we were fortunate to have several invited guests, a number of graduate students, postdoctoral researchers, and senior level laboratory managers.;

Auburn: Werner Bergen;
California-Davis: Bob Sainz;
Georgia: Mike Azain;
Illinois: Jan Novakofski;
Iowa State: Mike Spurlock, Jeremy Davis, Nicholas Gabler, Jen W.-Daniels,Tim Stahly;
Iowa State: Don Beitz, Michele Bohan;
New Hampshire: Gale Carey, Andrea Arel,Tony Tagliaferro;
North Carolina: Jack Odle, Holly Hess, Sheila Jacobi;
USDA-ARS, California: Sean Adams, Pieter Oort;
USDA-ARS, Georgia: Gary Hausman;
Wyoming: Dan Rule,Chuck Murrieta;
Taiwan, National Taiwain University: S.T. Ding

Brief Summary of Minutes

Dr. Spurlock opened the meeting by introducing Drs. Kinder and Mirando. Dr. Kinder gave a brief administrative update, and stressed the importance of communicating the value of the group and meeting activity in annual reports and summaries. Dr. Mirando provided insight as to potential factors that might impact funding opportunities and NCCC097 activities. In particular, Dr. Mirando noted that the USDA and NIH were developing a joint project that would support the use of meat animal models for human applications.

Dr. Sean Adams, USDA-Western Human Nutrition Research Center, was nominated and elected as the new meeting Chair.

Project Directors, graduate students, and postdoctoral researchers provided research updates (summarized below).

Accomplishments

(1) Growth, Development, & Functional Aspects of Adipose Tissue: a unique adipocyte transcript was discovered using differential gene expression studies in rodent brown adipose tissue (BAT). The gene was found to be robustly expressed in rodent and human white adipose tissue (WAT) relative to other tissues, and is up-regulated during adipocyte differentiation in the 3T3-L1 cell system. The open reading frame of this gene (now termed Tusc5) encodes a region with high similarity to proteins containing an interferon-inducible transmembrane domain which could signal a role in adipocyte growth and maturation. Mouse genetics studies implicate Tusc5 in adiposity phenotypes. Recent tissue expression studies have revealed a highly-unusual pattern of co-expression in sensory nerve ganglia which contain the cell bodies of nerves carrying information from the periphery to the brain. These initial results suggest an important role for Tusc5 in normal adipocyte maturation and function, and indicate that at the protein shares a functional role between fat cells and sensory nervous system cells.<br /> <br /> (2) Dietary Constituents Regulating Lipid Deposition or Lipid Composition: (2a) The effects of feeding a diet containing omega-3 (n-3) fatty acids during late gestation (G) or lactation (L) on sow milk composition were determined. Encapsulated n-3 fatty acids (Fertilium, United Feeds, Sheridan, IN) added to corn-SBM-based G and L diets resulted in a shift of the n-6/n-3 ratio from ~20 in control sows to 10-13 in n-3 supplemented sows. The addition of n-3 increased the total n-3 percent in milk (1.35%, vs. 1.12% in controls, p<0.0001) and significantly reduced the milk n-6/n-3 ratio from 17.49 to 14.56. n-3 addition to the L diet increased the level of EPA and DHA , regardless of whether sows consumed n-3 in the G diet. Litter weights were not affected by these interventions. Thus, even small amounts of n-3 in the maternal diet can influence milk composition. (2b) The influence of ractopamine (RAC) and dietary protein on carcass composition, performance, and meat quality were evaluated in crossbred gilts and sows. Dietary treatments included feeding crude protein (CP) at 16% (16CP), 18% (18CP), or 16% plus supplementation with Lys, Met, Thr to equal the 18% CP diet (16AA), with or without RAC at 10 ppm for 28 d. There was a trend toward higher ADG vs. the other groups for pigs fed 16AA, and these animals had significantly increased 28 d F:G vs. 16CP. RAC increased 28 d G:F, and UBF and ULM were increased in RAC pigs vs. controls. Tenth rib backfat (TRBF) was lower in 18CP vs. 16CP, with 16AA intermediate. RAC-fed pigs tended (p<0.08) to have lower TRBF, and had larger lean mass areas vs. controls. No meat quality differences were observed across dietary protein level or gender; LM from RAC-fed pigs was less red and less yellow, had higher 24 h pH, and higher NPPC marbling score. Overall, RAC supplementation may be an effective means of improving composition without compromising quality, regardless of dietary protein level.<br /> <br /> (3) Regulation of Energy Balance: Male Sprague Dawley rats and Zucker fatty rats were used to test how diets differing in macronutrient content influence a suite of hormones believed to participate in appetite regulation and energy balance. Rats were assigned for 5 wk to one of five diets: control, 75% of control calories, American Heart Association (AHA), Atkins, or high fat (HF). Weekly plasma samples were analyzed for ghrelin, leptin, insulin, and adiponectin, and 5 wk plasma also assayed for glucagon, oxyntomodulin, and metabolites. Macronutrient content in the diet and genetic background altered endocrine patterns. For instance, in obese Zuckers glucagon was significantly higher in the Atkins rats compared to HF-fed or calorie-restricted animals, and tended to be higher vs. control diet or AHA rats (p<0.1). In lean rats, glucagon was highest in the control conditions relative to other diets. These results support the idea that the endocrine response to dietary macronutrient can differ in obese vs. lean animals.<br /> <br /> (4) Dietary Constituents Regulating Lipid Deposition or Lipid Composition: A porcine-specific oligo gene array platform (provided by the Pig Genome Coordination Program at Iowa State University) was used to analyze differentially-expressed RNA transcripts in various pig tissues following 28 d RAC supplementation (60 ppm) (Study XP1) or after high-fat (40% HF) feeding for 14 d (Study XP2) in finishing pigs. Treatment-related differences in transcript intensities were evaluated using algorithms designed to account for multiple-comparisons in micro arrays (SAM software, Stanford University). In Study XP1, 550 genes were upregulated, and 569 downregulated, in WAT by RAC. Almost half of these genes lacked adequate sequence information to positively identify. RAC decreased expression of fatty acid synthase, stearoyl-CoA desaturase, acetyl-CoA carboxylase, and leptin, and increased some oxidative transcripts. In Study XP2, HF-feeding was found to alter expression of 1055, 847, and 1138 genes in liver, adipose, and muscle, respectively. In liver, FABP, fatty acid synthesis genes, and fatty acid oxidation genes were decreased. In adipose, expression of FABP, leptin, and TAG synthesis genes were increased by HF, and in muscle this treatment led to upregulation of CPT1, 3-hydroxyacyl-CoA dehydrogenase, and some genes related to oxidative phosphorylation.<br /> <br /> (5) Growth, Development, & Functional Aspects of Adipose Tissue: The impact of the lipid-soluble polybrominated diphenyl ethers (PBDEs) on adipose and mammary biology was determined. Since these chemicals are widely-dispersed in the environment, exposure may be relatively high and accumulation in fatty tissues may influence normal processes of adipose development and milk composition. PDBEs are lipophilic, continuously-released from consumer products (i.e., upholstery, computers, curtains), and persistent in the environment. They are believed to disrupt several endocrine pathways, and levels in biofluids such as milk could be important considering that magnification of PDBE concentrations occurs up the food chain. A study currently underway is examining the PDBE levels of breast milk in women whose exposure is assessed by a comprehensive questionnaire regarding their home and workplace environment, to understand potential contamination and whether this is correlated with possible environmental sources. A second study has determined that 4 wk exposure to PDBEs, given by oral gavage to rats (14 mg/kg in oil) led to significantly increased isoproterenol-stimulated lipolysis, and markedly reduced insulin-stimulated glucose uptake, in adipocytes isolated from treated rats vs. controls. These effects were not observed when control adipocytes were acutely exposed to PDBEs, or in rats dosed in vivo for 2 wk, however.<br /> <br /> (6) Growth, Development, & Functional Aspects of Adipose Tissue: The ontogeny of expression for twenty secreted proteins was evaluated in stromal-vascular (SV) cells and adipose tissue derived from 105 d fetuses and young pigs (aged 5, 90, 150, and 210 d), using microarray and RT-PCR technologies. Genes assayed included apolipoprotein A-1 (APO-A1), relaxin, RANTES, brain-derived neurotrophic factor (BDNF), IGF-binding protein 5 (IGFBP-5), several interferons, and a number of interleukins. Outer (OSQ) and inner (ISQ) subcutaneous adipose were evaluated. Microarray results indicated that, with the exception of connective tissue growth factor (CTGF) which was upregulated in OSQ with age, no genes encoding secreted molecules were changed with age in pig adipose tissue. However, real-time quantitative RT-PCR indicated significant changes in IL-1A, IL-1B, IL-6, RANTES, and several others with age. Therefore, the endocrine function of WAT changes in developing pigs, which could have functional effects on adipose growth, as well as metabolic function locally and in other tissues.<br /> <br /> (7) Dietary Constituents Regulating Lipid Deposition or Lipid Composition: (7a) The effects of supraphysiologic arachidonic acid (AA) exposure to the gut via supplementation of a milk-based formula to newborn piglets were determined. Specifically, the time-related effects of AA on fatty acid composition of enterocyte phospholipids and on the abundance of delta6- and delta5-desaturase mRNAs were tested. The diets contained either no PUFA (control), 5% eicosapentanoic acid (EPA), or 0.5%, 2.5%, 5% AA, and were fed for 4, 8, or 16 d starting at 1 d of age. At 16 d, enterocyte phospholipids containing AA were enriched significantly (i.e., by 355% in the 5% AA group) vs. controls or EPA treated, whereas in the latter phospholipids displayed a significant increase in EPA. Diet had no effect on growth, and desaturase mRNA did not differ with the exception of a reduction in the 2.5% AA group relative to control. (7b) The degree by which carnitine supplementation to sows increases tissue carnitine concentration in pig fetuses was tested. Pregnant gilts were fed control diets or diets supplemented with carnitine (88 mg/d) during the first 70 d gestation, and tissue carnitine concentration and carnitine palmitoyltransferase (CPT) kinetics determined in fetuses at 70 d. Maternal supplementation increased fetal liver and muscle tissue carnitine concentration significantly (by 58% and 33%, respectively) but did not affect kidney levels. CPT activity and kinetics were unaffected by changes in tissue carnitine concentration.<br /> <br /> (8) Dietary Constituents Regulating Lipid Deposition or Lipid Composition: Potential dietary fat-related shifts in bovine mammary tissue mRNA levels for genes encoding proteins relevant to lipid uptake and lipogenesis were evaluated (acetyl-CoA carboxylase, fatty acid synthase, lipoprotein lipase, stearoyl-CoA desaturase). The overall objective was to employ strategic nutritional inputs (fatty acid supplements) to affect mammary lipogenesis, which may influence adipose tissue accretion and thus modulate the postpartum interval for successful rebreeding. Primiparous beef cattle were fed either a low-fat control diet or a cracked high-linoleate safflower see supplement (LIN), and mammary lipogenic gene expression was evaluated both through direct measurement of mammary tissue samples derived at slaughter, and via mRNA isolation and analysis from the milk (exuded cells). Correlation of mRNA abundance comparing tissue and milk somatic cell RNAs was significant for all genes tested (Pearson correlation coefficients of 0.67-0.90; p<0.05), indicating that milk cell RNA is a valid surrogate for mammary tissue RNA for these genes. No significant treatment-related shifts in gene expression was observed. The milk mRNA protocol was used to test whether differences in body condition score (BCS) and dietary fat alter expression. Three year old beef cows were nutritionally managed to achieve a BCS of 4 or 6, and then given a low-fat control diet or one of two high-fat supplements (linoleate or oleate). Milk somatic cell RNA was analyzed at 30 d and 60 d of lactation. Lipid supplementation had no effect on gene expression; however, lower BCS was characterized by increased FAS and SCD expression, possibly reflective of a greater requirement for de novo mammary lipogenesis to compensate for the lower overall adipose to draw on for milk fat synthesis. There were significant effects of lactation duration on gene expression, with increased LPL and reduced FAS at 60 d vs. 30 d, which might reflect a shift toward higher demand for mammary uptake of circulating lipids and lower lipid synthesis later in lactation.<br /> <br /> (9) Growth, Development, & Functional Aspects of Adipose Tissue: A dynamic, mechanistic model of beef cattle growth and body composition is being developed, with a goal to provide a predictive model useful to industry (as a decision-aid tool) and researchers (to test hypotheses and interpret results). The current Davis Growth Model evolved from previous models and is based on the concepts of hyperplasia and hypertrophy to simulate the accretion of whole body protein and four fat depots (visceral, intermuscular, subcutaneous, and intramuscular). A meta-analysis of over 20 yr of literature has been performed to aid in model development. Reference animals growing from 120-600 kg have been established, with accompanying compositional data: early-moderate maturing and late maturing steers, both implanted and non-implanted. Preliminary fits using the model equations for fat distribution indicate little variation in parameter values for the various scenarios, indicating that the model responds well to variations in mature size and endocrine status.<br /> <br /> (10) Growth, Development, & Functional Aspects of Adipose Tissue: The autocrine/paracrine regulation of glucose transport and fatty acid oxidation by adiponectin has been studied in primary pig adipocytes and in the 3T3-L1 line. Adiponectin stimulates the expression of several genes readily identifiable with fatty acid oxidation, and in fact, stimulates palmitate oxidation in vitro. Additionally, the potential for hyperglycemic conditions to induce insulin resistance in adipocytes was investigated. Insulin resistance (decreased insulin-mediated 2-deoxy-D-glucose uptake) was induced by hyperglycemia, but adiponectin did not alleviate this condition. However, adiponectin did suppress the induction of reactive oxygen species (ROS) by hyperglycemia, an effect that was achieved in part by suppression of NADH oxidase and PKC-´. The regulation of ROS may be an key mechanism by which adiponectin suppresses the inflammation associated with hyperglycemia.<br /> <br /> (11) Growth, Development, & Functional Aspects of Adipose Tissue: The possibility that mature fat cells can return to a proliferation-competent state and to de-differentiate under certain conditions is being examined through the use of adipofibroblasts (progeny from de-differentiated adipocytes) in culture. Regulation of lipid metabolism, adipogenesis, and plasticity (ability to convert to other cell types) are being studied in this cell model to characterize the physiological characteristics of de-differentated fat cells.

Publications

Adams, S.H., C. Lei, C.M. Jodka, S.E. Nikoulina, J.A. Hoyt, B. Gedulin, C.M. Mack, and E.S. Kendall. 2006. PYY[3-36] administration decreases the respiratory quotient and reduces adiposity in diet-induced obese mice. J. Nutrition, 136: 195-201.<br /> <br /> Adams, V.L., C.D. Gilbert, H.J. Mersmann, and S.B. Smith. 2005. Conjugated linoleic acid depresses [3H]-thymidine incorporation into stromal-vascular cells of adipose tissue from postweanling pigs. Adipocytes 1:65-72.<br /> <br /> Azain, M. J., J. R. Broderson, and R. J. Martin. 2006. Effect of long-term somatotropin treatment on body composition and lifespan in aging obese Zucker rats. Experimental Biology and Medicine. 231: 76-83.<br /> <br /> Brewer, M.S., J. Novakofski and K. Freise.2006. Instrumental evaluation of pH effects on ability of pork chops to bloom. Meat Science 72( 4):596-602.<br /> <br /> Fernyhough, M.E., J.L. Vierck and M.V. Dodson. 2006. Assessing a non-traditional view of adipogenesis: adipocyte dedifferentiation  mountains or molehills? Cells, Tissues, Organs 182(3-4): [accepted 10 April]<br /> <br /> Jablonski, E. A., R. D. Jones, and M.J. Azain. 2006. Evaluation of pet food by-product as an alternative feedstuff in weanling pig diets. J. Anim. Sci. 84: 221-228.<br /> <br /> Novakofski, J. and M. S. Brewer. 2006. The Paradox of Toughening During The Aging of Tender Steaks, Journal of Food Science. In press.<br /> <br /> Schlegel, M.L., Bergen, W.G., Schroeder, A.L., VandeHaar, M.J., Rust, S.R. 2006. Use of bovine somatotropin for increased skeletal and lean tissue growth of Holstein steers. J. Anim. Sci. 84:1176-1187.<br /> <br /> Strat, A.L., T.J. Kokta, M.V. Dodson, A. Gertler, Z. Wu and R.A. Hill. 2005. Early signaling interactions between the insulin and leptin pathways in bovine myogenic cells. Biochemica et Biophysica ACTA (Molecular Cell Research) 1744(2):164-175.<br /> <br /> Wang, H.C., Y.K. Ko, H.J. Mersmann, C.L. Chen and S.T. Ding. 2006. The expression of genes related to adipocyte differentiation in pigs. J. Anim. Sci. 84:1059-1066. <br />

Impact Statements

1. A new adipocyte gene, Tusc5, has been identified and initial characterization in human and rodent samples suggests a potential functional role in both fat cells and cells of the sensory nervous system.
2. Addition of small amounts of an omega-3 supplement to a lactation diet significantly alters the composition of sows milk, as reflected in a significantly reduced n-6/n-3 ratio, and an increase in DHA and EPA content.
3. Ractopamine supplementation at 10 ppm improved carcass composition without compromising quality, regardless of gender or dietary protein level (16% or 18% CP).
4. The patterns of hormones relevant to metabolism and appetite in response to differences in dietary macronutrient composition or calorie restriction differed between obese and non-obese rats, suggesting that adiposity influences the regulation of nutrient-responsive endocrine pathways.
5. A pig-specific microarray has proven useful to detect tissue transcript changes in response to ractopamine and high-fat feeding, i.e., to determine that ractopamine downregulates lipogenic genes and leptin expression in porcine adipose tissue.
6. Polybrominated diphenyl ethers (PDBEs) are ubiquitous environmental toxins which accumulate in biofluids, and have been found to promote adipocyte insulin resistance and enhanced stimulation of lipolysis by isoproterenol.
7. The expression pattern of a suite of secreted proteins has, for the first time, been determined in fetal and growing pig adipose tissue, (a) indicating that potentially-important shifts in expression occur with age and could influence adipose growth and metabolism, and (b) highlighting the endocrine nature of porcine adipose tissue.
8. Supraphysiologic exposure to arachidonic acid (AA, ³ 0.5% of fatty acids in a milk-based formula) in the newborn piglet model increases enterocyte phospholipid AA content without affecting growth rate or enterocyte desaturase mRNA abundance.
9. The tissues of fetal piglets may be enriched with carnitine through supplementation of the gestating sow, and this does not influence the kinetics of CPT in isolated tissue.
10. A new method to isolate mRNA from bovine milk somatic cell pellets was used to determine that, at least for the four lipid-relevant transcripts tested, this preparation serves as an excellent surrogate for mammary tissue proper in terms of monitoring gene expression changes in response to fat supplementation in cows.
11. Preliminary analyses indicate that a beef cattle growth and body composition predictive model, based on meta-analysis of the data contained in 43 papers, responds well to variations in mature size and endocrine status of growing animals in mathematical simulations.
12. Experiments to date indicate that adiponectin does impact energy metabolism (and possibly feed efficiency), and identifies novel targets for genetic or pharmacological manipulation.
13. A growing body of evidence indicates that at least some mature fat cells can de-differentiate to form adipofibroblasts, and studies are underway to further characterize the physiology of these cells since their formation in vivo may have important implications for the animal industry (altering carcass composition) and in human health (modifying obesity and diabetes outcomes).
14. Collaborative Efforts:<ol><li>Bergen/Odle: submitted USDA-CREES-NRI proposal related to use of pig genome microarray to examine differentially-expressed genes in various porcine experimental models. <li>Adams/Spurlock: initiated collaboration as Co-Investigators examining adipose tissue physiology during weight loss (National Dairy Council grant, M. van Loan, PI). <li>Dodson/Hausman: Drs. Dodson and Hausman have worked on projects related to adipose tissue growth and differentiation, resulting in 8 research articles published or in preparation.</ol>

