RNA-seq reveal role of bovine TORC2 in the regulation of adipogenesis.

Low intramuscular adipose tissue (marbling) continues to be challenge for improving beef quality in Chinese cattle. Highly marbled meat is very desirable; hence, methods to increase IMF content have become a key aspect of improving meat quality. Therefore, research on the mechanism of adipogenesis provides invaluable information for the improvement of meat quality. This study investigated the effect of TORC2 and its underlying mechanism on lipid metabolism in bovine adipocytes. The TORC2 gene was downregulated in bovine adipocytes by siRNA, and RNA sequencing was performed. Downregulation of TORC2 negatively affected bovine adipocyte differentiation. In addition, a total of 577 DEGs were found, containing 146 up-regulated and 376 down-regulated genes. KEGG pathway analysis revealed that the DEGs were linked with neuroactive ligand-receptor interaction pathway, calcium signaling pathway, cAMP pathway, chemokine signaling pathway and Wnt signaling pathway. Gene Ontology (GO) term analysis of the DEGs showed that down-regulation of TORC2 gene significantly suppressed the genes regulating important GO terms of adipogenesis-related processes in bovine adipocytes, especially regulation of biological activity, regulation of primary metabolic process, regulation of multicellular organismal process, cell adhesion, lipid metabolic process, secretion, chemical homeostasis, regulation of transport, cell-cell signaling, cAMP metabolic process, cellular calcium ion homeostasis, fat cell differentiation, and cell maturation. In conclusion, our results suggest that TORC2 at least in part regulates lipid metabolism in bovine adipocytes. The results of this study provide a basis for studying the function and molecular mechanism of the TORC2 gene in regulating adipogenesis.

Maternal Nutrient Restriction and Skeletal Muscle Development: Consequences for Postnatal Health.

Severe undernutrition and famine continue to be a worldwide concern, as cases have been increasing in the past 5 years, particularly in developing countries. The occurrence of nutrient restriction (NR) during pregnancy affects fetal growth, leading to small for gestational age (SGA) or intrauterine growth restricted (IUGR) offspring. During adulthood, SGA and IUGR offspring are at a higher risk for the development of metabolic syndrome. Skeletal muscle is particularly sensitive to prenatal NR. This tissue plays an essential role in oxidation and glucose metabolism because roughly 80% of insulin-mediated glucose uptake occurs in muscle, and it represents around 40% of body weight. Alterations in myofiber number, hypertrophy and myofiber type composition, decreased protein synthesis, lower mitochondrial content and activity of oxidative enzymes, and increased accumulation of intramuscular triglycerides are among the described programming effects of maternal NR on skeletal muscle. Together, these features would add to a phenotype that is prone to insulin resistance, type 2 diabetes, obesity, and metabolic syndrome. Insights from diverse animal models (i.e. ovine, swine, and rodent) have provided valuable information regarding the molecular mechanisms behind those altered developmental pathways. Understanding those molecular signatures supports the development of efficient treatments to counteract the effects of maternal NR on skeletal muscle, and its negative implications for postnatal health.

Supplementary prenatal copper increases plasma triiodothyronine and brown adipose tissue uncoupling protein-1 gene expression but depresses thermogenesis in newborn lambs.

OBJECTIVE: We tested the hypothesis that increasing dietary copper (Cu) to gravid ewes would enhance brown adipose tissue (BAT) thermogenesis in their offspring. METHODS: Twin-bearing ewes were assigned on d 70 of gestation to diets containing 3, 10, or 20 ppm dietary Cu (n = 8 per group). Twin lambs were assigned at birth to a cold (6°C) or warm (28°C) environmental chamber for 48 h. Blood was collected from ewes and from lambs and perirenal BAT was collected after 48 h in the environmental chambers. RESULTS: Prenatal Cu exposure increased ewe plasma triiodothyronine (T3) and thyroxine concentration (T4) (p < 0.01) but prenatal Cu exposure had no effect on lamb plasma concentrations of T3, T4, glucose, or nonesterified fatty acid concentration (p ≥ 0.08). The high level of prenatal Cu exposure depressed 48-h rectal temperature (p = 0.03). Cold exposure decreased BAT norepinephrine (NE) and increased BAT dopamine (p ≤ 0.01), but prenatal Cu exposure had no effect on BAT cytochrome C oxidase activity or BAT NE or dopamine (p ≥ 0.07). However, BAT of lambs from high-Cu ewes maintained higher uncoupling protein-1 (UCP1) gene expression than BAT of lambs from low- and medium-Cu ewes following warm or cold exposure in environmental chambers (p = 0.02). Cold exposure caused near depletion of BAT lipid by 48 h (p < 0.001), increased BAT cytochrome c oxidase activity (p < 0.01), and depressed plasma fatty acid concentrations (p < 0.001). CONCLUSION: Although prenatal Cu exposure increased BAT UCP1 expression during warm and cold exposure, prenatal cold Cu exposure depressed 48-h rectal temperature. Cold exposure decreased BAT lipid content by over 80% and decreased lamb plasma fatty acid concentration by over 40%, indicating that fuel reserves for thermogenesis were nearly depleted by 48 h of cold exposure.

