Brief Summary of Minutes of Annual Meeting

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.

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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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.

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). 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] 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. Ajuwon KM, Banz W, Winters TA. 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