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Brief Summary of Minutes

Accomplishments

Accomplishments:<br /> <br /> The research objectives for NCCC097, October 2004 to September 2009, are:<br /> (see http://nimss.umd.edu/homepages/outline.cfm?trackID=3848)<br /> 1. Share and critique new techniques, experimental designs, and new unpublished data regarding adipocyte biology. <br /> 2. Through the interactive process at the meeting and afterward, elicit input to improve experimental design and to develop mutual interests toward joint projects. <br /> 3. Study biological regulation of catabolic (fatty acid mobilization and oxidation) and anabolic (fatty acid synthesis, elongation and desaturation, and triacylglycerol synthesis) lipid metabolism in domestic species at the molecular, cellular, tissue, and organismal level <br /> 4. Study cellular and molecular controls of adipocyte hyperplasia, differentiation and growth in domestic species. <br /> 5. Study nutrient determinants of excess fat deposition (diet composition, and amount and source of dietary fat, calories, and protein). <br /> 6. Study mechanisms associated with genetic variation in excess fat deposition and the post-genomic variation of individual phenotypes (determinants of cellularity, receptor populations, alternative metabolic signalling pathways, alternative gene expression and promoter function) <br /> 7. Establish approaches to maintain or enhance intramuscular fat concentration, but at the same time maintain or decrease subcutaneous and intermuscular fat. Intramuscular fat (marbling) is an important contributor to the organoleptic properties of meat, but the molecular and cellular mechanisms that retard intramuscular fat deposition until late in development are not understood. <br /> 8. Investigate the interplay between adipose tissue, liver, gut, and skeletal muscle lipid metabolism. The growth of adipose tissue is regulated, not only within the adipocyte, but also by the the interorgan interchange of metabolites, various lipoproteins, and numerous hormones and signalling molecules. <br /> 9. Investigate the role of endocrine factors produced by the adipocyte in co-ordinate regulation of feed intake, reproductive competency, energy expenditure, and immune function. <br /> 10. Study mechanisms for intervention strategies to decrease excess fat deposition (exercise, caloric restriction, pharmacotherapy, selective control of gene expression through transcription factors and modulation of promoter function). <br /> Given these objectives, NCCC097 scientists conducted the following work over the past year. The Experiment Station, followed by the objective number(s) met by the work, are provided for each entry.<br /> <br /> Growth, Development & Functional Aspects of Adipose Tissue<br /> <br /> 1. Experiment Station: Washington State University; Project objectives: 4, 6. Bovine adipocytes and have been isolated, purified and initially studied in vitro to determine the phenomenon of mature fat cells returning to a proliferative-competent state (dedifferentiation). Present research has stalled, while determination of a specific mechanism (other than employment of metabolic markers) that might be used to discern cellular switching events in the dedifferentiation of mature adipocytes is investigated. Further, single cell cytomic, proteomic and genomic analysis is being studied, which will be needed in order to evaluate single mature cells. This research has application to both the animal industry (altering carcass composition), as well as with human health (obesity and diabetes). <br /> <br /> 2. Experiment Station: University of Georgia and USDA/ARS, Richard B. Russell Agriculture Research Center, Athens, Georgia; Project objectives: 1, 5, 9, 10. Ovariectomized (OVX) prepuberal gilts averaging 164 d of age and 79 ± 4 kg BW were fed either 3 kg of feed (control fed [CF]; n = 6) or feed restricted (RST; 33% of CF diet; n = 6) for 8 d. On d 8, blood samples were collected every 15 min for 8 h for LH and leptin assay. Real-time PCR was performed on total RNA extracted from MSQ fat collected on d 9. Total RNA was also extracted from three additional adipose tissue depots and subjected to microarray analysis for feed restriction effect. A compromise of an ANOVA normalization based method and a LOWESS normalization-based method were used analysis of microarray data. Following normalization the adjusted data is analyzed, for each gene separately, via a mixed linear model with the biological replication as a random effect. CF gilts gained more BW (8.2 vs. -2.5 kg; SE = 0.6 kg; P < 0.001) and BF (2 vs. -3 mm; SE = 1mm; P < 0.02) than RST gilts. RST failed to affect indices of leptin or LH secretion except for LH pulse amplitude which was greater (P < 0.01) than CF gilts. RST failed to affect MSQ expression of leptin, long form leptin receptor (Ob-rb), AFABP (adipocyte fatty acid binding protein), C/EBP± or PPAR³2 transcripts compared to CF gilts. In the microarray study, the number of genes significantly (P = 0.001) affected by RST study was depot dependent. Regardless of level of significance, stearoyl-CoA_desaturase, PPAR³, LPL, and malate dehydrogenase decreased the most with feed restriction while AFABP, C/EBP±, and glycerol-3-phosphate dehydrogenase changed very little.<br /> <br /> 3. Experiment Station: USDA/ARS Western Human Nutrition Research Center and University of California, Davis, Davis, California; Project objectives: 1, 6, 10. Initial characterization of tumor suppressor candidate 5 (Tusc5) was introduced at last years NCCC097 meeting, and has been extended to further described at the 2007 meeting. First, based on the presence of an IFN-stimulated growth regulator domain present in Tusc5, plus its induction by PPARg agonism and stimulation toward the end of the adipogenic differentiation cascade, it is hypothesized that the function of this protein is to promote fat cell growth arrest in the fully-differentiated phenotype. Thus, levels should be reduced in obese white adipose tissue, a site which may display elevated adipocyte turnover and growth, at least during the phase in which adiposity gain is most robust. Working with collaborators from France and the U.S., we have confirmed that Tusc5 mRNA expression is in fact significantly reduced in the WAT of obese Pima Indians and a French cohort of obese women, albeit modestly. Second, we originally observed two transmembrane domains in Tusc5, and tested whether the protein is on the cell surface of adipocytes. Preliminary results, however, point to an intracellular localization, likely with Golgi or ER. Third, the unique co-expression of Tusc5 in adipocytes and peripheral nerves has been confirmed by ISH, and we have identified other genes with similar expression. In another line of research, we are attempting to identify markers of fatty acid combustion in muscle, in order to use said markers in prognostic and diagnostic tests to evaluate an individuals propensity toward poor fat utilization in this tissuethe latter is a hallmark of insulin resistance and predicts development of type 2 diabetes. We are working with our colleagues in the metabolomics arena and with Dr. Mary-Ellen Harper of the University of Ottawa, and others, to use broad metabolite analysis toward this end. Our preliminary data indicate that in the course of fatty acid b-oxidation, many metabolites from previously-unappreciated pathways and pathways related to glucose and amino acid metabolism are affected. Current studies are focused on further confirming these results and ultimately we wish to test candidate molecules using plasma from people of differing insulin sensitivity and in response to nutritional and physical activity interventions.<br /> <br /> 4. Experiment Station: The University of Wyoming; Project objectives: 1, 3, 5, 8. Using quantitative (real-time) PCR and Western blot analysis, mRNA transcript abundance of three enzymes and two protein transcription factors involved in lipogenesis in subcutaneous (s.c.) adipose tissue during lactation in the beef cow were evaluated. The overall project objective was to employ strategic nutritional inputs (fatty acid supplements) to affect mammary gland lipogenesis such that adipose tissue accretion could occur rapidly enough for the lactating beef cow to attain an 80-day post partum interval for successful rebreeding. We hypothesized that BCS at parturition and postpartum dietary fat supplementation will alter protein transcription factors and mRNA abundance of adipose tissue lipogenic and lipolytic enzymes during lactation in beef cows. Our objective was to determine abundance of mRNA for acetyl-CoA carboxylase (ACC), hormone-sensitive lipase (HSL), and lipoprotein lipase (LPL), and protein transcription factor (STAT-5 and PPAR-g) levels in adipose tissue of 3-yr old Angus × Gelbvieh beef cows nutritionally managed to achieve a BCS of 4 ± 0.07 (BW = 479 ± 36 kg; n = 18) or 6 ± 0.07 (BW = 579 ±53 kg; n = 18) at parturition. Beginning 3 d postpartum, cows within each BCS were assigned to isonitrogenous and isocaloric diets of hay plus low-fat control (CON) supplement, or supplements (5% of DMI as fat) with either cracked high-linoleate (LIN) or cracked high-oleate (OLE) safflower seeds until d 60 of lactation. At d 30 and 60 of lactation, s.c. adipose tissue biopsies were collected for RNA extraction, quantitative RT-PCR determination of transcript abundance, and Western blot analysis for STAT-5 and PPAR- ³. Adipose tissue of BCS 4 cows had less mRNA for LPL (P = 0.001) and HSL (P = 0.09) compared with BCS 6 cows. Abundance of LPL mRNA was lower (P = 0.002) at d 30 postpartum compared with d 60; whereas, HSL mRNA was greater at d 30 (P = 0.001). Cow BCS did not affect (P = 0.35) ACC mRNA; however, it tended to be higher (P = 0.13) at d 60 compared to d 30 of lactation. Abundance of PPAR- ³ tended (P = 0.13) to be lower in adipose tissue of BCS 4 cows compared with BCS 6 cows. Both STAT-5 (P = 0.0001) and PPAR- ³ (P = 0.05) were greater at d 30 compared to d 60 postpartum. We conclude that abundance of adipose tissue mRNA for LPL and HSL are influenced by cow BCS, and changes in mRNA abundance during lactation indicates a shift in nutrient partitioning away from the mammary gland to s.c. adipose tissue. Furthermore, STAT-5 and PPAR-g likely play a role in the transcription regulation of LPL and HSL in adipose tissue during lactation in beef cows.<br /> <br /> 5. Experiment Station: University of New Hampshire; Project objectives; 1, 8, 9. The possible causal link between obesity and atopic airway disease was investigated. Allergy and asthma are type 2 acquired immune disorders that develop because of activation of CD4 T helper lymphocytes by antigen presenting cells, such as macrophages. Atopy, (a biochemical precondition of airway allergy development, marked by an elevation in circulatory IgE antibodies, to common aeroallergens) was greater among young healthy adult obese (n = 21) than non-obese (n = 22) women (p <0.01). Plasma leptin, estradiol and insulin resistance also were found to be higher among obese than non-obese women (p <0.05). Using multivariate logistic regression analysis with fat mass leptin, estradiol and insulin resistance as independent factors, only percentage fat mass was significant and positively correlated with atopy among the women. The healthy women were studied further, along with 18, obese and non-obese women with asthma of similar age, to identify a possible mechanism(s) that could explain the link between fat mass and atopy. Atopy and total circulatory levels of specific IgE antibodies were greater among obese and asthmatic women in comparison to respective controls (p < 0.05). Plasma adiponectin was lower between asthmatic and non-asthmatic women; and was lower between obese and non-obese women, but only when specific IgE antibodies was treated as a covariate. Dietary analysis of the women using a food frequency questionnaire, indicated that intake of arachidonate (AA), primarily from animal protein sources, was greater among asthmatic (p <0.01) and obese (p <0.1) than their respective controls. Using multivariate regression analysis with dietary AA, plasma adiponectin, and percent fat mass as independent factors, fat mass and AA were positive predictors of total specific IgE antibodies (p <.05). Arachidonate is precursor of the eicosanoid metabolite, prostaglandin E2 (PGE2). PGE2 is produced by macrophages and has been shown, in vitro, to modify CD4 T helper cells to produce cytokines that signal type 2 acquired immunity. Weight gain therefore could promote recruitment of macrophages, that can be modified in phenotype by the n-6 fatty acid, arachidonate to produce eicosanoids that favor development of a type 2 immune response. Miniature swine are currently being used to investigate this question.<br /> <br /> 6. Experiment Station: Michigan State University; Project objectives: 1, 3, 7, 9. Signaling mechanisms that explain the lower adipogenic capacity of bovine intramuscular (IM) preadipocytes when compared to subcutaneous (SC) preadipocytes were examined. We hypothesized that the lower adipogenesis of IM preadipocytes was caused by decreased glucocorticoid receptor expression (GR), sensitivity to glucocorticoids, peroxisome proliferator-activated receptor³2 (PPAR³2) expression, and(or) PPAR³2 ligand synthesis. Stromal-vascular cells, containing preadipocytes, were isolated from IM and SC adipose tissue of 3 Angus-cross steers. Immunoblot analysis detected GR immunoreactive bands of ~97, ~62, and ~48 kDa, which were equally expressed in IM and SC cells (P > 0.50). Intramuscular preadipocytes were less adipogenic than SC preadipocytes as determined by glycerol-3-phosphate dehydrogenase (GPDH) activity and oil red O staining (P < 0.05). Dexamethasone (DEX), a synthetic glucocorticoid, increased GPDH activity similarly in preadipocytes from both depots (P < 0.05). Immunoblot analysis revealed a PPAR³2 immunoreactive band of ~53 kDa, which was expressed equally in IM and SC cells (P = 0.39). Conversely, IM cells secreted more derivatives of the presumptive PPAR ligand prostacyclin (PGI2) than SC cells (P = 0.046). Because exposure of SC preadipocytes to an inhibitor of PGI2 synthesis had no effect on adipogenesis (P = 0.99), and exogenous cPGI2 (PGI2 analog) tended to enhance adipogenesis (P = 0.06), the greater secretion of PGI2 derivatives by IM preadipocytes does not explain their lower adipogenesis. Exposing IM and SC preadipocytes to the cyclooxygenase (COX) inhibitor/ PPAR³2 ligand, ibuprofen (IBU) for 48 h or 12 d resulted in a treatment by depot interaction (P = 0.002). Ibuprofen exposure for 48 h enhanced DEX stimulation of GPDH activity only in IM cells (P = 0.009). Exposure to 100 ¼M and 500 ¼M IBU for 12 d enhanced DEX induction of differentiation in IM preadipocytes, whereas only 100 ¼M IBU enhanced DEX induction of differentiation in SC preadipocytes (P d 0.05). In the absence of DEX, exposure to IBU for 12 d maximally increased GPDH activity in IM preadipocytes by 12-fold, but only increased GPDH activity by 1.5-fold relative to control in SC preadipocytes (P < 0.001). Contrary to IBU, 500 ¼M aspirin (a COX inhibitor) did not affect GPDH activity either alone (P > .37), or combined with DEX (P > 0.60) in either cell population. Because IBU diminished adipogenic differences between IM and SC preadipocytes, it is suggested that these adipogenic differences may be partially related to differences in the endogenous activation of PPAR³2. The use of selective PPAR³2 agonists or antagonists offers potential to selectively alter adipogenesis in economically-important bovine adipose tissue depots.<br /> <br /> 7. Experiment Station: Iowa State University; Project objectives: 1, 3, 6. The objective of this study was to identify single nucleotide polymorphisms (SNPs) in bovine fatty acid synthase (FAS) gene and to test the association of SNPs with beef fatty acid composition. Sequencing of the four exons of the thioesterase (TE) domain of FAS from Angus bulls identified three novel SNPs, A17924G, C18663T, and C18727T. Polymorphism A17924G was predicted to result in an amino acid change from threonine (genotype AA) to alanine (genotype GG). A PCR-RFLP assay was developed to genotype 331 Angus bulls for the A17924G SNP in which three genotype classes were detected, AA (n =121), AG (n = 168), and GG (n = 42). The genotypes were significantly associated with fatty acid composition of longissimus dorsi (LD) muscle of purebred Angus bulls. Cattle with the GG genotype had a lower atherogenic index (AI), less percentages of myristic acid (14:0) and total saturated fatty acid (SFA), and greater contents of oleic acid (18:1) and total monounsaturated fatty acids (MUFA) in the triacylglycerol (TAG) fraction of LD muscle than did the beef cattle with genotypes AA (P < 0.05). The differences of individual fatty acid percentage in total lipid of LD muscle were similar as those in TAG fraction. In conclusion, this SNP may be used as a DNA marker to select breeding stock to improve the healthfulness of fatty acids in beef.<br /> <br /> <br /> 8. Experiment Station: Auburn University; Project objectives: 1, 2, 3, 6, 8, 10. Animal work was initiated to assess aspects of lipogenesis, mitochondrial function and biogenesis, fatty acid oxidation (beta oxidation and peroxisomal oxidation) and the role of uncoupling proteins at the ribonomics level in back-fat and loin muscle from genetically diverse cattle with an emphasis on residual feed efficiency. To this end qRT-PCR real time methodology is being developed appropriate for transcription profiling bovine tissues.<br /> <br /> 9. Experiment Station: The Ohio State University; Project objectives: 3, 4, 6, 10. The nine currently identified cytoplasmic fatty acid binding proteins (FABPC) belong to the superfamily of lipid binding proteins. FABPC all possess similar protein structures but show tissue specific expression. This tissue specific expression pattern suggests unique or specialized functions for the various FABPC. Recently, animal genetic studies suggested H-FABP is a candidate gene responsible for intramuscular fat accumulation in pigs. We investigated the expression of H-FABP during pig adipocyte development. In addition, maturity of intramuscular and subcutaneous adipose tissue was studied by examining the expression of H-FABP, as well as, other adipose marker genes in both tissues. These studies suggest that H-FABP may play a role in adipose tissue development and function in the pig.<br /> <br /> 10. Experiment Station: University of Illinois; Project objectives: 1,2,6,7. Chronic Wasting Disease (CWD) is the only prion disease extant in free-ranging animals and is transmissible to livestock by experimental inoculation. A frequent symptom of prion disease in several species is loss of weight with either normal or reduces intake reflecting neural damage to body weight or feeding centers. To look for genetic resistance, nucleic acid sequences of the prion gene (Prnp) were examined and genotypes compiled for 182 white-tailed deer in northern Illinois, 62 of which were positive for CWD. Nine nucleotide polymorphisms, seven silent and two coding, were found in the sampled population. Five polymorphic loci demonstrated significantly different distributions of alleles between infected and non-infected individuals. Nucleotide base changes 60C/T, 285C/A, 286G/A, and 555C/T were observed with higher than expected frequencies in CWD negative animals suggesting disease resistant patterns, while 153C/T was observed more than was expected in positive animals, alluding to susceptibility. Both coding polymorphisms, Q/H95 and G/S96, have been previously described in white-tailed deer populations sampled in CO and WI. Despite similarities in observed genotypes, frequency distributions of coding polymorphisms in WI and IL deer populations were significantly different from each other, a surprising result considering the sampled areas are less than 150 km apart. The number of polymorphisms per animals, silent or expressed was highly correlated to disease status, with negative animals having more base changes than positive animals. An additive association of silent polymorphism number with disease resistance has not been previously reported in prion diseases. Most beef marketed in the US is aged to improve tenderness and consumer acceptability. Because of the inconsistencies in tenderness changes during aging, we attempted to look at the relationship between aging and initial tenderness irrespective of animal age, genetics or nutritional status. Cattle were selected to represent a full range of quality grades from Utility to Prime. Steaks from these cattle were aged in vacuum bags for 0 to14 days and tenderness assessed. Shear values decreased with aging time across most grade categories, however, improvement was not uniform. Shear values overlapped a great deal across all USDA grades so to gain a better understanding of changes with aging, steaks were grouped based on initial shear value differences of 1 kg. Shear value of the toughest steaks decreased the most during aging (-36%), while steaks with the lowest shear values actually toughened as shears increased (16%). The same pattern emerged when steaks were grouped by initial tenderness. These results suggest that initially tough steaks will benefit from aging while the quality of very tender steaks may decline with aging.<br /> <br /> <br /> Dietary Constituents Regulating Lipid Deposition or Lipid Composition:<br /> 1. Experiment Station: The Ohio State University; Project objectives: 8, 9, 10. Over-consumption of nutrients relative to energy expenditure is a main cause of weight gain, leading to obesity. The brain has important glucose-sensing neurons that detect fluctuation of blood glucose levels at fasting and feeding stages and responds by alteration in synthesis and secretion of neuropeptides to regulate food intake. The role of glucose transporter 2 (GLUT2), a possible glucose sensor in the brain in regulation of food intake in neuronal cell line, was investigated. Overexpression of GLUT2 gene increased ATP/AMP ratios, which inactivates AMP activated protein kinase, leading to suppression of the hunger signal in the neuronal cells. This finding on the function of GLUT2 in regulation of food intake will not only help us better understand the mechanism of glucosensing, but allow us to develop strategies targeting GLUT2 for regulation of food intake in obese subjects. <br /> <br /> 2. Experiment Station: The Ohio State University; Project objectives: 5. Most people on a Western diet consume much lower levels of n-3 FA than the recommended daily amounts and much higher levels of n-6 FA than is ideal for good health. Few of these people routinely eat the fish that are good sources of 20:5n-3 and 22:6n-3. Further, there is increasing concern about contaminants such as mercury and dioxins that may limit the healthiness of high fish intake. On average, Americans eat four times as much beef as fish, and studies have shown that cattle consuming forages or feeds containing n-3 FA can produce meat with higher levels of n-3 FA and lower levels of n-6 FA. We examined the ability of n-3 FA in flaxseed-supplemented rations to increase the n-3 FA content of bovine muscle. A significant increase in n-3 FA levels in bovine muscle from cattle fed rations supplemented with flaxseed and increased expression of genes that regulate lipid metabolism was observed.<br /> <br /> 3. Experiment Station: University of New Hampshire; Project objectives: 1, 2. The goal of this discovery-based project is to expand knowledge about the levels of polybrominated diphenyl ethers (PBDEs) in breast milk of lactating women from the Seacoast region of New Hampshire and to examine potential relationships between breast milk PBDE levels and maternal characteristics, their living environment and dietary intake. Women, ages 22 to 40 who were in the early stages of lactation (less than two weeks) or in the last trimester of pregnancy and planned to breastfeed for at least the first three months postpartum, were eligible for the study. Participants were asked for a maximum of nine breast milk samples during the first three months of breastfeeding. Each participant provided up to three samples at the end of their first, second and third month of breastfeeding for evaluation of day-to-day and month-to-month variation in PBDE levels. Participants were asked to complete four questionnaires during their 3-month commitment in the study, which provided maternal, diet and environment information. A total of forty women completed the study from November 2005 to October 2006 with an average age of 31 years. There was limited diversity among participants; 39 out of 40 participants were Caucasian, and 80% of participants had completed a 4-year college degree or higher. To date, 20 out of 40 breast milk samples have undergone PBDE analysis for eight major congeners. Participant means over the three month collection period was 7.69 to 139.78 ng/g lipid. Data from the first five participants revealed that variation from day to day was minimal with the exception of one outlier during month 2, day 2, so it was decided to only collect one sample per month from each participant. To date, month-to-month variation indicates a downward trend with monthly means at 41.52 ng/g lipid for month one, 38.26 ng/g lipid for month two and 36.40 ng/g lipid for month three. The next steps in the project include awaiting the remainder of the PBDE data, which will allow for an examination of potential relationships between PBDE levels and maternal characteristics, living environment and dietary intake. <br /> <br /> 4. a. Experiment Station: University of Georgia; Project objectives: 1, 5, 8. Effects of dietary fish oil in gestation or lactation on performance of progeny were investigated. Fish oil was fed as a protected product (Fertilium) in the first experiment beginning at D 60 of gestation through weaning. The study was conducted as a 2 x 2 design with fish oil fed in gestation, lactation or both. Milk was collected at days 1, 7, and 21. Pig performance was monitored through market weight. Selected pigs were challenged with LPS at 14 days of age. The effect of maternal diet on cytokine and cortisol response to an LPS challenge was determined. The addition of the product to the maternal diet resulted a lower n-6/3 ratio (Control 18-20, fish oil 10-12). There was no effect of dietary fish oil on growth performance or ultrasound carcass composition. Despite the low inclusion rate(less than 0.5% EPA and DHA) in the diet, milk fatty acid profile was altered in sows fed fish oil. Pigs from sows fed fish oil had an attenuated cortisol and TNF-± response to LPS challenge.<br /> <br /> b. Experiment Station: University of Georgia; Project objectives: 1, 2. In the second experiment, sows were fed diets containing 2 % soy oil or 2% fish oil during lactation. Data was collected as in the previous study and is being analyzed. A third study was conducted to determine the effect of maternal fish oil consumption on the fatty acid profile of pigs at birth and day 14. Sows were fed 0.5% fish or soy oil beginning at either day 60 or farrowing. Pigs were obtained at birth and day 14. Fatty acid profile of brain, adipose tissue, muscle, liver, lung, heart and kidney will be determined.<br /> <br /> <br /> c. Experiment Station: University of Georgia; Project objectives: 1, 2. This study was conducted as a 2 x 2 factorial with main effects of consuming an omega-3 supplemented diet during growing and/or finishing. Pigs (n=80, barrows and gilts) weighing approximately 25-30 kg were allotted by gender and weight to either Control or Fish Oil diet treatments. The Grower diets were corn-SBM based diets calculated to contain approximately 18% CP, 1% Lysine, and 3327 kcal/kg ME . At d 35 of the study half of the pens in each diet treatment were maintained on the same diet and half switched to the other diet (Control or Fish Oil) for finishing. The Finisher diets will also be corn-SBM based diets calculated to contain approximately 17% CP, 0.9% Lysine, and 3340 kcal/kg ME. At d 70 of the study all pens were switched to a common finisher diet calculated to contain approximately 16% CP, 0.8% Lysine, and 3345 kcal/kg ME. The study ended at d 98. Pig weight and feed intake per pen was recorded weekly. Ultrasound images of the 10th rib loin area was recorded at d 70 (when switched to common diet) and at the end of the study (d 98) in order to evaluate growth parameters and days to finish. Representative pigs were processed at week 5 and 10 to determine th immediate effects of diet on fatty acid profile. One male and one female from each pen was slaughtered at the end of the study period to obtain tissue samples and carcass data for the purpose of determining tissue fatty acid composition and any possible effects on the meat of long term fish oil supplementation in the diet. <br /> <br /> d. Experiment Station: University of Georgia; Project objectives: 1,2. In previous studies we have shown that composition of gain (protein and lipid accretion rates) influence energy and protein intake. In somatotropin treated pigs, lipid accretion is reduced and protein accretion is stimulated. Pigs allowed to self-select between high and low crude protein diets alter their intake patterns to match the changes in composition of gain. They consume less of the low CP diet in response to reduced caloric needs and tend to increase consumption of the high CP diet in response to the greater rates of lean growth. Similarly, pigs with less backfat select a higher percent protein diet than pigs with greater backfat. The objective of this experiment is to determine the influence of Paylean on diet selection. Paylean increases loin area and reduces fat thickness and would be expected to have effects on selection that are similar to somatotropin. Growing barrows will be allowed to establish a selection pattern and then have Paylean treatment imposed. Twenty barrows with an initial weight of 80 kg were fed the UGA grower diet for 7 days to acclimate to the pens. On day 7 pigs were allowed to select from the high and low protein diets (without Paylean). Each pen had 2 feeders and the position of the high and low CP diets in the pen was changed daily. On day 21, half of the pigs received high and low protein diets supplemented with Paylean (20 ppm) and remained on these diets for an additional 2 weeks. Feed intake will be determined daily. Pigs will be weighed and ultrasounded at weekly intervals. During the pre-treatment period pigs self-selected 18-20% crude protein diets. During the treatment period, there was no difference in total intake between the groups. However, pigs supplemented with Paylean selected a higher percent protein than the control group. <br /> <br />