Genetic Architecture and Selection of Chinese Cattle Revealed by Whole Genome Resequencing.

The bovine genetic resources in China are diverse, but their value and potential are yet to be discovered. To determine the genetic diversity and population structure of Chinese cattle, we analyzed the whole genomes of 46 cattle from six phenotypically and geographically representative Chinese cattle breeds, together with 18 Red Angus cattle genomes, 11 Japanese black cattle genomes and taurine and indicine genomes available from previous studies. Our results showed that Chinese cattle originated from hybridization between Bos taurus and Bos indicus. Moreover, we found that the level of genetic variation in Chinese cattle depends upon the degree of indicine content. We also discovered many potential selective sweep regions associated with domestication related to breed-specific characteristics, with selective sweep regions including genes associated with coat color (ERCC2, MC1R, ZBTB17, and MAP2K1), dairy traits (NCAPG, MAPK7, FST, ITFG1, SETMAR, PAG1, CSN3, and RPL37A), and meat production/quality traits (such as BBS2, R3HDM1, IGFBP2, IGFBP5, MYH9, MYH4, and MC5R). These findings substantially expand the catalogue of genetic variants in cattle and reveal new insights into the evolutionary history and domestication traits of Chinese cattle.

Current situation and future prospects for global beef production: overview of special issue.

The demand for beef as a protein source is increasing worldwide, although in most countries beef accounts for considerably less than half of total meat consumption. Beef also provides a highly desirable eating experience in developed countries and, increasingly, in developing countries. The sustainability of beef production has different meanings in the various geographical and socio-economic regions of the world. Natural resources including land mass and uses, rainfall and access to livestock feed, and the robustness of the economy are major determinants of the perception of beef sustainability. In this overview of the 2016 International Symposium on "Future Beef in Asia" and this subsequent Special Edition of the Asian-Australasian Journal of Animal Sciences on "Current Situation and Future Prospects for Global Beef Production", the contributions have been grouped into the following categories: Countries in Southeast Asia; Europe; and Countries producing highly marbled beef for export and/or domestic consumption. They also include reference to Special Topics including marbled beef production, and use of "omics" technologies to enhance beef quality assurance. Among these broad categories, notable differences exist across countries in the production and marketing of beef. These reflect differences in factors including natural resource availability and climate, population size, traditional culture and degree of economic development including industrial and technological developments. We trust that the International Symposium and this Special Edition on Current Situation and Future Prospects for Global Beef Production, the contents of which that are briefly summarized in this paper, will serve as a valuable resource for the livestock industries, researchers and students with an interest in enhancing the prospects for sustainable, efficient beef production that satisfies the growing size and complexity of consumer demands and markets for beef.

The Expression of Adipogenic Genes in Adipose Tissues of Feedlot Steers Fed Supplementary Palm Oil or Soybean Oil.