Publications

Impact Statements

1. 1. A method to study dedifferentiation of mature fat cells is being developed, that will apply to meat animal growth and development, as well as human obesity and diabetes.
2. 2. In adipose tissue of prepubertal gilts, short term feed restriction decreased lipogenic enzymes and transcription factors, while other important lipogenic enzymes and proteins were minimally affected
3. 3. The newly discovered adipocyte gene, Tusc5, has markedly reduced mRNA expression in white adipose tissue of obese Pima Indians, with smaller reductions observed in a French cohort of obese women, indicating the genes involvement in human obesity.
4. 4. In lactating beef cow adipose tissue, abundance of lipogenic enzyme mRNA in adipose tissue is influenced by the level of adiposity of the cows at parturition, and time in milk influences lipogenic and lipolytic enzyme mRNA in beef cows.
5. 5. A link between obesity and atopic airway disease was discovered in women; weight gain is related to development of eicosanoids that favor a type 2 immune response.
6. 6. Activation of PPARg2 by selective agonists for this transcription regulator could selectively alter adipogenesis in intramuscular adipose tissue of beef cattle.
7. 7. Single nucleotide polymorphisms for bovine fatty acid synthase could be used as a DNA marker to select breeding stock to alter the fatty acid profile of the meat to reflect increased healthfulness of the fatty acids in beef.
8. 8. Hepatic fatty acid binding protein plays a role in adipose tissue development and function in the pig.
9. 9. Over expression of the glucose transporter gene, GLUT2, plays a role in food intake regulation by increasing ATP/AMP ratios, which inactivates AMP activated protein kinase, leading to suppression of the hunger signal in neuronal cells.
10. 10. Polybrominated diphenyl ethers appear in lipids of human breast milk at levels that tend to decrease with time in lactation, and levels of these compounds likely vary with maternal characteristics, living environment and dietary intake.
11. 11. Pigs consuming milk from sows fed 0.5% EPA and DHA had attenuated cortisol and TNF-a response to LPS challenge.
12. In addition, NCCC097 continues to encourage and engage scientists in productive collaborations. The most important collaborations that arise from NCCC097 are the interactions at the annual meeting. Here we discuss and share methods, float new ideas and debate widely diverging topics within lipid and adipose tissue metabolism. While almost impossible to document, the "workshop" type atmosphere in our meeting allows all to benefit greatly.
13. Over the past few years, ongoing collaborations have been fruitful. Dr. Werner Bergen worked cooperatively with Dr. Harry Mersmann and Dr. Jack Odle, and a paper was published on "Comparative Lipid metabolism" in Journal of Nutrition in 2005. Cooperative work between Drs. Bergen and Odle resulted in a joint USDA-NRI proposal (not funded).
14. Dr. Steve Smith has collaborated with Dr. Hiroyuki Kawachi and Dr. Hideo Yano, Kyoto University, Japan; with Dr. Chang Bon Choi, Yeungnam University, Korea; Dr. Chang Weon Choi, National Livestock Research Institute, Suwon, Korea; Dr. Ron Tume, CSIRO, Brisbane, Australia; and with Dr. Guoyao Wu, Texas A&M University. These collaborations are focused on the effects of arginine on adiposity in livestock species.
15. Drs. Carey and Rule collaborated on a project wherein fat from breast milk obtained from women who consumed a conjugated linoleic supplement was analyzed for fatty acid concentrations. A paper is currently being prepared for submission for publication.
16. During the present reporting period, collaborations occurred between the Washington State University Station and the USDA, ARS Station in Georgia on a research project intended to define if mature adipocytes possess the ability to dedifferentiate into proliferative-competent progeny cells. Conditions were defined in which lipid-filled adipocytes were shown to divide, in vitro, and produce progeny adipofibroblasts. Progeny adipofibroblasts, however, displayed a protracted ability to redifferentiate into lipid-assimilating adipocytes, in vitro. Findings resulted in 3 published papers.

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Report Information

Participants

Adams, Sean (sean.h.adams@ars.usda.gov) - USDA Western Human Nutrition Research Center;
Ajuwon, Kolapo (kajuwon@purdue.edu) - Purdue University
Azain, Michael (mazain@uga.edu) - University of Georgia;
Barnes Kimberly (KMBarnes@mail.wvu.edu) - West Virginia University;
Beitz, Donald (dcbeitz@iastate.edu) - Iowa State University;
Bergen, Werner (bergewg@auburn.edu) - Auburn University;
Dunn, Tamara (tamara.n.dunn@ars.usda.gov) USDA Western Human Nutrition Research Center;
Hausman, Gary (ghausman@saa.ars.usda.gov) - USDA, Athens, GA;
Kelly, Amy (amykelly@illinois.edu) - University of Illinois;
Kinder, James (kinder.15@osu.edu) - Administrator for Project, The Ohio State University;
Knotts, Trina (trina.a.knotts @ars.usda.gov) - USDA Western Human Nutrition Research Center;
Lee, Kichoon (lee.2626@osu.edu) - The Ohio State University;
Mark Mirando (mmirando@csrees.usda.gov) - Administrator, USDA-CSREES;
Nafikov, Rafael (rnafikov@iastate.edu) - Iowa State University;
Novakofski, Jan (jnova@illinois.edu) - University of Illinois;
Odle, Jack (jack_odle@ncsu.edu) - North Carolina State University;
Rule, Daniel (dcrule@uwyo.edu) - University of Wyoming;
Sainz, Roberto (rdsainz@ucdavis.edu) - University of California-Davis
Schoonmaker, Jon (jschoon@iastate.edu)- Iowa State University;
Xi, Lin (lin_xi@ncsu.edu) - North Carolina State University;
Yu, Xing (xingyu@isisph.com) - Industry guest, Isis Pharmaceuticals

Brief Summary of Minutes

Summary of minutes of annual meeting:

Dr. Rule opened the meeting by introducing Drs. Kinder and Mirando, then new members, graduate students and visitors. Dr. Hausman and Dr. Novakofski were elected as co-chairs for the 2009 meeting. The committee decided to continue meeting in conjunction with Experimental Biology, which will be in New Orleans for 2009. The committee discussed hosting a symposium at EB in 2010 or at ASAS in 2009. Additional outreach in the form of a JAS review article with Hausman as lead author was discussed. The issue of attendance was discussed and the committee agreed to send an e-mail before the meeting stressing the importance of attendance, and to revisit the issue of members that were not attending at the 2009 business meeting. Non-attending members with no other evidence of interest or commitment would be administratively removed in the future.

Dr. Kinder gave a brief administrative update, and stressed the importance of communicating the value of the group activities, especially impacts and output, in annual reports and summaries. Dr. Kinder reminded the committee that the project renewal was due before December 1, 2008 and stressed the involvement of pos-docs and grad students and the balance between research, teaching and outreach activities.

Dr. Mirando discussed potential factors that might impact funding opportunities for NCCC097 members, as well as current and projected CSREES budgets. Dr. Mirando also discussed the potential to use outreach activities as research.

Project Directors, graduate students, and postdoctoral researchers provided research update (see Accomplishments) followed by discussion and questions for each project. Thirteen research updates were presented by committee members with a guest presentation on from Xing Yu, Isis Pharmaceuticals, who had attended NCCC097 meetings as a student.