We hypothesized that supplementing finishing diets with palm oil would promote adipogenic gene expression and stearoyl-CoA desaturase (SCD) gene expression in subcutaneous (s.c.) and intramuscular (i.m.) adipose tissues of feedlot steers. Eighteen Angus and Angus crossbred steers were assigned to three groups of 6 steers and fed a basal diet (control), with 3% palm oil, or with 3% soybean oil, for 70 d, top-dressed daily. Tailhead s.c. adipose tissue was obtained by biopsy at 14 d before the initiation of dietary treatments and at 35 d of dietary treatments. At slaughter, after 70 d of dietary treatment, tailhead s.c. adipose tissue and i.m. adipose tissue were obtained from the longissimus thoracis muscle. Palm oil increased plasma palmitic acid and soybean oil increased plasma linoleic acid and α-linolenic acid relative to the initial sampling time. Expression of AMP-activated protein kinase alpha (AMPKα) and peroxisome proliferator-activated receptor gamma (PPARγ) increased between the initial and intermediate biopsies and declined thereafter (p<0.03). SCD gene expression did not change between the initial and intermediate biopsies but declined by over 75% by the final period (p = 0.04), and G-coupled protein receptor 43 (GPR43) gene expression was unaffected by diet or time on trial. Soybean oil decreased (p = 0.01) PPARγ gene expression at the intermediate sample time. At the terminal sample time, PPARγ and SCD gene expression was less in i.m. adipose tissue than in s.c. adipose tissue (p<0.05). AMPKα gene expression was less in s.c. adipose tissue of palm oil-fed steers than in control steers (p = 0.04) and CCAAT enhancer binding protein-beta (CEBPß) gene expression was less in s.c. and i.m. adipose tissues of palm oil-fed steers than in soybean oil-fed steers (p<0.03). Soybean oil decreased SCD gene expression in s.c. adipose tissue (p = 0.05); SCD gene expression in palm oil-fed steers was intermediate between control and soybean oil-fed steers. Contrary to our original hypothesis, palm oil did not promote adipogenic gene expression in s.c. and i.m. adipose tissue.

The dimeric structure of factor XI and zymogen activation.

Factor XI (fXI) is a homodimeric zymogen that is converted to a protease with 1 (1/2-fXIa) or 2 (fXIa) active subunits by factor XIIa (fXIIa) or thrombin. It has been proposed that the dimeric structure is required for normal fXI activation. Consistent with this premise, fXI monomers do not reconstitute fXI-deficient mice in a fXIIa-dependent thrombosis model. FXI activation by fXIIa or thrombin is a slow reaction that can be accelerated by polyanions. Phosphate polymers released from platelets (poly-P) can enhance fXI activation by thrombin and promote fXI autoactivation. Poly-P increased initial rates of fXI activation 30- and 3000-fold for fXIIa and thrombin, respectively. FXI monomers were activated more slowly than dimers by fXIIa in the presence of poly-P. However, this defect was not observed when thrombin was the activating protease, nor during fXI autoactivation. The data suggest that fXIIa and thrombin activate fXI by different mechanisms. FXIIa may activate fXI through a trans-activation mechanism in which the protease binds to 1 subunit of the dimer, while activating the other subunit. For activation by thrombin, or during autoactivation, the data support a cis-activation mechanism in which the activating protease binds to and activates the same fXI subunit.

A pilot study on the effect of nasal continuous positive airway pressure on arterial partial pressure of carbon dioxide during spinal anesthesia with intravenous sedation for total knee arthroplasty.

BACKGROUND: Deep sedation of surgical patients may be associated with hypoventilation, airway collapse, and hypercarbia, although the extent of hypercarbia is rarely quantified. In this prospective, randomized, controlled clinical pilot study, we assessed the efficacy of nasal continuous positive airway pressure (nCPAP) for reducing arterial partial pressure of carbon dioxide (PaCO2) among deeply sedated, spontaneously ventilated patients undergoing total knee arthroplasty (TKA) under subarachnoid block (SAB), versus standard airway management in a control group. METHODS: Forty ASA status I-III patients underwent deep sedation with propofol to level 2 on the Modified Observers Assessment of Alertness/Sedation Scale during TKA performed under SAB. Nasal or oral airways were placed at the discretion of the anesthesia team, but they were not used in conjunction with nCPAP. Baseline arterial blood gas analysis (ABG-1) was performed after Modified Observers Assessment of Alertness/Sedation Scale level 2 was reached. Patients were then randomized to receive nCPAP (nCPAP group, N = 20) or standard oxygen mask management (control group, N = 20). A second ABG (ABG-2) was performed 30 minutes later to assess the effect of nCPAP on PaCO2. The primary efficacy end point was change in PaCO2 from baseline to the 30-minute time point. RESULTS: Baseline (ABG-1) PaCO2 values were similar between nCPAP and control groups with median values of 54.5 and 56.1 mm Hg, respectively. There was a significant decline in PaCO2 in the nCPAP group (median of -4.6 mm Hg [10th-90th quantile, -14.55 to 3.85]) as compared with the control group (median of 0.95 mm Hg [-4.75 to 9.85]; P = 0.015; 95% confidence interval [CI] for location shift = -9.5 to -1.3). Within the control group, PaCO2 was similar from ABG-1 to ABG-2 (median [10th-90th quantile] = 56.1 mm Hg [47.2-67.0] vs 56.6 mm Hg [46-68.8]; P = 0.52; 95% CI for the median = -3.4 to 3.4). Forty percent of all patients received an airway before ABG-1. The baseline PaCO2 value of patients receiving an airway was not different from that of patients without an airway (median [10th-90th quantile] = 56.0 mm Hg [46.0-68.4] vs 54.1 mm Hg [45.6-65.6], respectively; P = 0.33; 95% CI for location shift = -2.30 to 7.20). CONCLUSIONS: Deep sedation of TKA patients during SAB resulted in moderate hypercarbia (mean and median PaCO2 = 55). There was a trend showing that nCPAP treatment reduced PaCO2 versus treatment for control group patients receiving standard airway management; however, estimated treatment difference varied widely, from 1.4 to 12.6 mm Hg. Among control group patients, the initial PaCO2 during deep sedation was similar to the PaCO2 when measured after a 30-minute period of continued deep sedation. Finally, baseline PaCO2 among deeply sedated patients who received an airway was not different from that of patients who did not receive an airway.