Accomplishments

Accomplishments:<br /> <br /> The research objectives for NCCC097, October 2004 to September 2009, are available at the NCRA website: (http://lgu.umd.edu/lgu_v2/homepages/outline.cfm?trackID=3848) 1. Share and critique new techniques, experimental designs, and new unpublished data regarding adipocyte biology. 2. Through the interactive process at the meeting and afterward, elicit input to improve experimental design and to develop mutual interests toward joint projects. 3. Study biological regulation of catabolic (fatty acid mobilization and oxidation) and anabolic (fatty acid synthesis, elongation and desaturation, and triacylglycerol synthesis) lipid metabolism in domestic species at the molecular, cellular, tissue, and organismal level 4. Study cellular and molecular controls of adipocyte hyperplasia, differentiation and growth in domestic species. 5. Study nutrient determinants of excess fat deposition (diet composition, and amount and source of dietary fat, calories, and protein). 6. Study mechanisms associated with genetic variation in excess fat deposition and the post-genomic variation of individual phenotypes (determinants of cellularity, receptor populations, alternative metabolic signalling pathways, alternative gene expression and promoter function) 7. Establish approaches to maintain or enhance intramuscular fat concentration, but at the same time maintain or decrease subcutaneous and intermuscular fat. Intramuscular fat (marbling) is an important contributor to the organoleptic properties of meat, but the molecular and cellular mechanisms that retard intramuscular fat deposition until late in development are not understood. 8. Investigate the interplay between adipose tissue, liver, gut, and skeletal muscle lipid metabolism. The growth of adipose tissue is regulated, not only within the adipocyte, but also by the the interorgan interchange of metabolites, various lipoproteins, and numerous hormones and signalling molecules. 9. Investigate the role of endocrine factors produced by the adipocyte in co-ordinate regulation of feed intake, reproductive competency, energy expenditure, and immune function. 10. Study mechanisms for intervention strategies to decrease excess fat deposition (exercise, caloric restriction, pharmacotherapy, selective control of gene expression through transcription factors and modulation of promoter function). NCCC097 scientists conducted the following work over the past year. The Experiment Station and project member, followed by the objective number(s) met by the work, are provided for each entry. <br /> <br /> <br /> 1. Experiment Station: West Virginia. Representative: Kimberly M. Barnes. Objectives 1, 2 and 10. This year has been spent primarily setting up our laboratory and acquiring instruments. We have two primary projects planned for initiation this summer: The effect of oil source on conjugated linoleic acid-induced lipolysis in mice and The effect of feeding high-DHA Crypthecodinium cohnii on body composition in mice. Previously, we demonstrated that mice fed coconut oil from weaning for 6 weeks lost more body fat when supplemented with CLA than did mice fed soy oil. Part of this greater response appears to be due to increased basal lipolysis observed in CLA-fed mice raised on coconut oil but not soy oil. In cell culture models increased lipolysis in response to CLA has been observed and associated with increased activation of perilipin. This is followed by de-differentiation of the adipocytes, marked by decreased perilipin expression and increased adipose differentiation-related protein (ADRP). We plan to measure the phosphorylation (indicating activation) and expression of perilipin and hormone sensitive lipase and the expression of ADRP in reteroperitoneal fat pads of mice fed soy oil, soy oil + CLA, coconut oil, or coconut oil + CLA. Further studies on the activation of signaling molecules and a time-course of lipolysis activation are also planned. A colleague at West Virginia University, Dr. Kristen Matak, has developed a continuous culture system to grow Crypthecodinium cohnii. This algae has the potential to accumulate a large percentage of its biomass as lipid and to be highly enriched for the omega-3 fatty acid DHA. In Dr. Mataks system the algae has been determined to be over 18% lipid, with almost 30% of that lipid as DHA. We have determined that fish oil has the potential to lower body fat percentage alone but to inhibit CLA-induced body fat loss. <br /> <br /> 2. Experiment Station: North Carolina State University. Representative: Jack Odle. Objectives 1, 2, 3 and 8. Infant formula companies began fortifying formulas with long-chain polyunsaturated fatty acids (PUFA) in 2002. Long-chain PUFAs are precursors for the synthesis of prostanoids which stimulate recovery of intestinal barrier function. The objective of these experiments was to investigate the effect of higher supplementation of AA on intestinal barrier repair in ischemic-injured porcine ileum. One day-old pigs (n=24) were fed a milk-based formula for 10 d. Diets contained either no PUFA (0%AA), 0.5%AA, 5%AA, or 5% eicosapentaenoic acid (EPA) of total fatty acids. Following dietary enrichment, ileum from piglets were subjected to in vivo ischemia injury by clamping the local mesenteric blood supply for 45 min. Following the ischemic period, control (non-ischemic) and ischemic loops were harvested and transepithelial electrical resistance (TER) was measured over a 240 min recovery period. Ischemia-injured ileal tissues from piglets fed the 5% AA diet and 5% EPA exhibited enhanced recovery (% increase in TER = 13±13, 21.6±12 59.1±12, 50.8±13, for 0%AA, 0.5%AA, 5%AA, and 5%EPA, respectively, p<0.05). Enhanced TER recovery response observed with 5% AA supplementation was attenuated by in vitro application of indomethacin suggesting responses were mediated by prostaglandins. Histological evaluation of piglets fed the 5% AA formula showed reduced lesions after ischemia compared to other dietary treatments (P<0.05). Long-chain PUFAs enhances recovery of TER in ischemic injured porcine ileum mediated via endogenous prostaglandins.<br /> Objectives 1, 2, 3, 5 and 8. Another study examined the effect of maternal feeding of 0.8% clofibrate for 7 days on fatty acid oxidation in newborn pigs. The maternal-fed clofibric acid had no impact on the liver weight of the newborn pigs, but hepatic fatty acid oxidation examined in fresh homogenates showed that clofibrate significantly increased 14C-accumulation in CO2 and ASP by 2.9-fold from [1-14C]-oleic acid and 1.6-fold from [1-14C]-lignoceric acid. Correspondingly, hepatic carnitine palmitoyltransferase (CPT) and fatty acid oxidase (FAO) activities were increased also by 36% and 42% compared to the newborn pigs from the control sow. Clofibrate crosses the placental membrane of the sow and enters fetal circulation. Thirty four colostrum-deprived newborn pigs were fed milk replacer and orally gavaged with either vehicle (2 % Tween 80) or clofibrate (75 mg /kg body weight) +/- etomoxir (5 mg/ kg body weight) once daily for up to 7 days, and the sensitivity of liver CPT I to malonyl-CoA inhibition and fatty acid oxidation were determined using 1-14C-octanoic, oleic and erucic acids. Administration of clofibrate significantly increased liver CPT I activity (60%) and malonyl-CoA IC50 values (56%). Addition of etomoxir potently inhibited CPT I activity, but malonyl-CoA IC50 remained unchanged. Consistent with CPT I activity, clofibrate increased mitochondrial fatty acid oxidation by 53%, but did not affect the rate of peroxisomal beta-oxidation. Etomoxir did not alter the rate of fatty acid oxidation despite a 30% reduction in CPT I activity. Fatty acid oxidation induced by clofibrate is due to both an increase in CPT I activity and a decrease in sensitivity of CPT I to malonyl-CoA inhibition.<br /> Objectives 1, 2, 8 and 9. Another research area was safety assessments of novel infant formula ingredients in neonatal pig models. One-day old pigs were fed a cows milk-based formula supplemented with PDX (0, 2, 5, 10 or 20 g/L) for 18 days (n=6/formula group) to further substantiate the efficacy and safety of the ingredient. Additional reference groups included 6 pigs sampled at day 0 and 12 sow-reared pigs. Growth rate, formula intake, stool consistency, behavior score, blood metabolites and relative organ weights (i.e. liver, spleen and kidney as % of body weight) did not differ among formula-fed pigs (P > 0.1). Among PDX-fed pigs, digesta pH decreased linearly (P < 0.05; ileum, cecum, colon) and quadratically (P < 0.05; ileum, cecum) as dietary PDX increased, with a maximal reduction approaching 0.5 pH units in those fed 20 g/L. Cecal and colonic pH of sow-reared pigs were similar to pigs fed 20 g/L PDX. Polydextrose shows promise as a prebiotic additive in infant formula.<br /> <br /> 3. Experiment Station: Washington Station. Representative: M.V. Dodson. Objectives 1, 3, 4 and 7. Cell culture model of adipocytes has proven to be both a boon and a bust to scientists. If the cell culture system is well developed, and time is not required to establish the limits of the in vitro system, then relatively large amounts of research data may be generated in relatively short timeframes and researchers may make steady research gains and publish above average numbers of papers with minimal delays. However, cell culture systems and conditions differ between laboratories. In many cases data generated under similar conditions but in different laboratories are not consistent. For adipogenesis studies, cell lines and stromal vascular cells are commonly used, even though the value of each system is limited. Between-laboratory comparisons may NOT occur if the cultures are not maintained similarly/properly, are/become transformed, or if the entire cellular system is ill-defined (cells such as SV cell cultures can NEVER be enumerated for consistent subpopulation components between laboratories). Vigilance is required to obtain repeatable data from these systems, especially if mechanistic or developmental experimental designs are attempted. We suggest that all aspects of basic cell culture are as important as growing cells. Our progeny cell cultures are not like SV cells or cell lines in dynamics of re-differentiation and formation of lipid-filled adipocytes.<br /> Objectives 1, 3, 4 and 6. Another project examined gene regulation of bovine-derived adipofibroblasts. It is important to understand mechanisms of fat metabolism in beef cattle as the fat component and distribution are directly associated with the quality and the value of the meat. The discovery of gene markers associated with various carcass traits in beef cattle has created links between the genetic background of the animal and its performance. Animal performance is directly associated with the end products of gene expression and the functional proteins expressed. Beef-derived adipocytes (derived from mature adipocyte dedifferentiation in Dr. Dodson's laboratory) were analyzed by Dr. Leluo Guan (University of Alberta), with genomic/proteomic tools. Using perimuscular-derived adipocyte-derived preadipocytes, Dr. Guan showed that no C/EBP± was expressed by differentiating beef-derived cells in vitro. As C/EBP± operates with other nuclear signaling agents (especially PPAR³), either PPAR³ functions alone in regulating adipogenesis in the perimuscular adipose tissue depot of beef steers, or the expression of C/EBP± is transitory. Adipocytes from one adipose tissue depot in beef cattle are different to other types of fat cell systems, or other adipose depots.<br /> Objectives 1, 3 and 7. Adipose tissue contains a large portion of stem cells and it is important toidentify transdifferentiated cells from progeny cultures derived from dedifferentiation of mature adipocytes in vitro. These cells appear morphologically like fibroblasts, and are primarily derived from the stromal cell fraction. Mature (lipid-filled) adipocytes possess the ability to become proliferative cells and have been shown to produce progeny cells that possess the same morphological (fibroblast-like) appearance as the stem cells from the stromal fraction. This simple fact may prove to provide a novel mechanism via which we can learn about the stem-cell nature of the progeny cells, determine if the proliferative-competent progeny cells are capable of becoming other cell types, and capitalize on the properties of the progeny cells for tissue engineering purposes. <br /> Objective 1 and 2. Many dynamics of domestic animal physiology, adipogenesis and assimilation of energy are different from rodents and/or humans. We periodically submit scientific papers to help teach scientists in other disciplines about domestic animal-derived adipocyte research. Historical research with adipose tissue/adipocytes proved that tissue/cells actually played a large role in the outcome of any given experiment. Much of today's research with adipose tissue/cells, however, is focused on subcomponents of the cell (DNA/RNA), intracellular regulatory pathway (signaling mechanism) of cell morphogenesis, or the possible identification of adipose tissue-derived stem cells for tissue engineering purposes. Years ago, both tissue characteristics and the source of the cell were quite important. Today, what is important is the molecular mechanism being defined. Considering the molecular emphasis placed on graduate training in this field, it is easy to suggest that new investigators might be incapable of extrapolating their molecular data to the cell, tissue or live animal in any practical manner. One goal of my work is to provide historical reference to those interested in adipose tissue/cells in animal sciences. <br /> <br /> 4. Experiment Station: Western Human Nutrition Research Center. Representative: Sean H. Adams. Objectives 1, 2, 4, 5 and 6. We introduced our initial characterization of tumor suppressor candidate 5 (Tusc5) at the 2006 and 2007 NCCC097 meetings, and published on this gene in 2007. We have extended those findings by identifying and characterizing a second gene-protein with unique robust expression in both afferent neurons and adipocytes: g-synuclein (SNCG). Searching databases for expression similar to Tusc5, and follow up by RT-PCR, confirmed high expression in these sites in humans, mice, and rats. This protein has been previously implicated in cancer tissue and its knockdown limits cancer cell line proliferation. With James Pan, Toronto we found that the gene is downregulated by PPARg agonists and is induced during 3T3-L1 differentiation. Our model is that SNCG is involved with fat tissue (and neuron) plasticity in response to metabolic cues. SNCG mRNA expression is significantly increased in the WAT of obese humans and is highly expressed in subcutaneous vs. visceral/omental fat (confirmed in several human cohorts). Its expression patterns are closely correlated with those of leptin in both cell culture and human WAT, indicating shared regulatory elements. Since SNCG was reported to physically interact with TIMP-2, a regulator of extracellular matrix remodeling, future studies will examine how SNCG influences the adipose ECM in response to obesity and PPAR modulation. <br /> Objectives 1, 2, 6, 9 and 10. Low muscle fatty acid catabolism is associated with insulin resistance. Identification of biomarkers of muscle fat combustion could prove useful to predict disease or monitor efficacy of interventions. No metabolite or metabolite pattern has specifically been tracked to muscle fat catabolism. Comprehensive acylcarnitine analyses and clinical chemistry were conducted to compare plasma of controls vs. persons harboring a missense mutation in UCP3 (a muscle-specific protein in humans; the missense yields 50% lower fat metabolism and a higher CHO catabolism), in collaboration with Drs. Tim Garvey (UAB) and Chuck Hoppel (Case Western). Few differences in metabolites were observed when comparing wildtype vs. UCP3 polymorphic subjects, with the exception of butyrylcarnitine and lactate that were lower in the latter. Comparing diabetics vs. non-diabetics across genotypes, diabetics displayed significantly increased acylcarnitine:free carnitine ratio, higher acetylcarnitine, and reduced propionylcarnitine. Current results are consistent with the hypotheses that: (a) persons with dysfunctional UCP3 in muscle improve efficiency of pyruvate flux toward the TCA cycle (hence, lower lactates), (b) C4-carnitine may be a sensitive marker of ß-oxidation/CHO oxidation dynamics, and (c) type 2 diabetics may have more limited TCA cycle function reflected in higher acetylcarnitine levels in plasma resulting from reduced capacity to dispose of acetyl-CoA. Lower C3-carnitine in diabetics might mark a more limited generation of propionyl-CoA, an important anaplerotic carbon source for replenishment of the TCA, which if true could contribute to compromised TCA cycle function. <br /> <br /> 5. Experiment Station: University of Geogia. Representative: M. J. Azain. Objectives 1, 2, 7 and 10. The 14 wk study was conducted as a 2 x 2 factorial with main effects of consuming an omega-3 supplemented diet during growing and/or early finishing phase. The n-6/n-3 ratios of the control and fish oil diets were in the range of 10-12 and 2-3, for the soy and fish oil diets, respectively. There was no effect of dietary fat source on growth performance. There were only minimal effects of diet on carcass characteristics. There was a trend for greater lipid content in the loin muscle from pigs fed fish oil for the first 5 weeks of the study. At slaughter, average lipid % was 4.23% in pigs fed the control diet for the first 5 weeks as compared to 5.04% in pigs fed fish oil (P < 0.10). As expected, dietary fat affected tissue fatty acid profiles. The n-6/n-3 ratio in loin muscle of control pigs was 18.7, 17.7 and 20.9 in tissues sampled on days 35, 70 and 98. In contrast, the ratio was 4.4, 3.7 and 5.8 on days 35, 70 and 98, respectively, in pigs fed fish oil from day 0-70. Feeding fish oil for only the first or second 5 week periods resulted in intermediate ratios. Calculated long-chain omega-3 content was increased from less than 5 mg per serving of pork to 30-60 mg/serving in pigs fed fish oil for 5 to10 weeks. Although a taste panel test was not conducted, there were no obvious fishy odors in any of the samples on day 98 (4-9 week withdrawal period). The increase in intramuscular fat and changes in fatty acid profiles in response to feeding fish oil in the grower period, are worth of further investigation.