Abomasal infusion of arginine stimulates SCD and C/EBPß gene expression, and decreases CPT1ß gene expression in bovine adipose tissue independent of conjugated linoleic acid.

Based on previous research with bovine peadipocytes, we hypothesized that infusion of arginine into the abomasum of Angus steers stimulates stearoyl-CoA desaturase (SCD) gene expression in bovine subcutaneous (s.c.) adipose tissue, and that this would be attenuated by conjugated linoleic acid (CLA). Growing Angus steers were infused abomasally with L-arginine 50 g/day; n = 13; provided as L-arginine HCl) or L-alanine (isonitrogenous control, 100 g/day; n = 11) for 14 days. For the subsequent 14 days, half of the steers in each amino acid group were infused with CLA (100 g/day). Body weight gain and average daily gain were unaffected (P > 0.15) by infusion of arginine or CLA into the abomasum. The plasma concentrations of cis-9, trans-11 and trans-10, cis-12 CLA were increased CLA infusion (P = 0.001) and infusion of arginine increased plasma arginine (P = 0.01). Compared with day 0, fatty acid synthase, glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase enzyme activities in s.c. adipose tissue increased by day 14 in steers infused with either alanine or arginine (all P < 0.01). NADP-MDH activity was higher (P = 0.01) in steers infused with arginine than in steers infused with arginine plus CLA by day 28, but lipid synthesis in vitro from glucose and acetate was unaffected by infusion of either arginine or CLA (P > 0.40). By day 28, C/EBPß and SCD gene expression was higher, and CPT1ß gene expression was lower, in s.c. adipose tissue of steers infused with arginine than in steers infused with alanine (±CLA) (P = 0.05). CLA decreased adipose tissue oleic acid (18:1n-9) in alanine- or arginine-infused steers (P = 0.05), although CLA had no effect on SCD gene expression. The data indicate that supplemental arginine promotes adipogenic gene expression and may promote lipid accumulation in bovine adipose tissue. L-Arginine may beneficially improve beef quality for human consumption.

Inhibition of Escherichia coli O157:H7 and Salmonella enterica on spinach and identification of antimicrobial substances produced by a commercial Lactic Acid Bacteria food safety intervention.

The microbiological safety of fresh produce is of concern for the U.S. food supply. Members of the Lactic Acid Bacteria (LAB) have been reported to antagonize pathogens by competing for nutrients and by secretion of substances with antimicrobial activity, including organic acids, peroxides, and antimicrobial polypeptides. The objectives of this research were to: (i) determine the capacity of a commercial LAB food antimicrobial to inhibit Escherichia coli O157:H7 and Salmonella enterica on spinach leaf surfaces, and (ii) identify antimicrobial substances produced in vitro by the LAB comprising the food antimicrobial. Pathogens were inoculated on freshly harvested spinach, followed by application of the LAB antimicrobial. Treated spinach was aerobically incubated up to 12 days at 7 °C and surviving pathogens enumerated via selective/differential plating. l-Lactic acid and a bacteriocin-like inhibitory substance (BLIS) were detected and quantified from cell-free fermentates obtained from LAB-inoculated liquid microbiological medium. Application of 8.0 log10 CFU/g LAB produced significant (p < 0.05) reductions in E. coli O157:H7 and Salmonella populations on spinach of 1.6 and 1.9 log10 CFU/g, respectively. It was concluded the LAB antimicrobial inhibited foodborne pathogens on spinach during refrigerated storage, likely the result of the production of metabolites with antimicrobial activity.

Historical Overview of the Effect of ß-Adrenergic Agonists on Beef Cattle Production.

Postnatal muscle hypertrophy of beef cattle is the result of enhanced myofibrillar protein synthesis and reduced protein turnover. Skeletal muscle hypertrophy has been studied in cattle fed ß-adrenergic agonists (ß-AA), which are receptor-mediated enhancers of protein synthesis and inhibitors of protein degradation. Feeding ß-AA to beef cattle increases longissimus muscle cross-sectional area 6% to 40% compared to non-treated cattle. The ß-AA have been reported to improve live animal performance, including average daily gain, feed efficiency, hot carcass weight, and dressing percentage. Treatment with ß-AA increased mRNA concentration of the ß 2 or ß 1-adrenergic receptor and myosin heavy chain IIX in bovine skeletal muscle tissue. This review will examine the effects of skeletal muscle and adipose development with ß-AA, and will interpret how the use of ß-AA affects performance, body composition, and growth in beef cattle.

A sequential mechanism for exosite-mediated factor IX activation by factor XIa.

During blood coagulation, the protease factor XIa (fXIa) activates factor IX (fIX). We describe a new mechanism for this process. FIX is cleaved initially after Arg(145) to form fIXα, and then after Arg(180) to form the protease fIXaß. FIXα is released from fXIa, and must rebind for cleavage after Arg(180) to occur. Catalytic efficiency of cleavage after Arg(180) is 7-fold greater than for cleavage after Arg(145), limiting fIXα accumulation. FXIa contains four apple domains (A1-A4) and a catalytic domain. Exosite(s) on fXIa are required for fIX binding, however, there is lack of consensus on their location(s), with sites on the A2, A3, and catalytic domains described. Replacing the A3 domain with the prekallikrein A3 domain increases K(m) for fIX cleavage after Arg(145) and Arg(180) 25- and ≥ 90-fold, respectively, and markedly decreases k(cat) for cleavage after Arg(180). Similar results were obtained with the isolated fXIa catalytic domain, or fXIa in the absence of Ca(2+). Forms of fXIa lacking the A3 domain exhibit 15-fold lower catalytic efficiency for cleavage after Arg(180) than for cleavage after Arg(145), resulting in fIXα accumulation. Replacing the A2 domain does not affect fIX activation. The results demonstrate that fXIa activates fIX by an exosite- and Ca(2+)-mediated release-rebind mechanism in which efficiency of the second cleavage is enhanced by conformational changes resulting from the first cleavage. Initial binding of fIX and fIXα requires an exosite on the fXIa A3 domain, but not the A2 or catalytic domain.

Sox4 cooperates with PU.1 haploinsufficiency in murine myeloid leukemia.

Cooperation of multiple mutations is thought to be required for cancer development. In previous studies, murine myeloid leukemias induced by transducing wild-type bone marrow progenitors with a SRY sex determining region Y-box 4 (Sox4)-expressing retrovirus frequently carried proviral insertions at Sfpi1, decreasing its mRNA levels, suggesting that reduced Sfpi1 expression cooperates with Sox4 in myeloid leukemia induction. In support of this hypothesis, we show here that mice receiving Sox4 virus-infected Sfpi1(ko/+) bone marrow progenitors developed myeloid leukemia with increased penetrance and shortened latency. Interestingly, Sox4 expression further decreased Sfpi1 transcription. Ectopic SOX4 expression reduced endogenous PU.1 mRNA levels in HL60 promyelocytes, and decreased Sfpi1 mRNA levels were also observed in the spleens of leukemic and preleukemic mice receiving Sox4 virus-infected wild-type bone marrow cells. In addition, Sox4 protein bound to a critical upstream regulatory element of Sfpi1 in ChIP assays. Such cooperation probably occurs in de novo human acute myeloid leukemias, as an analysis of 285 acute myeloid leukemia patient samples found a significant negative correlation between SOX4 and PU.1 expression. Our results establish a novel cooperation between Sox4 and reduced Sfpi1 expression in myeloid leukemia development and suggest that SOX4 could be an important new therapeutic target in human acute myeloid leukemia.

Development of a postexposure biologic treatment algorithm on a medical center campus.

OBJECTIVE: To describe the development of an easily accessible online biologic exposure algorithm to guide postexposure medical evaluation and treatment of medical research personnel and health care workers in a Midwest medical center campus. METHODS: We describe the steps involved in the creation of a biologic exposure algorithm from design through implementation. RESULTS: One point of contact allows phone evaluation and immediate triage, providing effective and timely medical care for exposed employees as well as important guidance for clinicians. The algorithm and exposure response system achieved the goal of integrating clinical and research laboratory exposure response. CONCLUSIONS: Development of an integrated clinical and research exposure protocol may be an efficient way to maximize biosafety for workers.