<br /> Objectives 1, 2, 3 and 7. A significant portion of the US corn crop is currently going into ethanol production. This has resulted in greater feed costs, causing producers to examine alternative feed ingredients. Distillers dried grains with solubles (DDGS) is a co-product of ethanol production from corn and has received considerable attention with expansion of ethanol plants. DDGS is essentially corn without the starch and thus, is enriched in protein, ether extract and fiber. Fermentation converts much of the phytin phosphorous in corn to a more readily available forms. One issue with use of DDGS in swine diets is the higher level of ether extract in the product. Since this is essentially corn oil, it is an unsaturated fat source and feeding may result in higher iodine values which reduces the acceptability of pork by the packing industry and reduces shelf-life in the display case. A means to offset the negative effect of DDGS on pork carcasses would be advantageous. We determined if addition of a saturated fat to the diet counteracts the negative effects of the unsaturated fat in DDGS on pork quality. The study was conducted as a replicated 2 x 2 design with main effects of saturated or hydrogenated fat addition (0 and 4% Fat Pak 100, Milk Specialties), and DDGS ( 0 vs 30% ) inclusion. Pigs fed diets with 30% DDGS had reduced intake during the first 2 weeks of the study and overall. Intake was not different from week 2-6, but pigs did not recover from the reduction in the first 2 weeks. As a result, overall growth rate and final body weight was reduced in pigs fed DDGS. Tissue fatty acid profiles were altered by DDGS feeding. Calculated iodine value in subcutaneous, belly and leaf fat, but not loin muscle, was increased with DDGS. This was associated with a greater linoleic acid content in tissues from pigs fed DDGS. The addition of hydrogenated fat product, Fat Pak, had minimal effects on fatty acid profiles. The lack of effect of this fat source on tissue profiles is accounted for by the lack of digestibility of the product. <br /> <br /> 6. Experiment Station: University of California Davis. Representative: Dr. Roberto D. Sainz. Objectives 1, 2, 4, 5, 6 and 7. The beef and other meat industries are becoming more competitive and more uncertain, due to environmental concerns, increases in commodity prices and diversity in feed availability for animals. Prices received by producers are based upon the carcass value of their animals (quality and yield grades for beef cattle). The primary determinants of carcass value are the amount and distribution of carcass fat. The key to maximizing carcass value and profit is to feed animals to their optimal endpoint with adequate marbling and not excessive fat trim. Thus there is a need for tools to guide producers in their management decisions. Mathematical models of animal growth and body composition have been shown to accurately predict performance and composition of animals of different genotypes under different management scenarios. To predict the optimal slaughter endpoint, a model must be dynamic, i.e., the model should predict gain and composition over a period of time using known relationships between state variables. The best models currently available are also deterministic (there are no random elements) and mechanistic, which means that equation forms and parameter values are based on knowledge of the underlying biology. Current models (e.g., NRC, 2000) are static and empirical (relationships are based on regressions), so cannot be applied to production scenarios beyond the range of values used in their development. The Davis Growth Model (DGM) is a dynamic, deterministic and mechanistic model of beef cattle growth and body composition (Oltjen et. al., 1986; Sainz and Hasting, 2000). The DGM is based on fundamental biological concepts of tissue growth, such as hyperplasia and hypertrophy, as well as net energy. Body protein accretion is calculated based on the difference between the rates of protein synthesis (determined by total DNA content, hormonal status and plane of nutrition) and protein degradation. Accumulation of total body fat is determined from the net energy available after accounting for maintenance and protein gain. Body fat is further partitioned among four depots. The DGM predicts overall growth and body composition of cattle of different genotypes, implant status and feed regimes with excellent accuracy and precision. The equations to partition fat gain among of adipose depots, however, was based upon only two data sets (Robelin, 1981; Cianzio et al., 1985), as these were the only publications found in which fat depot weights, cell numbers and sizes were reported. As a result, the DGM can be used to accurately forecast body composition over time, but predictions of fat distribution are inadequate. To improve these equations, more data on adipose cellularity in animals of different genotypes, ages, and growth trajectories will be required. <br /> Objectives 1 and 2. The objective of this research is to investigate the effect of breed, sex, implant status and energy level of the diet on adipocyte development and distribution in beef cattle. Animals will be harvested at different ages and fat depot weights will be determined by dissection. Samples from each the four fat depots will be collected, and prepared for histological examination of cell numbers and sizes. Another goal is to develop a faster method to make those measurements in the cold carcass, in order to enable data acquisition on a larger scale in the future. In addition to measurements of adipocyte cellularity, transcriptional factors involved in regulation of adipogenesis will be determined. These will include C/EBP, PPAR³, and ADD1/SREBP. Other factors may include adiponectin, leptin, and other adipocyte-specific markers. The exact nature of the molecular analyses will depend on consultations and collaborations with other members of the committee. The relationships and equations will be specific to beef cattle, but the underlying biology should be applicable to other species including humans.<br /> <br /> 7. Experiment Station: Auburn University/ Alabama Agricultural Experiment Station. Representative: Werner G. Bergen. Objectives 1, 2, 3,4, and 6. We haveshown that Duroc and Pietrain F1 pigs differ in lipid deposition and fatty acid oxidation in liver, adipose and skeletal muscle. Finishing Duroc pigs had a greater propensity for de novo fatty acid synthesis and a lesser propensity for fatty acid oxidation than finishing Pietrain pigs based on mRNA abundance for key lipogenic and beta oxidation enzymes. We have continued to explore metabolic differences in fat vs. lean type pigs by assessing differential expression of PGC-1± and UCP-3 in skeletal muscle. The expression results below are based on qRT-PCR assays. For both PGC-1± and UCP-3 there are no apparent differences in mRNA abundance in control pigs; however the beta adrenergic agonist marginally enhanced UCP-3 expression in both pig genotypes while was PCG-1± slightly enhanced in both breeds. We are presently also evaluating the relationship between residual feed efficiency (RFI), feed efficiency and expression of trans- acting factors, coactivators and lipid anabolic and oxidative genes in skeletal muscle and adipose tissue of beef cattle. Tissue samples were secured during the last week of a finishing trial by biopsy and gene expression was assessed using qRT-PCR. There were 8 lower RFI bulls and 8 higher RFI steers (Angus sired from Angus X Simmental dams). The RFI values are not yet available; however the bulls had a lower F/G (5.82) than the steers (7.20). The mRNA abundance in adipose biopsies for PGC-1± were not different between the bulls and steers. <br /> <br /> 8. Experiment Station: University of New Hampshire. Representative: Gale B. Carey. Objectives 1, 4, 6, 8, and 9 Our recent publication (Obesity 15:2942-2950, 2007) describes the effect of administering polybrominated diphenyl ethers (PBDEs), a family of flame retardants, to rats daily for 4 weeks, on isolated adipocyte metabolism. These lipid-soluble, flame-retarding environmental chemicals will deposit in adipose tissue, and our findings demonstrate that adipocytes isolated from treated rats are 30% more sensitive to epinephrine and 60% less sensitive to insulin, compared to adipocytes from control rats. This adipocyte metabolic obesity occurs despite similar body weight and adipocyte size between treated and control rats. Our next step was to investigate two possible sites of insulin hormone disruption due to PBDE treatment of rats: insulin-stimulated glucose transport and GLUT4 levels, and insulin-stimulated glycolytic and TCA-cycle flux, in adipocytes from PBDE-treated vs. control rats. To our surprise  but, as weve since learned  not uncommon to adipocyte researchers - our first few experiments revealed that the insulin-stimulation of glucose oxidation (measured by 14C-glucose conversion to 14CO2 over a 90-minute incubation), in adipocytes from control rats (~350 gms), had vanished. Over the next eight months, experiments were conducted testing different amounts and sources of albumin, collagenase, adenosine, new plastic ware, all new reagents, a different source of water, different ages of rats, even conducting the experiment in a different lab. Thirty experiments later, there was still no reproducible insulin effect. One variable, identified in literature published ~40 years ago, that we did not control was the timing and amount of food consumed. Therefore, we conducted a meal-feeding experiment, in which rats were adapted to eating their food in 2 hours/day. We measured insulin-stimulated glucose oxidation in adipocytes from 180 grams rats immediately after eating (n=2), 5 hours later (n=2) and 10 hours later (n=2). Weve had 6 successful experiments with a 5-fold stimulation of glucose oxidation in response to insulin at each time point. <br /> <br /> 9. Experiment Station: Iowa State University. Representative: Donald Beitz. Objectives 1, 2, 3, 5, 6, and 9. Elevated blood cholesterol and low-density lipoprotein-cholesterol are the main factors contributing to the CVD development in humans. The concentration of those metabolites in blood is increased when foods rich in saturated fatty acids are consumed. The major sources of those fatty acids are animal products that provide collectively 56% of total lipids, 74% of saturated fatty acids, and 100% of cholesterol consumed by humans. Decreasing the percentage of saturated fatty acids in milk and other animal products can lead to the improvement of the healthfulness of those foods and to decrease incidence of CVD in humans. Contributions of individual fatty acids to atherogenic potential for a lipid source or a diet are summarized by an atherogenic index (AI) that was developed by Ulbricht and Southgate in 1991. The AI is calculated as a ratio of the sum of concentrations of lauric (12:0), four times myristic (14:0), and palmitic (16:0) acids to the sum of concentrations of mono- and polyunsaturated fatty acids. The objective of this study was to determine if variations in single nucleotide polymorphism (SNP) in thioesterase domain of the fatty acid synthase (g.17924 A>G Threonine>Alanine) and diacylglycerol acyltransferase-1 (g.10433/10434 GC/AA Alanine>Lysine) genes would explain variations in milk fatty acid composition among Holstein dairy cattle. About 200 cows participated in the study. Milk samples were collected monthly throughout the first ten months of lactation and analyzed for milk fatty acid composition by gas chromatography. Blood samples were used to obtain a DNA sample for each animal. Cows with g.17924GG genotype had lower AI compared with cows of g.17924AG genotype (P=0.007). Likewise, cows with g.10433/10434GC genotype had lower AI compared with cows of g.10433/10434AA genotype (P=0.016). The decrease in AI for cows with g.17924GG and g.10433/10434GC genotypes was achieved by the decrease in the concentration of palmitic acid (P=0.06 and P<0.0001, respectively) and by the increase in the concentration of mono- and polyunsaturated fatty acids for both genotypes. The results of this study indicate the potential of using SNPs as DNA markers to select breeding stocks for milk fatty acid composition.<br /> <br /> 10. Experiment station: USDA, ARS, Richard B. Russell Res. Center Agricultural Research Center. Representative: G. J. Hausman. Objectives 1, 2, 3 and 4. The expression of many genes encoding transcription factors, nuclear receptors, enzymes and other regulatory proteins have been studied in human and rodent adipose tissue with cDNA microarrays, but comparable studies in adipose tissue from neonatal and growing pigs have not been reported. In the first study, total RNA was isolated from 90 d fetal stromal-vascular (SV) cell cultures (n = 4; 2 arrays, 2 cultures / array) and adipose tissue from 105 d fetuses (n =2) and neonatal (5 d old) pigs (n=2). In the second study, total RNA was collected at slaughter from outer s.c. adipose tissue (OSQ) and middle s.c. adipose tissue (MSQ) samples from gilts at 90, 150, and 210 d ( n = 5 / age). In both studies, dye-labeled cDNA probes were hybridized to custom microarrays (70 mer oligonucleotides) representing about 600 pig genes involved in growth and reproduction. In the first study, microarray data analysis showed significant expression of 179 genes encoding transcription factors, nuclear receptors, enzymes and other regulatory proteins in fetal SV cell cultures and fetal and neonatal adipose tissue. In the second study, expression intensity ratios revealed little change with age in gene expression overall of 100 transcription factors, nuclear receptors, enzymes and other regulatory proteins in OSQ and MSQ from pigs between 90 - 210 d of age. However, distinct patterns of relative gene expression were evident within adipose tissue from growing pigs (90, 150, and 210 d old pigs). Furthermore, the relative expression of 13 genes distinguished OSQ and MSQ depots in growing pigs. With microarray and RT-PCR assays we showed that the expression of several regulatory genes did not change with age in OSQ and MSQ depots. However, the expression of several genes were influenced by age including an increase in HSF1 and 15-oxoprostaglandin 13-reductase expression in MSQ and a decrease in UCP2 and prohibitin -2 expression in OSQ. These studies demonstrate the expression of several genes in pig adipose tissue that may influence or be associated with leptin expression and adipose tissue metabolism.<br /> Objectives 1, 2 , 3, 4, 6 and 8. In this study, transcriptional profiling was used to identify genetic mechanisms that respond to alpha-MSH, an MC3/4-R agonist. Three MC4R genotypes (2 homozygous and the heterozygous for MC4R) were selected from the UGA swine herd (PIC composite). Thirty six pigs, 6 per genotype per treatment were randomly assigned to one of the following treatments: ICV administration of 150 ul 0.9% saline, or 10 µg NDP-MSH (agonist) in 150 ul of 0.9% saline. Feed intake was measure at 12 and 24 hous after treatment (time 0). All pigs were sacrificed 24 hours post-injection and hypothalamus, liver and middle layer of back fat was collected and mRNA was hybridized to 24,123 probe set Affymetrix Porcine Genome Arrays. MSH suppressed (P< 0.04) feed intake in all animals at 12 and 24 hr after treatment regardless of genotype with no teatment x genotype interaction detected (P > 0.8). A mixed linear model was fit to each tissue and gene using SAS Proc Mixed and interested contrasts were tested. In response to MSH injection, 5070 genes in adipose tissue were declared differentially expressed with <i>q</i>-value<0.07. This criterion was satisfied for 290 adipose tissue genes for the MC4R genotype effect and 1724 adipose genes for the genotype x treatment interaction. Genes representing lipid biosynthesis such as LPL, fatty acid synthase, aconitase-1 and acetyl CoA synthase were down regulated by MSH treatment as were leptin and heat shock protein 70.2 genes. Genes upregulated in adipose tissue tissue by MSH treatment included angiopoietin-like 4, pyruvate dehydrogenase kinase 4 isoform 1, UCP-3 and IGFBP- 3. Angiopoietin-like 4 inhibits LPL activity in rodent adipose tissue and was upregulated more than any other probe in the probe set. Along with blood hormone assays, confirmation of expression of additional candidate genes will be performed to determine key pathways that responded to MSH treatment in pigs. These results demonstrate that the MC4R genotype did not effect the feeding behavior reponse to MSH but did affect adipose tissue gene expression.<br />