GPR120 Gene expression and activity in subcutaneous and intramuscular adipose tissues of Angus crossbred steers.

We hypothesized that media long-chain fatty acids (LCFA) would more greatly depress cyclic adenosine monophosphate (cAMP), glycerol, and free fatty acid (FFA) concentrations in subcutaneous (s.c.) adipose tissue than in intramuscular (i.m.) adipose tissue via G protein-coupled receptor 120 (GPR120). The GPR120 receptor binds to LCFA, which reduces cAMP production, thereby causing a depression in lipolysis. Fresh ex vivo explants of s.c. and i.m. adipose tissue from the fifth to eighth longissimus thoracic rib muscle section of 8, 22-mo-old Angus crossbred steers were transferred immediately to 6-well culture plates containing 3 mL of Krebs-Henseleit buffer/Hepes/5 mM glucose. Samples were preincubated with 0.5 mM theophylline plus 10 µM forskolin for 30 min, after which increasing concentrations of acetate or propionate (volatile fatty acids, VFA) (0, 1, 5, and 10 mM) in the absence or presence of 100 µM oleate (18:1n-9) or 100 µM palmitate (16:0) (LCFA) were added to the incubation media and incubated an additional 30 min. Main effects of adipose tissue depot (i.m. vs. s.c) and VFA (acetate vs. propionate) for adipose tissue concentrations of forskolin-stimulated cAMP were P = 0.747 and P = 0.106, respectively. The addition of LCFA to the media depressed adipose tissue concentrations of cAMP (P = 0.006) (LCFA main effects). The Tissue × VFA × LCFA interaction was not significant for any dependent variable (P ≥ 0.872). Therefore, concentrations of cAMP, glycerol, and FFA were analyzed separately for i.m. and s.c. adipose tissue by split-plot analysis. Concentrations of cAMP, glycerol, or FFA in i.m. and s.c. adipose tissue were not affected by increasing concentrations of VFA (P ≥ 0.497). Media LCFA had no effect on i.m. adipose tissue cAMP (P = 0.570) or glycerol (P = 0.470) but depressed i.m. adipose tissue FFA (P < 0.001). In s.c. adipose tissue, LCFA decreased concentrations of cAMP (P = 0.042) and glycerol (P = 0.038), but increased FFA concentration (P = 0.026). Expression of GPR120 (P = 0.804) and stearoyl-CoA desaturase (P = 0.538) was not different between s.c. adipose tissue and i.m. adipose tissue. The binding of VFA to the GPR43 receptor depresses cAMP production, thereby attenuating lipolysis, but GPR43 mRNA was undetectable in those adipose tissue samples. These results provide evidence for functional GPR120 receptors in s.c. adipose tissue but question the role of GPR43 in the accumulation of adipose tissue lipids in growing steers.

We measured the mRNA abundance and activity of the fatty acid receptor, G protein-coupled receptor 120 (GPR120) in bovine subcutaneous and intramuscular (marbling) adipose tissue. The GPR120 receptor binds to long-chain fatty acids, which reduces cyclic adenosine monophosphate (cAMP) production, thereby decreasing lipolysis. The mRNA amount of GPR120 was similar between subcutaneous and intramuscular adipose tissues. In subcutaneous and intramuscular adipose tissue incubated in vitro, the fatty acids oleic acid and palmitic acid (the most abundant fatty acids in bovine adipose tissue) strongly depressed the production of cAMP and glycerol in subcutaneous adipose tissue and decreased the concentration of free fatty acids in intramuscular adipose tissue (all measured with commercial kits). This indicates that elevations in adipose tissue or plasma fatty acids may promote fat accumulation by decreasing the breakdown of stored lipids via GPR120. The volatile fatty acids acetate and propionate, which bind to G protein-coupled receptor 43 (GPR43) had no effect on cAMP, glycerol, or free fatty acids. This questions the role of GPR43 in the accumulation of adipose tissue lipids in growing steers.

Vitamin D3 decreases myoblast fusion during the growth and increases myogenic gene expression during the differentiation phase in muscle satellite cells from Korean native beef cattle.

The process of myogenesis, which involves the growth and differentiation of muscle cells, is a crucial determinant of meat yield and quality in beef cattle. Essential nutrients, such as vitamins D and A, play vital roles in the development and maintenance of various tissues, including muscle. However, limited knowledge exists regarding the specific effects of vitamins A and D in bovine muscle. Therefore, the aim of this study was to investigate the impact of vitamins A and D treatment on myogenic fusion and differentiation in bovine satellite cells (BSC). BSC were isolated from Korean native beef cattle, specifically from four female cows approximately 30 mo old. These individual cows were used as biological replicates (n = 3 or 4), and we examined the effects of varying concentrations of vitamins A (All-trans retinoic acid; 100 nM) and D (1,25-dihydroxy-vitamin D3; 1 nM, 10 nM, and 100 nM), both individually and in combination, on myoblast fusion and myogenic differentiation during the growth phase (48 h) or differentiation phase (6 d). The results were statistically analyzed using GLM procedure of SAS with Tukey's test and t-tests or one-way ANOVA where appropriate. The findings revealed that vitamin A enhanced the myoblast fusion index, while vitamin D treatment decreased the myoblast fusion index during the growth phase. Furthermore, vitamin A treatment during the differentiation phase promoted terminal differentiation by regulating the expression of myogenic regulatory factors (Myf5, MyoD, MyoG, and Myf6) and inducing myotube hypertrophy compared to the control satellite cells (P < 0.01). In contrast, vitamin D treatment during the differentiation phase enhanced myogenic differentiation by increasing the mRNA expression of MyoG and Myf6 (P < 0.01). Moreover, the combined treatment of vitamins A and D during the growth phase increased myoblast fusion and further promoted myogenic differentiation and hypertrophy of myotubes during the differentiation phase (P < 0.01). These results suggest that vitamin A and D supplementation may have differential effects on muscle development in Korean native beef cattle during the feeding process.

The study investigated the effects of vitamins A and D on the growth and differentiation phases of bovine satellite cells and found that both vitamins have a positive impact on muscle development. Vitamin A promoted myoblast fusion during the growth phase, leading to increased myotube formation, while vitamin D suppressed myoblast fusion during this phase. However, during the differentiation phase, both vitamins enhanced terminal differentiation and hypertrophy. Vitamin A promoted the activation of satellite cells, while vitamin D promoted the expression of genes that enhance myogenesis. The combination treatment of vitamins A and D during the growth phase complemented each other to increase myogenic cell fusion, and during differentiation, promoted terminal differentiation and hypertrophy. These findings suggest that supplementing cattle feed with both vitamins A and D has the potential to enhance muscle development, which would be advantageous for the meat industry.

Consuming High-Fat and Low-Fat Ground Beef Depresses High-Density and Low-Density Lipoprotein Cholesterol Concentrations, and Reduces Small, Dense Low-Density Lipoprotein Particle Abundance.

We hypothesized that consumption of high-fat (HF) ground beef (24% fat) would not affect plasma concentrations of high-density lipoprotein cholesterol (HDL-C) or low-density lipoprotein (LDL-C), whereas low-fat (LF) ground beef (5% fat) would decrease HDL-C and LDL-C concentrations. In a randomized 2-period crossover, controlled feeding trial, 25 men (mean age and body mass index, 40 years and 31.2) consumed 115-g HF or LF patties, 5/week for 5 weeks with a 4-week washout. The HF treatment increased % energy from fat (p = 0.006) and saturated fat (p = 0.004) and tended (p = 0.060) to depress % energy from carbohydrates. The HF and LF treatments decreased the plasma concentrations of HDL-C (p = 0.001) and LDL-C (p = 0.011). Both ground beef treatments decreased the abundance of HDL3a and increased the abundance of HDL3 (p ≤ 0.003); the LF treatment also decreased the abundance of HDL2b and HDL2a (p ≤ 0.012). The HF and LF treatments decreased the abundance of LDL3 and LDL4 (p ≤ 0.024) and the HF treatment also decreased LDL5 (p = 0.041). Contrary to our hypothesis, the HF treatment decreased plasma HDL-C and LDL-C concentrations despite increased saturated fat intake, and both treatments decreased the abundance of smaller, denser LDL subfractions.

Assessment of Vascular Function in Response to High-Fat and Low-Fat Ground Beef Consumption in Men.