Publications

Averette, Gatlin, L., M.T. See, D.K. Larick and J. Odle. 2006. Descriptive flavor analysis of bacon and pork loin from lean-genotype gilts fed conjugated linoleic acid and supplemental fat. J. Anim. Sci. 84:3381-3386.<br /> <br /> Barioni, L. G., J. W. Oltjen, and R. D. Sainz. 2006. Iterative development, evaluation and optimal parameter estimation of a dynamic simulation model: A case study. In: Nutrient Digestion and Utilization in Farm Animals: Modelling Approaches (Kebreab, E., J. Dijkstra, A. Bannink, W. J. J. Gerrits, and J. France, Eds.) pp. 251-256. CAB International, Wallingford, UK.<br /> <br /> Barnes, K.M., and J.L. Miner. 2008. Role of resistin in insulin sensitivity in rodents and humans. Curr. Prot. Peptide Sci. (Accepted.<br /> <br /> Barnes, K.M., and J.L. Miner. 2008. Role of resistin in insulin sensitivity in rodents and humans. Curr. Prot. Peptide Sci. (Accepted.<br /> <br /> Bergen WG. Contributions of research with farm animals to protein metabolism concepts: A historical perspective. 2007. J Nutrition 137: 706-710.<br /> <br /> Bergen WG. Measuring in vivo intracellular protein degradation rates in animal systems. 2007. J Animal Sci.doi:10.2527/jas..2007-0430.<br /> <br /> Blikslager, A.T., A. Moser, J. Gookin, S. Jones and J. Odle. 2007. Restoration of barrier function in injured intestinal mucosa. Physiol. Rev. 87:545-564<br /> <br /> Castro-Bulle, F. C. P., P. V. Paulino, A. C. Sanches, and R. D. Sainz. 2007. Growth, carcass quality, protein and energy metabolism in beef cattle with different growth potentials and residual feed intakes. Journal of Animal Science 85:928-936.<br /> <br /> Corl, B.A., R.J. Harrell, H.K. Moon, O. Phillips, E.M. Weaver, J.M. Campbell, J.A. Arthington and J. Odle. 2007. .Effects of animal plasma proteins on intestinal recovery of neonatal pigs infected with rotavirus. J. Nutr. Biochem. 18:778-784.<br /> <br /> Cummins KA, SG Solaiman, and WG Bergen. The effect of dietary copper supplementation on fatty acid profile and oxidative stability of adipose depots in Boer X Spanish goats. 2007. J Animal Sci. doi: 10.2527<br /> <br /> Deiuliis, J.A., J. Shin, D. Bae, M. J. Azain, R. Barb, and K.Lee. 2008. Developmental, Hormonal, and Nutritional Regulation of Porcine Adipose Triglyceride Lipase (ATGL). Lipids 43:215_25.<br /> <br /> Dicklin, M.E., J.L. Robinson, X. Lin and J. Odle. 2006. Ontogeny and chain-length specificity of gastrointestinal lipases affect medium-chain triacylglycerol utilization by newborn pigs. J. Anim Sci. 84:818-825.<br /> <br /> Dodson, M.V. 2007. Codger and computers: to "unplug" or not to "unplug?" NACTA Journal 51(2):72.<br /> <br /> Dodson, M.V. 2007. In order to recruit animal sciences students into the university, you need to teach them about animal science jobs. NACTA Journal 51(2):72-73. <br /> <br /> Dodson, M.V. 2008. A simplistic view of impact factors: From science to teaching. NACTA Journal<br /> <br /> Dodson, M.V. 2008. Practical ways to measure teaching success. NACTA Journal<br /> <br /> Dodson, M.V. 2008. Alumni associations help teaching efforts on many different levels. NACTA Journal 52(1):66<br /> <br /> Dodson, M.V. 2008. Diversity in academia leads to academic progress. NACTA Journal<br /> <br /> Dodson, M.V. 2008. Funding for enhanced teaching in agriculture. NACTA Journal 52(1):67<br /> <br /> Dodson, M.V. 2008. Relax.....a little, and then move on. NACTA Journal <br /> <br /> Dodson, M.V. and A.M. VanDerZanden. 2008. Contributions of science education journals are enhanced by categorizing (journal) impact factors. NACTA Journal<br /> <br /> Dodson, M.V. and M.E. Fernyhough. 2008. Mature adipocytes: Are there still novel things that we can learn from them? Tissue and Cell [in press]<br /> <br /> Dodson, M.V., A. Kinkel, J.L. Vierck, K. Cain, M. Wick and J. Ottobre. 2008. Undefined cells reside in fish skeletal muscle. Cytotechnology [in press]<br /> <br /> Fernyhough, M.E., E. Okine, G. Hausman, J.L. Vierck and M.V. Dodson. 2007. Invited review: PPAR-gamma and GLUT-4 expression as differentiation markers for preadipocyte conversion to become an adipocyte Domestic Animal Endocrinology 33:367-378<br /> <br /> Fernyhough, M.E., G.J. Hausman and M.V. Dodson. 2008. Progeny from dedifferentiated adipocytes display protracted adipogenesis. Cells, Tissues, Organs [in press]<br /> <br /> Fernyhough, M.E., G.J. Hausman, L.L. Guan, E. Okine, S.S. Moore and M.V. Dodson. 2008. Mature adipocytes may be a source of stem cells for tissue engineering. Biochemical Biophysical Research Communications 368(3):455-457 <br /> <br /> Garcia, F., R. D. Sainz, J. Agabriel and J. W. Oltjen. 2007. Dynamic integration of biological processes into models: contribution to prediction of cattle growth and body composition. In: Ortigues-Marty, I., N. Miraux, W. Brand-Williams, Eds.) Energy and Protein Metabolism and Nutrition. European Federation of Animal Sciencepublication No. 124, pp. 489-490. Wageningen Academic Publishers, Wageningen, The Netherlands.<br /> <br /> Garcia, F., R. D. Sainz, J. Agabriel, L. G. Barioni, and J. W. Oltjen. 2008. Comparative analysis of two dynamic mechanistic models of beef cattle growth. Animal Feed Science and Technology (in press). <br /> <br /> Hargrave, K.M., and J.L. Miner. 2008. Conjugated linoleic acid-induced apoptosis in adipose tissue and 3T3-L1 preadipocytes. Adipocytes. <br /> <br /> Hargrave, K.M., and J.L. Miner. 2008. Conjugated linoleic acid-induced apoptosis in adipose tissue and 3T3-L1 preadipocytes. Adipocytes.<br /> <br /> Hargrave-Barnes, K.M., M.J. Azain, and J.L. Miner. 2008. Conjugated linoleic acid-induced fat loss dependence on D6-desaturase or cyclooxygenase. Obesity .<br /> <br /> Hargrave-Barnes, K.M., M.J. Azain, and J.L. Miner. 2008. Conjugated linoleic acid-induced fat loss dependence on D6-desaturase or cyclooxygenase. Obesity.<br /> <br /> Hausman, G. J., Barb, C. R., and Dean, R. G. Patterns of gene expression in pig adipose tissue: transforming growth factors, interferons, interleukins, and apolipoproteins. J. Anim Sci., 85: 2445-2456, 2007.<br /> <br /> Hausman, G. J., Poulos, S. P., Pringle, T. D., and Azain, M. J. The influence of thiazolidinediones on adipogenesis in vitro and in vivo: Potential modifiers of intramuscular adipose tissue deposition in meat animals. J. Anim Sci., (In press). 2007.<br /> <br /> Hausman, G. J., Barb, C. R., and Dean, R. G. Patterns of gene expression in pig adipose tissue: Insulin-like growth factor system proteins, neuropeptide Y (NPY), NPY receptors, neurotrophic factors and other secreted factors. Domest. Anim Endocrinol.,(In press) 2008. <br /> <br /> Hoppe, Andrea A. and Carey, Gale B. (2007) Polybrominated diphenyl ethers (PBDEs) as endocrine disruptors of adipocyte metabolism. Obesity 15:2942-2950. <br /> <br /> J. Novakofski and S. Brewer. 2006. The Paradox of Toughening During the Aging of Tender Steaks. Journal of Food Science 71(6): S473 479. <br /> <br /> Jones, D. I., F. R. C. Araujo, F. R. B. Ribeiro, M. J. Yokoo, A. C. Sanches, J. W. Oltjen, and R. D. Sainz. 2006. The effects of breed type and growing program on feedlot performance and fat gains in beef steers. Proceedings, Western Section, American Society of Animal Science 57:201-204.<br /> <br /> Kelly A, Mateus Pinilla N, Diffendorfer J, Killefer J, Shelton P, Beissel T, and Novakofski J. 2006. Sequence Variation within the Prion Protein Gene from White tailed Deer (Odocoileus virginianus) of Northern Illinois. Conference for Workers in Animal Diseases.<br /> <br /> Kokta, T.A., A.L. Strat, M.R. Papasani, J. Szasz, M.V. Dodson and R.A. Hill. 2008. Regulation of lipid accumulation in 3T3-L1 cells: Insulin-independent and combined effects of fatty acids and insulin. Animal 2(1):92-99<br /> <br /> Lin, X., B. Corl and J. Odle. 2007. Idiosyncrasies of piglet lipid metabolism and their relationship to postnatal mortality. In: Paradigms in pig science (Edited by J Wiseman et al.) Nottingham University Press. pp 187-206.<br /> <br /> Lkhagvadorj S., Qu L., Cai W., Couture O.P., Wang Y., Barb C.R., Hausman G.J.,Rekaya R., Anderson L.L., Dekkers J.C.M., Nettleton D.S., C.K. Tuggle. 2008. Use of Transcriptional Profiling to Understand Genetic Mechanisms Responding to Fasting in Pigs. Midwest Animal Science Meeting, Des Moines, IA. <br /> <br /> Lkhagvadorj S., Qu L., Cai W., Couture O.P., Wang Y., Barb C.R., Hausman G.J., Rekaya R., Anderson L.L., Dekkers J.C.M., Nettleton D.S., C.d Tuggle. 2008. Sterol Regulatory Transcription Factor-1: Key regulator of Fasting Response in the Adipose Tissue in Pigs?. Experimental Biology Meeting, San Diego, CA. Abstract # 8249 (Accepted)<br /> <br /> Lyvers-Peffer, P.A., X. Lin, S. Jacobi, L.A. Gatlin, J. Woodworth and J. Odle. 2007. Ontogeny of carnitine palmitoyltransferase I activity, carnitine-Km, and mRNA abundance in pigs throughout growth and development. J. Nutr. 137:898-903.<br /> <br /> M. S. Brewer, J. Novakofski and K. Freise. 2006. Instrumental evaluation of pH effects on ability of pork chops to bloom. Meat Science 72( 4):596 602. <br /> <br /> Mateus Pinilla, NE, J Novakofski, J Diffendorfer, J Killefer. Genetic Relatedness and Gene flow of White tailed Deer (Odocoileus virginianus) in the Chronic Wasting Disease prevalent region of Illinois". Illinois Department of Natural Resources and International Association of Fish and Wildlife Agencies (IAFWA). March 2006.<br /> <br /> McPhee, M. J., J. W. Oltjen, J. G. Fadel, D. G. Mayer, and R. D. Sainz. 2007. Parameter Estimation of fat deposition models in beef steers. In Oxley, L. and Kulasiri, D. (eds) MODSIM 2007 International Congress on Modelling and Simulation. Modelling and Simulation Society of Australia and New Zealand, December 2007, pp. 115-120. ISBN : 978-0-9758400-4-7. http:/www.mssanz.au/modsim07/Papers/DegreeofSite_s44_Basenet_.pdf<br /> <br /> McPhee, M. J., J. W. Oltjen, T. R. Famula and R. D. Sainz. 2006. Factors affecting carcass characteristics. California Cattleman 89(9):30-31.<br /> <br /> McPhee, M. J., J. W. Oltjen, T. R. Famula, and R. D. Sainz. 2006. Factors affecting carcass characteristics of feedlot steers: a meta-analysis. Journal of Animal Science 84:3143-3154.<br /> <br /> Meers, S. A., C. R. Dove, and M. J. Azain. 2007. The effect of omega-3 fatty acids on adipose tissue cellularity in grower/finisher pigs. J. Anim. Sci. 85 (Suppl 1): 437.<br /> <br /> Mir, P.S., K. Schwartzhoph-Genswein, E. Okine and M.V. Dodson. 2008. Effect of a short duration feed withdrawal followed by full feeding on marbling fat in beef carcasses. Livestock Science [in press]<br /> <br /> Moutsiolis, Athena A., Rule, Daniel C., Murrieta, Charles M., Bauman, Dale E., Lock, Adam L., Barbano, David M., and Carey, Gale. B. (2008) Human breast milk enrichment in conjugated linoleic acid after consumption of a conjugated linoleic acid-rich food product:: a pilot study. Nutrition Research, in press. <br /> <br /> Novakofski, J. and M. S. Brewer. 2007. Consumer Sensory Evaluations of Aging Effects on Beef Quality," Journal of Food Science (JFS 2007 0542.R1).<br /> <br /> NRC. 1996. Nutrient Requirements of Beef Cattle. 7th rev. ed. National Academy Press, Washington, DC.<br /> <br /> Oltjen, J. W., A. C. Bywater, R. L. Baldwin, and W. N. Garrett. 1986. Development of a dynamic model of beef cattle growth and composition. Journal of Animal Science 62:86-97.<br /> <br /> Oltjen, J. W., R. D. Sainz, A. B. Pleasants, T. K. Soboleva, and V. H. Oddy. 2006. Representation of fat and protein gain at low levels of growth and improved prediction of variable maintenance requirement in a ruminant growth and composition model. In: Nutrient Digestion and Utilization in Farm Animals: Modelling Approaches (Kebreab, E., J. Dijkstra, A. Bannink, W. J. J. Gerrits, and J. France, Eds.) pp. 144-169. CAB International, Wallingford, UK.<br /> <br /> P.J. Oort, C.H. Warden, T.K. Baumann, T.A. Knotts, and S.H. Adams. Characterization of Tusc5, an adipocyte gene co-expressed in peripheral neurons. Mol. Cell. Endocrinol., 276(1-2):24-35, 2007<br /> <br /> P.J. Oort, T.A. Knotts, M. Grino, N. Naour, J-P. Bastard, K. Clément, N. Ninkina, V.L. Buchman, P.A. Permana, X. Luo, G. Pan, T.N. Dunn, and S.H. Adams. ³-Synuclein is an Adipocyte-Neuron Gene Coordinately-Expressed with Leptin and Increased in Human Obesity. in press, J. Nutrition<br /> <br /> Paulino, P .V. R., S. C. Valadares Filho, M. L. Chizzotti, E. Detmann, M. A. Fonseca, M. I. Marcondes and R. D. Sainz. 2007. Net partial efficiencies of metabolizable energy utilization for protein and fat gain in Nellore cattle. In: Ortigues-Marty, I., N. Miraux, W. Brand-Williams, Eds.) Energy and Protein Metabolism and Nutrition. European Federation of Animal Sciencepublication No. 124, pp. 601-602. Wageningen Academic Publishers, Wageningen, The Netherlands.<br /> <br /> Ramsay, T.G., and M. J. Azain. 2007. Comparison of Gene Expression in Lean Contemporary and Crossbred Obese Swine. Adipocyte:2:133-142.<br /> <br /> Reiter SS, CHC Halsey, BM Stronach, JL Bartosh, WF Owsley and WG Bergen. Lipid metabolism related gene-expression profiling in liver, skeletal muscle and adipose tissue in crossbred Duroc and Pietrain pigs. 2007. Comp Biochem Physiol Part D Genomics and Proteomics 2 :200-206.<br /> <br /> Rhoads, J.M., B. Corl, R. Harrell, X. Niu, L. Gatlin, O. Phillips, A.T. Blikslager, A. Moeser, G. Wu and J. Odle. 2007. Intestinal ribosomal p70s6k signaling is increased in piglet rotavirus enteritis. Am. J. Physiol Gastrointest. Liver Physiol 292:G913-G922.<br /> <br /> Rhoads, J.M., X. Niu, J. Odle, and L.M. Graves. 2006. Role of mTOR signaling in intestinal cell migration. Am. J. Physiol Gastrointest. Liver Physiol. 291:G510-G517.<br /> <br /> Sainz, R. D., F. C. P. Castro Bulle, P. R. V. Paulino, and J. F. Medrano. 2007. Residual feed intake, energy and protein metabolism in beef steers. In: Ortigues-Marty, I., N. Miraux, W. Brand-Williams, Eds.) Energy and Protein Metabolism and Nutrition. European Federation of Animal Sciencepublication No. 124, pp. 603-604. Wageningen Academic Publishers, Wageningen, The Netherlands.<br /> <br /> Sainz, R. D., L. G. Barioni, P. V. Paulino, S. C. Valadares Filho, and J. W. Oltjen. 2006. Growth patterns of Nellore vs. British beef cattle breeds assessed using a dynamic, mechanistic model of cattle growth and composition. In: Nutrient Digestion and Utilization in Farm Animals: Modelling Approaches (Kebreab, E., J. Dijkstra, A. Bannink, W. J. J. Gerrits, and J. France, Eds.) pp. 160-170. CAB International, Wallingford, UK.<br /> <br /> Sainz, R.D., and Hasting, E. (2000). Simulation of the development of adipose tissue in beef cattle. In: (McNamara, J. P., J. France, and D. E. Beever, Eds.) Modelling Nutrient Utilization in Farm Animals, pp. 175-182. CAB International, Wallingford, U.K.<br /> <br /> Smith, K.R., S. K. Duckett, M. J. Azain, R. N. Sonon, Jr., and T. D. Pringle. 2007. The effect of anabolic implants on intramuscular lipid deposition in finished beef cattle. J. Anim. Sci. 85: 430-440.<br /> <br /> Taniguchi, M., L. Guan, J. Basarab, M.V. Dodson and S.S. Moore. 2008. Comparative analysis on gene expression profiles in subcutaneous fat tissues of beef cattle. Biochemical Biophysical Research Communications <br /> <br /> Taniguchi, M., L.L. Guan, B. Zhang, M.V. Dodson, E. Okine and S.S. Moore. 2008. Adipogenesis of bovine perimuscular adipocytes. Biochemical Biophysical Research Communications 366:54-59<br /> <br /> Taniguchi, M., L.L. Guan, B. Zhang, M.V. Dodson, E. Okine and S.S. Moore. 2008. Gene expression patterns of bovine perimuscular adipocytes during adipogenesis. Biochemical Biophysical Research Communications 366:346-351<br /> <br /> Tedeschi, L. O., D. G. Fox, R. D. Sainz, L. G. Barioni, S. R. Medeiros, and C. Boin. 2005. Using mathematical models in ruminant nutrition. Scientia Agricola 62:76-91.<br /> <br /> Tsai, T. C., C. R. Dove, M. J. Azain, and M. R. Bedford. 2007. The effect of varied levels of E. Coli phytase on phosphorus balance in weanling pigs. J. Anim. Sci. 85 (Suppl 1.): 156.<br /> <br /> Zumbach, B, I. Misztal, S. Tsurata, J. P. Sanchez, M. J. Azain, W. Herring, J. Holl, and T. Long. 2007. Breeding for robust pigs across the year in heat stress affected areas. J. Anim. Sci. 85 (Suppl. 1): 627.<br /> <br /> <br /> <br />

Impact Statements

1. Improved teaching. MMeting are a unique opportunity for graduate students and postdoctoral fellows to present their research and receive questions and evaluation from senior scientists in an informal setting. Many young scientists clearly find this useful. At the 2008 meeting, six attending committee members had attended NCR97 or NCC097in past years as graduate students, postdoctoral or guests. Five current graduate students and postdoctoral fellows presented research to the group and additional students attended foe the first time without making presentations. The strong attendance from previous students as current members, particularly in light of tight budgets, alternative meetings and increased responsibilities demonstrates the value these scientists place on the educational opportunities presented by the NCCC097 project.
2. Improved swine nutrition. Long-chain PUFAs are precursors for the synthesis of prostanoids. Research demonstrated feeding elevated levels of long-chain PUFAs enhances recovery of ischemic injured porcine ileum and this is mediated via endogenous prostaglandin synthesis.
3. Improved human nutrition. Safety of novel infant formula ingredients were assessed in neonatal pig models. One-day old pigs were fed a cows milk-based formula supplemented with Polydextrose. Digesta pH decreased linearly and quadratically as dietary Polydextrose increased. Polydextrose shows promise as a prebiotic additive in infant formula.
4. Improved human nutrition by improving animal products. Fat-related beef carcass traits are among the major determinants of value in the beef industry. Bovine adipocytes were derived from mature adipocyte dedifferentiation and in analyzed with genomic and proteomic tools.
5. Improved basic knowledge of adipose tissue, potential public health and obesity. The tumor suppressor candidate 5 (Tusc5) highly expressed in adipocytes was described. g-synuclein was found to have similar expression and is significantly increased in adipose of obese humans.
6. Improved swine nutrition and human food products. Adding an omega-3 supplemented to swine diets during growing and/or early finishing phaseincreased muscle content of n-6 fatty acids in pork. There was no effect on growth performance and no obvious fishy odors in the pork. Supplements increased omega-3 content in pork from less than 5 mg per serving of pork to 30-60 mg/serving in 5 to10 weeks.
7. Improved use of by-products for lean animal production. Distillers dried grains with solubles are residue from ethanol production. The calculated iodine value in subcutaneous, belly and leaf fat, but not loin muscle, was increased with feeding of DDGS to swine and addition of hydrogenated fat product to DDGS had minimal effects on fatty acid profiles.
8. Improved growth and efficiency of lean beef. The Davis Growth Model (DGM) is a dynamic, deterministic and mechanistic model of beef cattle growth and body based on fundamental biological concepts of tissue growth as well as net energy. The DGM can be used to accurately forecast body composition over time, but predictions of fat distribution are inadequate without more data on adipose cellularity in animals of different genotypes, ages, and growth trajectories.
9. Improved understanding of body composition determinants. Duroc and Pietrain F1 pigs differ in lipid deposition and fatty acid oxidation. Abundance of mRNA for key lipogenic and beta oxidation enzymes differs between breeds. Commercial beta adrenergic agonist marginally enhanced UCP-3 expression in both pig genotypes.
10. Improved knowledge of the effect of environmental toxins and obesity. Polybrominated diphenyl ethers, a family of flame retardants, are deposited in adipose tissue which becomes more sensitive to epinephrine and less sensitive to insulin.
11. Improved lipid composition of animal foods and human nutrition. Decreasing the percentage of saturated fatty acids in milk can contribute to decreased incidence of CVD in humans. DNA from dairy cows were use to identify a SNP associated with decreased palmitic acid and increased mono- and polyunsaturated fatty which may be used to select breeding stocks with a healthier milk fatty acid composition.
12. Improved knowledge of swine adipose biology. Expression of genes encoding transcription factors, nuclear receptors, enzymes and other regulatory proteins in adipose tissue were examined with cDNA microarrays. 179 genes associated with differences in fetal and neonatal adipose tissue and genes associated with differences in outer subcutaneous and middle adipose tissue were identified.
13. Improved knowledge of food intake regulation. Transcriptional profiling was used to identify genetic mechanisms that respond to an MSH receptor agonist. MC4R genotype did not effect the feeding behavior reponse to MSH but did affect adipose tissue gene expression.
14. Improved basic knowledge of nutrition and obesity. Research examined the effect of oil source on conjugated linoleic acid-induced lipolysis and the effect of DHA on body composition. Dietary CLA was associated with increased basal lipolysis, increased activation of perilipin and de-differentiation of the adipocytes.

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Report Information

Participants

Bergen, Werner (BERGEWG@auburn.edu) - Auburn University; Beitz, Don (dcbeitz@iastate.edu) - Iowa State University; Carey, Gale (gale.carey@unh.edu) - University of New Hampshire; Ajuwon, Kola (kajuwon@purdue.edu)- Purdue University; Hausman, Gary (Gary.Hausman@ars.usda.gov) - USDA ARS, Athens; Adams, Sean (Sean.H.Adams@ARS.USDA.GOV)- USDA ARS, Davis; Barnes, Kimberly (Kim.Barnes@mail.wvu.edu) - West Virginia University; Du, Min (MinDu@uwyo.edu) - University of Wyoming; Schoonmaker, Jon (jschoon@iastate.edu) - Iowa State University; Kinder, James (kinder15@osu.edu)- Ohio State University; Novakofski, Jan (jnova@illinois.edu) - University of Illinois; Brandebourg, Terry (Brandetd@auburn.edu) - Auburn University; Student/guest attendees included Erin Allgood (University of New Hampshire), Siri Ippagunta (West Virginia University), Derris Burnett (Auburn University), Raphael Nafikov (Iowa State University)

Brief Summary of Minutes

Gale Carey volunteered to be secretary for the meeting. There was discussion regarding the location and date of next year's meeting. A poll of those present showed that members preferred meeting at EB (9 in favor, 0 opposed) and preferred meeting before EB starts rather than during EB (7 in favor, 2 opposed). It was agreed that the entire membership should be queried via e-mail, asking individuals to rank their preferences for meeting as either Fri of EB, Sat of EB, Sun of EB, end of EB, or meet at a separate time of year. Kim Barnes and Kola Ajuwon agreed to be co-chairs for next year's meeting.

Cost for this year's meeting is ~$720 for room and lunch. Jan suggested that in the future, the chair negotiate with the hotel for a package deal that includes room and a lunch for attendees, this is much more convenient (final cost per station representative was $65).

Dr. Jim Kinder, Administrative Advisor, reported that our 5-year project renewal was recommended for renewal with minor revisions. Dan Rule has taken the lead on this work, with Gary Hausman's help, input from Harry Mersmann and editing by Jim. Jim reminded us, and his colleagues, that the primary expectation of NCCC097 is meeting to discuss projects and look for synergies, not to conduct collaborative work per se. Revisions that need to be made include: reduce the number of objectives from 10 to less than 5; specify which states are responsible for which objectives; revise objectives so they are different from our previous project; and include milestones with dates; broaden scope to include human obesity. Lastly, all participants must complete Appendix E promptly. Dan Rule will revise the renewal and Don Beitz agreed to assist with this. Revision is due June 1, 2009. Issues of non-attendance were discussed. Members should make every effort to attend annual meeting, and if they do not, their station directors should be notified and they should no longer be members. Members unable to attend each year, but who contribute to the efforts of NCCC097, should be an exception to this guiding principle. Other ideas surfaced including organizing a videoconference for non-attendees, or finding replacements for non-attendees (Texas A&M, Nebraska)? Lastly, Mike Mirando (not present) sent word that Integrated Proposals are a high priority for the USDA, with 4-5 funded last year.