Red meat is stigmatized as an unhealthy protein choice; however, its impacts on vascular function have not been evaluated. We aimed to measure the vascular impact of adding either low-fat (~5% fat) ground beef (LFB) or high-fat (~25% fat) ground beef (HFB) to a habitual diet in free-living men. Twenty-three males (39.9 ± 10.8 years, 177.5 ± 6.7 cm, 97.3 ± 25.0 kg) participated in this double-blind crossover study. Assessment of vascular function and aerobic capacity were measured at entry and in the last week of each intervention and washout period. Participants then completed two 5-week dietary interventions (LFB or HFB; 5 patties/week) in a randomized order with a 4-week washout. Data were analyzed via 2 × 2 repeated-measures ANOVA (p < 0.05). The HFB intervention improved FMD relative to all other time points, while lowering systolic (SBP) and diastolic blood pressure (DBP) relative to entry. Neither the HFB nor the LFB altered pulse wave velocity. The addition of either low- or high-fat ground beef did not negatively alter vascular function. In fact, consuming HFB improved FMD and BP values, which may be mediated by lowering LDL-C concentrations.

Targeted metabolomics characterizes metabolite occurrence and variability in stable freshwater mussel populations.

Freshwater mussels (order Unionida) play a key role in freshwater systems as ecosystem engineers and indicators of aquatic ecosystem health. The fauna is globally imperilled due to a diversity of suspected factors; however, causes for many population declines and mortality events remain unconfirmed due partly to limited health assessment tools. Mussel-monitoring activities often rely on population-level measurements, such as abundance and age structure, which reflect delayed responses to environmental conditions. Measures of organismal health would enable preemptive detection of declining condition before population-level effects manifest. Metabolomic analysis can identify shifts in biochemical pathways in response to stressors and changing environmental conditions; however, interpretation of the results requires information on inherent variability of metabolite concentrations in mussel populations. We targeted metabolites in the haemolymph of two common mussels, Lampsilis cardium and Lampsilis siliquoidea, from three Indiana streams (USA) using ultra-high-performance liquid chromatography combined with quadrupole time-of-flight mass spectroscopy. The influence of species, stream and sex on metabolite variability was examined with distance-based redundancy analysis. Metabolite variability was most influenced by species, followed by site and sex. Inter- and intraspecies metabolite variability among sexes was less distinct than differences among locations. We further categorized metabolites by occurrence and variability in mussel populations. Metabolites with high occurrence (Categories 1 and 2) included those indicative of energy status (catabolism versus anabolism; arginine, proline, carnitine, nicotinic acid, pantothenic acid), oxidative stress (proline, glutamine, glutamate) and protein metabolism (thymidine, cytidine, inosine). Metabolites with lower occurrence (Category 3) are constituents of assorted metabolic pathways and can be important biomarkers with additional temporal sampling to characterize their variability. These data provide a reference for future temporal (before/after) monitoring and for studies of stressor-metabolite linkages in freshwater mussels.

The Dose-Response Effect of Docosahexaenoic Acid on the Omega-3 Index in American Football Athletes.

PURPOSE: American-style football (ASF) players are at increased risk for head injuries and cardiovascular disease. n-3 polyunsaturated fatty acids are cardioprotective, and emerging evidence suggests benefits for protection against head injuries. However, fundamental knowledge of n-3 polyunsaturated fatty acid dosing in athletes such as ASF players remains poorly understood. Therefore, this study investigated the dose-response effect of docosahexaenoic acid (DHA) supplementation in red blood cells (RBC) and as the Omega-3 Index (O3I), in collegiate ASF players throughout a competitive season. METHODS: Sixty-nine ASF players were randomly assigned placebo (corn oil), or 2, 4, or 6 g·d -1 of DHA supplement. Blood samples were collected at eight time points (T1-T8) over 27 wk. RBC were extracted and analyzed by gas-liquid chromatography. Compliant players who had samples collected at all time points were analyzed. A repeated-measures ANOVA was conducted to assess the dose-response effect of DHA over time, and between-group differences at individual time points were assessed by one-way ANOVA followed by Tukey post hoc test. RESULTS: A significant dose and time interaction was found, and all supplement groups had significantly greater DHA in RBC compared with placebo from T2-T8 ( P < 0.05). Athletes receiving 6 g·d -1 of DHA had the greatest O3I, relative to other groups, and the O3I reached steady state by 15 wk. The 6 g·d -1 group surpassed >8% on the O3I at approximately twice the rate of the 4 g·d -1 group (8 vs 15 wk). CONCLUSIONS: Our findings provide important fundamental knowledge demonstrating a dose-response incorporation of DHA into RBC membranes up to 6 g·d -1 . Furthermore, 6 g·d -1 of DHA can be used to rapidly achieve a desired O3I (>8%) in athletes in only 8 wk.