Accomplishments

Auburn University (Bergen, Brandebourg), objectives 1,2,3,4,5,6,7,8,9,10. . The objective of this study was to examine relationships between GE (mRNA abundance) of fatty acid synthase (FAS), PPAR ±, PPAR ³2, carnitine palmitoyl transferase (CPT-1b), leptin, uncoupling protein 2 (UCP-2), ubiquitin conjugating enzyme (E2) and polyubiquitin (PQ) genes in skeletal muscle (SKM) and adipose tissue (AT) with FE and RFI in finishing cattle. Crossbred cattle (bulls, 8; steers, 8) were placed on a feedlot diet and AT and SKM biopsies were taken prior to completion of the finishing trial. Gene expression was determined with real time RT-PCR. Adipose FAS, leptin and PPAR³2 GE were unrelated to RFI. SKM UCP-2 and E-2 but not CPT-1b, PPAR± and PQ GE were correlated with RFI (P<.05). AT leptin GE was higher (P<.05) while SKM UCP-2 GE was lower (P<.05) in bulls than steers. While PQ GE was not correlated with RFI it was highly correlated (P<.01) with E2 GE. Results indicate that AT genes for fat deposition are likely minimally related to RFI and FE, while SKM genes associated with mitochondrial metabolism and protein turnover are more likely correlated with RFI. The transcriptomic signature (GE) for efficiency in beef cattle appears more related to SKM than AT metabolism. A second project examined dopamine effects on adipose tissue. Dopamine (DA) functions both as a neurotransmitter and a hormone through binding to G protein-coupled receptors (DAR) that are classified by their ability to activate (D1R, D5R) or inhibit (D2R, D3R, D4R) adenylate cyclase. However, given the very low serum levels of free DA and lack of information on expression of DAR in human adipose tissue, DA has been ignored as a potential lipolytic agent. Despite this, it is plausible that DA might act peripherally. For instance, over 95% of peripheral DA in humans circulates as biologically inactive DA sulfate (DA-S), which has a long half life (3-4 hrs) compared to 1-2 min for free catecholamines. Furthermore, DA-S is synthesized by the GI tract and its levels increase ~50 fold after meals. Importantly, DA-S can be deconjugated to DA by arylsulfatases, which are expressed in many tissues. A third project's objectives were to: a) examine expression of DAR in visceral (vis) and subcutaneous (sc) adipose tissue, and b) determine whether DA affects adipocyte-specific functions such as lipolysis. Expression of DAR was determined by realtime PCR in pooled vis and sc adipose tissue samples obtained during abdominal surgery. All five DAR isoforms were detected at a relative abundance of: D1>>D2=D4>D3>>D5. This is the first report on expression of functional DAR in adipose tissue and adipocytes. Peripheral DA, originating from the GI tract, could serve as an important link between food intake and lipid metabolism in humans. <br /> <br /> <br /> University of Illinois (Novakofski), objectives 1,2,3,4,5,6,7,8,9,10. Chronic Wasting Disease (CWD), a cervid transmissible spongiform encephalopathy is a neuro-degenerative disease characterized by accumulations of an abnormal form of the prion protein (PrP). Our goal was to identify possible allelic variants related to CWD disease susceptibility. We sequenced Prnp from white-tailed deer in the CWD affected areas of Illinois to determine allelic variation. Tissues were obtained from the Illinois Department of Natural Resources CWD Surveillance program. Animals were selected based on spatial and chronological sampling similarity. We obtained DNA sequences by isolation of DNA followed by amplification of the Prnp gene locus via PCR. Nucleic acid sequences of the prion gene (Prnp) were examined and genotypes compiled for 61 white-tailed deer in northern Illinois, which previously tested positive for Chronic Wasting Disease (CWD), and 121 negative animals selected to control for geographic location and age. Nine nucleotide polymorphisms, seven silent and two coding, were discovered in the sampled population. All observed polymorphisms except one were found in both negative and positive animals, although five polymorphic loci had significantly different distributions of alleles between infected and non-infected individuals. The total number of polymorphisms per animal, silent or coding was negatively correlated to disease status. The potential importance of silent polymorphisms, either individually (60C/T, 153C/T, 555C/T), or cumulatively, in CWD disease sremovtatus has not been previously reported. Microsatellite markers are useful because of their potential to efficiently resolve genetic relationships between individuals in large populations. The objective of this study was to use a panel of microsatellite markers with high-throughput analysis to obtain genetic information in white-tailed deer from Illinois. Using fourteen microsatellites, multi-locus genotypes were compiled for over 700 deer sampled as part of ongoing CWD surveillance in Illinois. Markers were tested for inbreeding patterns and deviations from Hardy-Weinburg equilibrium, indicators of potential bias related to microsatellites. Of the 14 markers, 11 were in Hardy-Weinburg equilibrium and three were associated with inbreeding with in sub-populations. No differences in genetic relatedness were observed between positive and negative animals, suggesting that contact with relatives does not increase the rate of CWD transmission. Within the seven county study area of northern Illinois, female deer exhibited genetic isolation by distance while male deer did not.<br /> <br /> <br /> Iowa State University (Beitz, Schoonmaker, Nafikov), objectives 1,2,3,5,6,9. The overall objective of this research is to develop tools (i.e., DNA markers) that will allow beef producers to enhance the nutritional/health values of beef. To date we have collected approximately 1500 beef samples (out of 2000), analyzed approximately 1050 for triacylglycerol, phospholipid, and total fatty acid composition, sphingomyelin, creatine, creatnine, carnitine, and mineral content. Cholesterol, vitamin E, B6, and B12, and folate content are currently being analyzed. From 2 to 5-fold natural variation exists in all components measured, indicating that genetic selection for altered nutrient content is possible. A second study examined one hundred thirty-seven Angus cross yearling steers (init. BW 390 ± 0.5 kg). Steer were allotted by BW to a 3 ´ 2 factorial arrangement of 6 treatments (4 pens per treatment) to determine the effect of wet distillers grains concentration (0, 20, 40 % diet DM) in high concentrate (12 % hay) and high forage (50 % hay) diets on growth performance and marbling content. Steers were implanted on d 0 with Component TE-S® and were slaughtered in 3 groups when final BW was estimated to be 579 kg. Final BW was similar among treatments and averaged 578 kg. Concentrate-fed steers gained faster (P < 0.01) than did forage-fed steers; amount of distillers grain fed did not affect (P > 0.25) daily gain. Hot carcass weight and dressing percentage was greater (P < 0.01) for concentrate-fed than for forage-fed steers. Dressing percentage tended (P < 0.08) to increase as distillers grain concentration increased. Longissimus dorsi area tended to be greater (P < 0.08) and yield grade was greater (P < 0.01) for concentrate compared with forage-fed cattle. Longissimus dorsi area and yield grade increased (P < 0.03) as distillers grain concentration increased. In concentrate-fed steers, marbling score decreased as distillers grain concentration increased (325, 306, 265), but, in forage-fed steers, marbling score increased from the 0 to 20% inclusion rate, and then decreased from the 20 to 40% inclusion rate (249, 282, 262; diet x distillers grain interaction; P < 0.01). Similarly, in concentrate-fed steers, fat thickness tended to decrease as distillers grain concentration increased, but, in forage-fed steers, fat thickness tended to increase from the 0 to 20% inclusion rate, and then tended to decrease from the 20 to 40% inclusion rate (diet x distillers grain interaction; P < 0.08). In conclusion, when fed to a common live-weight end-point, distillers grain diets alters lean and adipose tissue deposition. <br /> <br /> <br /> University of New Hampshire (Carey, Allgood), objectives 1,4,6,8,9. Our lab conducted 3 projects this past year. The first examined the effect of diet and polybrominated diphenyl ether exposure on adipocyte and whole body metabolism in male Wistar rats. The purpose of this study was to elucidate whether exposure to PBDEs in conjunction with a high-fat/high-sugar diet will exacerbate obesogenic effects in male Wistar rats. Twenty eight rats were fed either a control (C) or high-fat/high-sugar diet (HF), and gavaged with either 18 mg/kg PBDEs (+) or corn oil (-) daily for 4 weeks (n=6-7 per group). Body weight and food intake were measured three times per week. At 3 weeks, 24-hr whole body metabolism was measured. At 4 weeks, blood was sampled for plasma thyroid hormone concentration (T4), epididymal fat pads were removed and weighed, and adipocytes were isolated. Adipocyte size and lipolytic response to adenosine deaminase and varying concentrations of isoproterenol over 90 minutes were measured. Findings revealed that there were no significant differences in lipolysis with ADA or isoproterenol. A significant increase in weight gain was seen in + vs. - rats. HF rats consumed 7.7% more kcals than C rats over the 4 week study (P=0.053). PBDE treatment decreased T4 level by 83% (P<0.05) and tended to decrease insulin levels (P=0.075), compared to control. PBDE administration tended to increase glucose disappearance (P=0.062). PBDE treatment tended to increase energy production (P=0.081). A diet x PBDE treatment interaction (P<0.081) was noted for 4 dependent variables: metabolic efficiency, protein disappearance, epididymal fat pad weight, and insulin level. We conclude that obesogenic effects of PBDEs are modulated by the animal's diet, and that diet should be a key consideration in EDC research. The second project examined the effects of PBDE exposure during pregnancy and lactation on pup growth, food intake, and adipocyte glucose transport. The third project examined the effects of ad libitum feeding on glucose oxidation in isolated adipocytes. <br /> <br /> <br /> North Carolina State University (Odle), objectives 1,2,3,8. The objective of this study was to evaluate the effects of clofibrate on gene expression of hepatic fatty-acid-oxidation and ketogenesis enzymes induced in pigs during neonatal development. Evaluations were conducted in 0, 1, 4 and 7 d-old pigs fed milk replacer and orally gavaged with either vehicle (2 % Tween 80) or clofibrate (75 mg /kg body weight) +/- etomoxir (5 mg/ kg body weight). Transcript abundances were measured using qRT-PCR and were greater for carnitine palmitoyltransfersae I (CPT I; 2.8 fold), carnitine palmitoyltransfersae II (CPT II; 3.2 fold), and mitochondrial 3-methly-3-hydroxyglutaryl-CoA synthase (mHMG-CoA-S; 3.8 fold) in pigs fed clofibrate verses vehicle. Addition of etomoxir had no effects on the transcript abundances induced by clofibrate. Transcript abundance of targeted genes also increased as piglets aged, but the mRNA levels remained relatively constant for CPT I and mHMG-CoA-S in pigs after 4 d and for CPT II after 1 d. There was no interaction between clofibrate treatment and age. The abundance of acyl-CoA oxidase and peroxisome proliferator-activated receptor ± transcripts were not altered by clofibrate, etomoxir or piglet age. In conclusion, clofibrate strongly induces genes of fatty acid oxidation in the young, postnatal pig, but induction is not influenced by developmental age. A second study evaluated the effects of supplementation of ARA on delta-6-desaturase (D6D) and delta-5-desaturase (D5D) mRNA abundance and synthesis of ARA in the intestine and liver. Day old pigs (n=96) were fed a milk based formula for 4, 8, and 16 days. Diets contained either no polyunsaturated fatty acids (0% ARA, negative control), 0.5% ARA, 2.5% ARA, 5% ARA or 5% eicosapentaenoic acid (EPA) of total fatty acids. To measure flux through the desaturase-elongase pathway we incubated liver and intestinal mucosa with 13C-linoleate(C18:2, n-6) and traced its metabolism to ARA via GC/MS. In the intestine accumulation rate (nmol/g tissue/h) of 13C-ARA was not affected by piglet age, but doubled in pigs fed 5%ARA compared with pigs fed 0.5% ARA. Furthermore, accumulation of 13C18:3, n-6 indicated that, among ARA fed pigs, D6D activity was highest in those fed 5% ARA, and this activity was equal to that measured in SRC pigs (P<0.05). In liver, tracer accumulation in ARA was unaffected by age or diet (P>0.05), but accumulation in 13C18:3, n-6 decreased by 38% between d4 and d8. <br /> <br /> <br /> Ohio State University (Lee), objectives 1,2,3,4,5,6,7,8,9,10. Adipose triglyceride lipase (ATGL) is a newly identified lipase. We report for the first time the porcine ATGL sequence and characterize ATGL gene and protein expression in vitro and in vivo. Adult pig tissue expresses ATGL at high levels in the white adipose and muscle tissue relative to other tested tissues. We show that within the white adipose tissue ATGL is expressed at higher levels in the adipocyte than in the stromal-vascular fraction. Additionally, ATGL expression increases dramatically in the subcutaneous adipose during adipose development and maturation, as well as during in vitro adipogenesis. Peroxisome proliferator-activated receptor gamma transcript levels increased concomitant with ATGL gene expression, suggesting a possible role in the regulation of ATGL by adipogenic regulators. In vitro treatment of differentiated primary pig preadipocytes with insulin and forskolin decreased ATGL gene expression in a dose-dependent manner, suggesting ATGL transcript levels are hormone sensitive. In vivo experimentation showed that calorie-restriction in gilts resulted in increased ATGL mRNA and protein levels in subcutaneous and peri-renal fat tissues. Our data demonstrate that ATGL expression reacts to hormonal stimuli and plays a role in catecholamine-induced lipolysis in porcine adipose tissue. In a second study, delta-like protein 1 (DLK1) was examined. Cloning and sequencing of a full length of chicken DLK1 (gDLK1) complementary DNA revealed that gDLK1 contains a total of 1,161 bp, encoding 386 amino acids. The similarity of gDLK1 nucleotide and protein sequences was over 50% compared with other mammalian species. In adipose tissue, the gDLK1 gene was highly expressed in preadipocytes as compared with adipocytes (P < 0.05), whereas expression levels of adipogenic marker genes such as stearoyl-coenzyme A desaturase 1 (SCD-1) and fatty acid binding protein 4 (FABP4) were higher in mature adipocytes than in preadipocytes (P < 0.05 and P < 0.01, respectively). Expression of gDLK1 in adipose tissue tends to decrease with age. The expression of gDLK1 gene in the pectoralis major muscle was significantly higher in 13- and 17-d-old embryos (P < 0.05), decreased in 1- and 5-d-old chicks (P < 0.05), and further decreased in 11- and 33-d-old chickens (P < 0.05). This expression pattern of gDLK1 was very similar to the expression patterns of myogenin and Pax7 genes, suggesting a close association with myogenic activities. In conclusion, the developmental regulation of gDLK1 expression might play an important role in the early stages of adipose and muscle tissue development. <br /> <br /> <br /> Purdue University (Ajuwon), objectives 1,2,3,4,6,8,10. The objective of this study was to investigate the regulation of inflammation by TLR4 and TLR2 ligands in adipocytes. We investigated the response to peptidoglycan from Staphylococcus aureus in differentiated 3T3-L1 adipocytes. Real-time PCR analysis was used to quantify the expression of interleukin 6 (IL6), adiponectin receptors (adipoR1 and adipoR2), toll-like receptor 2 (TLR2) and 4 (TLR2 4). Media level of IL6 was determined with ELISA. RESULTS: Adipocyte stimulation peptidoglycan induces IL6 expression (P < 0.01). Both siRNA mediated suppression of TLR2 and immunoneutralization of TLR2 with a TLR2 specific antibody inhibited response to peptidoglycan.We also examined the regulation of TLR2 and TLR4 mRNA in peptidoglycan treated cells. Both peptidoglycan and lipopolysaccharide (LPS) robustly induce TLR2 mRNA expression, whereas TLR4 mRNA is weakly induced by LPS only. Additionally, peptidoglycan downregulates the mRNA expression of adiponectin receptors, adipoR1 and adipoR2 CONCLUSION: Obesity and type 2 diabetes are associated with increased expression of TLR2, this receptor could play a significant but previously unrecognized role in the establishment of chronic inflammation in adipose tissue in obesity. A second study investigated the metabolic impact of varying the n-3 and n-6 fatty acid concentrations in the diets of male obese Zucker rats fed 5 different diets: Low fat (7 % fat) control diet (C); 4 high fat diets - a saturated fat (25 % beef tallow, diet T), a 20% soy oil (50 % omega-6 linoleic acid, diet S), a 20% fish oil (50 % omega-3 fatty acids from eicosapentaenoic acid, EPA and docosahexaenoic acid, DHA, diet F) and a mixed diet (M) with 10% fish oil and 10 % soybean oil. High fat diets consist of 60 % fat calories relative to control diet (16 % fat calories). A total of eight animals were assigned to each treatment and animals were fed ad libitum for 8 weeks. Animals on the S diet had a higher area under the curve (AUC) for the OGTT relative to other groups, indicating delayed clearance of glucose. The animals on the S diet also had higher total blood cholesterol, free fatty acid and insulin concentrations compared to other groups. Adiposity, measured as percent visceral and epididymal fat pads, was significantly higher in the S group. These animals also had increased basal phosphorylation level of AKT, perhaps as a result of the higher circulating insulin concentration. Animals on the fish oil diet had the lowest adiposity and circulating concentration of free fatty acids and total cholesterol. The 1:1 mixture of soybean oil and fish oil in diet M resulted in metabolic performances that were similar to the control diet. Thus, despite the high lipid intake of the M animals, the 1:1 balance of the n-3 and n-6 fatty acids offsets the negative effects of the high fat intake. <br /> <br /> <br /> Texas A&M University (Smith), objectives 1,2,3,7. We hypothesized L-arginine plus conjugated linoleic acid (CLA) would decrease body fat accumulation by modulating adipose tissue and liver metabolism. Male Sprague Dawley rats were assigned randomly to treatment groups (n = 6 per group): 1) control, fed 2.55% L-alanine plus 1.5% canola oil; 2) arginine, fed 1.25% L-arginine plus 1.5% canola oil; 3) CLA, fed 2.55% L-alanine plus 1.5% CLA; and 4) arginine plus CLA, fed 1.25 % L-arginine plus 1.5% CLA. Daily weight gain, final body weight, and eviscerated body weights were greatest in rats fed arginine plus CLA. The retroperitoneal adipose tissue:body weight ratio was decreased by CLA plus arginine, but epididymal adipose tissue, liver, and soleus and extensor digitorum longus muscle weights were unaffected by arginine or CLA. CLA decreased epididymal adipose tissue concentrations of palmitoleic, oleic and cis-vaccenic acid, and the palmitoleic:stearic acid ratio, indicating an inhibition of stearoyl-CoA desaturase (SCD) activity. CLA increased plasma palmitic and stearic acid, whereas arginine plus CLA increased plasma glycerol. Neither arginine nor CLA affected glucose or palmitate metabolism in liver, but CLA increased glucose carbon incorporation into CO2 and total lipids in epididymal adipose tissue. CLA plus arginine increased palmitate oxidation to CO2 and arginine depressed palmitate incorporation into lipids in adipose tissue. Arginine and CLA decreased most plasma essential amino acids and alanine, glutamate, glutamine, and ornithine. We conclude that arginine stimulated adipose tissue lipolysis and depressed adipose tissue and liver lipid synthesis, whereas CLA, especially in the presence of arginine, may have depressed muscle protein turnover. <br /> <br /> <br /> USDA ARS, Athens (Hausman), objectives 1,2,3,4,5,6,7,8,9,10. Transcriptional profiling was used to identify genes and pathways that responded to intracerebroventricular injection of melanocortin-4 receptor (MC4R) agonist, NDP-MSH in pigs homozygous for MC4R, D298N on allele 1 (n=6) or allele 2 (n=6) or wildtype (, n=6). Feed intake (FI) was measure at 12 and 24 hous after treatment. All pigs were sacrificed 24 hours later and hypothalamus, liver and back fat was collected. NDP-MSH suppressed (p< 0.04) FI at 12 and 24 hr in all animals after treatment except for the 11 genotype MSH treatment which tended to suppress (P = 0.06) feed intake at 12 hr. In response to NDP-MSH, 278 genes in hypothalamus (q£0.07, p£0.001), 249 genes in liver (q£0.07, p£0.001) and 5066 genes in fat (q£0.07, p£0.015), were differentially expressed. Pathway analysis of NDP-MSH induced DE indicated that genes involved in cell communication, nucleotide metabolism, and signal transduction were prominently down regulated in the hypothalamus. In both liver and adipose tissue energy-intensive biosynthetic and catabolic processes were down-regulated in response to NDP-MSH. This included genes encoding biosynthetic pathways such as steroid and lipid biosynthesis, fatty acid synthesis, and amino acid synthesis. Up regulated genes involved direct energy-generating processes such as oxidative phosphorylation, electron transport, ATP synthesis, whereas TCA associated genes were prominently down-regulated in NDP-MSH treated pigs. Our data also indicate a metabolic switch toward energy conservation since genes involved in energy-intensive biosynthetic and catabolic processes were down-regulated in MSH treated pigs.<br /> <br /> <br /> USDA ARS, Davis (Adams), objectives 1,2,4,5,6. Our laboratory focuses on the biology of adipose tissue, with the ultimate goal of applying this knowledge toward strategies to improve public health via nutritional and physical activity interventions. Obesity-related inflammation is also thought to occur in muscle tissue, which may in turn influence muscle metabolic status. Integrins are cell adhesion receptors that mediate cell-cell, cell-ECM, and cell-pathogen interactions and play pivotal roles in leukocyte extravasation, tissue migration, phagocytosis, activation, proliferation, and survival. ²2 integrins are expressed exclusively on leukocytes and consist of four members sharing a common ² subunit, designated CD18, and different ± subunit, designated CD11a through CD11d. CD11d is the most recently identified member of the ²2 integrin subfamily and is expressed primarily on monocytes/macrophages; however, its role in systemic and WAT inflammation associated with excess body fat is poorly understood. We have shown for the first time that CD11d mRNA expression in RWAT is increased dramatically (>300 fold) in obese female Zucker rats compared to non-obese littermates and further confirmed the observation of dramatically increased expression of CD11d in RWAT of obese animals in DIO male mice. To determine whether elevated CD11d expression in RWAT is the result of inflammation associated with increased adiposity or merely general inflammation outside the context of excess body fat, we measured CD11d expression in RWAT of non-obese male mice chronically (5 days) administered a low dose (0.5 mg/kg body weight) the pro-inflammatory stimulus LPS, which has previously been shown to robustly induce WAT inflammation. CD11d expression was increased (~3.6 fold) in RWAT with LPS administration, but far less than observed in obesity despite increased expression of other inflammatory markers generally much greater than observed in obesity. Our results suggest that CD11d may regulate a WAT macrophage subpopulation in obesity consistent with the literature suggesting a more prominent role for this protein in chronic inflammatory conditions rather than acute immune responses.<br /> <br /> <br /> Washington State University (Dodson), objectives 1,3,4,7. During 2008/2009 we have devised methods to isolate and quantitatively evaluate the dedifferentiation of individual mature (lipid-laden) adipocytes from pig-derived (subcutaneous and intramuscular) adipose tissues. We have determined that pig-derived adipocytes must extrude substantial lipid from the cell prior to becoming proliferative-competent. This single observation (of the need to physically rid itself of a majority of cytosolic lipid) is different to what we had previously observed in similar beef cow adipocyte cultures. Significance. We have been criticized that this process may never occur in vivo. Whether it does or doesn't remains to be seen. Instead, this system may be used to identify regulators of the dedifferentiation process, which might (in the future) be exploited for in vivo work. This system (as we have devised it) is novel and our goals are quite simple: to determine the regulation of the potential for adipocytes to either expand their presence in adipose tissue, or to transdifferentiate into other types of cells. In fact, as these cells possess ability to re-proliferate, they may not be traditional adipocytes (as most traditional biologists may have once thought). We are keeping an open mind. Results to support our research. Progeny cells (from beef cows) do NOT express the same gene profile (during a process to re-induce differentiation) as do primary preadipocytes or SV cells (papers published). This work is done in conjunction with researchers in Canada and in China, and is presently funded by a small grant from the University of Alberta.<br /> <br /> <br /> West Virginia University (Barnes), objectives 1,2,10. The primary goal of our laboratory is to understand the mechanism(s) of action of dietary conjugated linoleic acid-induced body fat loss. In the first study, the objectives were to determine if CLA feeding altered adipose tissue lipolysis or the expression of perilipin. Male mice (n=80; 3 wk old) were fed 7% SO or CO diets for 6 wk then 0 or 0.5% CLA for 12 d. A body fat index was calculated: (retroperitoneal+epididymal (EPI) fat pads)*100/body weight. Lipolysis was determined by NEFA and glycerol release from EPI explants. The relative expression of perilipin and phosphorylated perilipin (P-perilipin) were determined by western blotting. The body fat index was reduced by both CLA (P<0.05) and CO (P<0.001) but there was no interaction. NEFA release was increased by CLA in CO-fed mice (2.94 vs 8.63 µmol/g; P<0.05) but not in SO-fed mice (1.76 vs 2.26 µmol/g tissue). Glycerol release was not affected by CO or CLA. Total perilipin was not altered by diet but P-perilipin tended to be increased by CLA in SO-fed mice and decreased by CLA in CO-fed mice (P=0.08). This may indicate that the CLA-stimulated lipolysis in CO-fed mice is on the decline since P-perilipin is associated with increased lipase activity. In the second study, the objectives were to determine if fat accumulates in skeletal muscle of CLA fed mice. Eighty male mice were weaned at 3 weeks of age and randomly allotted to diets: 7% soy oil (SO) or 7% CO and fed for 42 days. For an additional 12 days, half of the mice on each diet were supplemented with 0.5% CLA isomers in place of the basal oil, giving treatments of SO + CLA and CO + CLA, respectively. Feed intake and body weight were measured weekly. At the conclusion of the feeding period mice were killed and retroperitoneal (RP), epididymal (EPI) fat pads, and thigh skeletal muscle were removed and weighed. A body fat index was calculated as ((RP + EPI)/body weight) x 100. Lipid content of skeletal muscle was measured by ether extraction of freeze-dried samples. There were no significant effects of diet on feed intake or body weight. Both CLA (P<0.01) and CO (P<0.01) reduced the body fat index. Therefore, unlike in pigs, CLA fed mice may not accumulate lipid in their skeletal muscle. There may, however, be a difference in skeletal muscle type, as we measured thigh muscle in this study and in most pig studies the longissmus muscles have been analyzed. <br /> <br /> <br /> University of Wyoming (Du), objectives 1,2,3,4,5,6,7,8,9,10. AMP-activated protein kinase (AMPK) has a key role in the regulation of marbling. AMPK activity in skeletal muscle is negatively associated with marbling in beef cattle (Underwood et al., J. Agri. Food Chem. 2007, 55: 9698; Underwood et al., Meat Sci. 2008, 9: 394). Data suggested that nutrition during fetal stage is closely associated with marbling in beef cattle and lamb. We showed that nutrient supplementation during gestation in the dam enhanced adipogenesi in fetal muscle (Tong et al, J. Anim. Sci 2008, 86: 1296; Zhu et al., J. Physiol. 2008, 586: 2651). Maternal obesity induces inflammatory signaling in fetal muscle, which promotes adipogenesis from mesenchymal stem cells. The up-regulation of adipogenesis is associated with the down-regulation of Wnt/b-catenin signaling. <br /> <br />

Publications

Adams, T.H., R. L. Walzem, D. R. Smith, S. Tseng, and S. B. Smith. 2009. Hamburger high in total, saturated and trans-fatty acids decreases HDL cholesterol and increases plasma triacylglycerols by stimulating apparent hepatic de novo lipogenesis and stearoyl-CoA desaturase activity in mildly hypercholesterolemic men. Br. J. Nutr. (Accepted with revision).<br /> <br /> <br /> Adams, S.H., C.L. Hoppel, K.H. Lok, L. Zhao, S.W. Wong, P.E. Minkler, D.H. Hwang, J.W. Newman, and W.T Garvey*. Plasma acylcarnitine profiles suggest incomplete long chain fatty acid ²-oxidation and altered tricarboxylic cycle activity in type 2 diabetic African-American women. J. Nutr. 2009 Apr 15. [Epub ahead of print]<br /> <br /> <br /> Adedokun, Adetayo; Olayiwola Adeola, Michael Sturek, Kolapo Ajuwon. 2009. Obesity Alters Proteoglycan mRNA Abundance in the Subcutaneous Fat Depot of the Ossabaw Miniature Swine. 42nd Midwest Sectional Scientific Sessions. ASAS, 3/16-3/18, 2009, Des Moines, Iowa. <br /> <br /> <br /> Ajuwon KM, Banz W, Winters TA. Stimulation with Peptidoglycan induces interleukin 6 and TLR2 expression and a concomitant downregulation of expression of adiponectin receptors 1 and 2 in 3T3-L1 adipocytes. J Inflamm (Lond). 2009 Apr 6;6:8.<br /> <br /> <br /> Ajuwon, K. M., Winters, T.A. Whisenhunt, B, Banz, W. Alpha-linolenic acid exerts anti-inflammatory effect in 3T3-L1 adipocytes through mechanisms that involve activation of AMPK. Presented at the Joint ASAS-ADSA Meeting, Indianapolis July 7-11, Indianapolis, Indiana.<br /> <br /> <br /> Allen, D. L., and M. Du. (2008). Comparative functional analysis of the cow and mouse myostatin genes reveals novel regulatory elements in their upstream promoter regions. Comparative Biochemistry and Physiology, 150: 432-439.<br /> <br /> <br /> Banz, W.J., H Bryce James, and Kolapo M Ajuwon. Investigating potential anti-diabetic effects of specific soy isoflavones and metabolites in female obese Zucker rats FASEB J. 2008 22:889.10.<br /> <br /> <br /> Barb, C. R., G. J. Hausman, and C. A. Lents. 2008. Effects on neuroendocrine regulation of reproduction in the gilt and sow. 30.<br /> <br /> <br /> Barnes, K.M. and J.L. Miner. 2009. The role of resistin in insulin sensitivity in rodents and humans. Curr. Prot. Pept. Sci. 10:96-107.<br /> <br /> <br /> Barnes, K.M., J.K. Evenson, A.M. Raines, and R.A. Sunde. 2009. Selenium regulation of the selenoproteome in rats. J. Nutr. 139:199-206.<br /> <br /> <br /> Basu, U., M. Taniguchi, Y. Zhao, M.V. Dodson and L.L. Guan. (submitted). Application of 'omics' technologies on improvement of meat quality in livestock species. In Nutritional Biochemistry: Genomics, Metabolomics and Food Supply. Nova Science Publishers, Inc. Hauppauge, NY. [new for 2009]<br /> <br /> <br /> Bergen WG, Wu G. Intestinal Nitrogen Recycling and Utilization in Health and Disease. J Nutr. 2009 Mar 12. [Epub ahead of print].<br /> <br /> <br /> Bergen WG. J Nutr. 2009 Milk-Fat Depression and Lipid Repartitioning in Lactating Dairy Cows. Mar 12. [Epub ahead of print]. <br /> <br /> <br /> Blikslager, A.T., A. Moser, J. Gookin, S. Jones and J. Odle. 2007. Restoration of barrier function in injured intestinal mucosa. Physiol. Rev. 87:545-564<br /> <br /> <br /> Bobe, G., J. A. Minick Bormann, G. L. Lindberg, A. E. Freeman, and D.C. Beitz. 2008. Short Communication: Estimates of genetic variation of milk fatty acids in U.S. Holstein cows. J. Dairy Sci. 91:1209-1213.<br /> <br /> <br /> Broome, A.I., R.J. Harrell, J. Odle, K.E. Sullivan, and J.H. Eisemann. 2008. Determination of the order of limiting amino acids in milk-based liquid diets for pigs from 1.5 to 5.5 kg. American Society of Animal Science Annual Meeting, Indianapolis, IN. <br /> <br /> Brown K.R., R.D. Goodband, M.D. Tokach, S. S. Dritz, J.L. Nelssen, J.E. Minton, J.J. Higgins, X. Lin, J. Odle, J.C. Woodworth, and B.J. Johnson. 2008. Effects of feeding L-carnitine to gilts through day 70 of gestation on litter traits and the expression of insulin-like growth factor system components and L-carnitine concentration in foetal tissues. J. Anim. Physiol. Anim. Nutr. 92:660-667.<br /> <br /> <br /> Burkett, J.L., T.J. Baas, D.C. Beitz, C.R. Schwab, N.L. Berry, and S. Zhang. 2008. Genetic parameter estimates of fatty acid composition and meat quality traits in Duroc pigs selected for intramuscular fat content for six generations. Abst. 116, Amer. Soc. Anim. Sci., Midwest Section, p 37-38.<br /> <br /> <br /> Burnett, Derris D., Lisa A. Kriese Anderson, Dwight F. Wolfe and Werner G. Bergen<br /> Transcriptomic signature of performance efficiency in finishing beef cattle.<br /> EXPO BIOL 2009 (ASN5104).<br /> <br /> <br /> Carey, Gale B (2009) Integrative Metabolism: An interactive learning tool for nutrition, biochemistry and physiology. Journal of College Science Teaching, in press. <br /> <br /> <br /> Chae, S. H., J. T. Keeton, R. K. Miller, D. Johnson, J. Maxim, and S. B. Smith. 2009. The triacylglycerol preparation of conjugated linoleic acid reduces lipid oxidation in irradiated, cooked ground beef patties. Meat Sci. 81:647-652.<br /> <br /> <br /> Chen, J., M. Guridi, M.E. Fernyhough, Z. Jiang, L.L. Guan, E. Okine, G.J. Hausman and M.V. Dodson (resubmitted after revision). Lipid extrusion prior to pig-derived mature adipocyte dedifferentiation. Tissue and Cell [new for 2009]<br /> <br /> <br /> Chen, J., M. Guridi, M.E. Fernyhough, Z. Jiang, L.L. Guan, G.J. Hausman and M.V. Dodson. (submitted). 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Role of leptin in the regulation of growth and carbohydrate metabolism in the ovine fetus during late gestation. Journal of Physiology, 586: 2393-2403.<br /> <br /> <br /> Han, B. , Junfeng Tong, C. Ma, M. J. Zhu, and M. Du. (2008). Insulin-like growth factor-1 (IGF-1) and leucine stimulate mammalian target of rapamycin (mTOR) signaling in pig myogenic satellite cells. Molecular Reproduction and Development, 75: 810-817.<br /> <br /> <br /> Hargrave-Barnes, K.M., M.J. Azain, and J.L. Miner. 2008. Conjugated linoleic acid-induced fat loss dependence on D6-desaturase or cyclooxygenase. Obesity 16:2245-2252.<br /> <br /> <br /> Hausman GJ, Dodson MV, Ajuwon K, Azain M, Barnes KM, Guan LL, Jiang Z, Poulos SP, Sainz RD, Smith S, Spurlock M, Novakofski J, Fernyhough ME, Bergen WG.Board-invited review: the biology and regulation of preadipocytes and adipocytes in meat animals. J Anim Sci. 2009, 87(4):1218-46.<br /> <br /> <br /> Hausman, D. B., H. J. Park, and G. J. Hausman. 2008. 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Jobgen, P. Li, M. J. Lee, S. B. Smith, T. E. Spencer, S. K. Fried, and G. Wu. 2009. Dietary L-arginine supplementation reduces white fat gain and enhances skeletal muscle and brown fat masses in diet-induced obese rats. J. Nutr. 139:230-237.<br /> <br /> <br /> Jobgen, W., W. J. Fu, H. Gao, P. Li, C. J. Meininger, S. B. Smith, T. E. Spencer, and G. Wu. 2009. High fat feeding and dietary L-arginine supplementation differentially regulate gene expression in rat white adipose tissue. Amino Acids 37:187-198. doi:10.1007/s00726-009-0246-7 (e-publication).<br /> <br /> <br /> Kelly, A., N. Mateus-Pinilla, J. Diffendorfer, E. Jewell, M. Ruiz, J. Killefer, P. Shelton, T. Beissel, and J. and Novakofski. 2008. Prion sequence polymorphisms and chronic wasting disease resistance in illinois white-tailed deer (odocoileus virginianus). Prion 2:28-28-36.<br /> <br /> <br /> Kim, Y. H., J. T. Keeton, H. S. Yang, S. B. Smith, J. E. Sawyer, and J. W. Savell. 2009. 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Impact Statements

1. We report for the first time the porcine adipose triglyceride lipase (ATGL) sequence and characterize ATGL gene and protein expression in vitro and in vivo.
2. The developmental regulation of the chicken DLK1 (Delta-like protein 1) expression might play an important role in the early stages of adipose and muscle tissue development.
3. Obesity and type 2 diabetes are associated with increased expression of TLR2, this receptor could play a significant but previously unrecognized role in the establishment of chronic inflammation in adipose tissue in obesity.
4. CD11d may regulate a WAT macrophage subpopulation in obesity consistent with the literature suggesting a more prominent role for this protein in chronic inflammatory conditions rather than acute immune responses.
5. Maternal obesity induces inflammatory signaling in fetal muscle, which promotes adipogenesis from mesenchymal stem cells. The up-regulation of adipogenesis is associated with the down-regulation of Wnt/b-catenin signaling.
6. Within the seven county study area of northern Illinois, female deer exhibited genetic isolation from Chronic Wasting Disease by distance while male deer did not.
7. This is the first report on expression of functional dopamine G-protein coupled receptors (DAR) in adipose tissue and adipocytes. Peripheral dopamine, originating from the GI tract, could serve as an important link between food intake and lipid metabolism in humans.

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