Regulation of macronutrient metabolism by the gastrointestinal tract of ruminants

We set out to test the hypothesis that the gastrointestinal tract (GIT) of ruminant animals catabolizes amino acids (AAs) preferentially. We sought to determine whether this catabolism represents an obligate requirement, and whether this requirement stems from the need to generate energy or support other metabolic demands. The aim was to determine the composition of macronutrients (AAs, short chain fatty acids, and glucose) utilized by the GIT, and the influence of general and specific nutrient supplies on their routes of metabolism. Increasing protein supply to the small intestine did not alter the total amount of glucose removed by the GIT indicating, that glucose removal and therefore utilization is obligatory. In contrast, the net removal of AAs occurred at a constant proportion of arterial and luminal supplies. This translated to larger amounts of AAs removed from blood circulation, and from the lumen of the small intestine in response to increased small intestinal and blood supplies. In this respect, the net absorption of branched chain AAs was, unlike other essential AAs lower than 100%. Further, glutamate and glutamine net appearance across the whole GIT and small intestine was unaffected by protein supply. The disproportionate utilization of BCAA, glutamate, and glutamine as compared to other AAs suggested that their metabolism occurred toward specific metabolic requirements, possibly energy production. When Krebs cycle metabolism was investigated using individual AAs, glucose, and short chain fatty acids, leucine and valine did not contribute to the flux of Krebs cycle intermediates. Conversely, α-ketoglutarate flux originated mainly from glutamate, and to a lesser extent from glutamine. Though glucose was metabolized to pyruvate and lactate, glucose did not contribute to Krebs cycle intermediates. Overall, these results indicated that glutamate plays an important role in energy metabolism, and in insuring replenishment of Krebs cycle intermediates that leave the cycle via cataplerosis. Yet, the results raised new questions that ought to be addressed in future studies. The fate of glutamine carbon, the metabolic significance of leucine and valine deamination, and the role of glucose partial catabolism to lactate need to be investigated

Heat stress decreases metabolic flexibility in skeletal muscle of growing pigs

Heat-stressed pigs experience metabolic alterations, including altered insulin profiles, reduced lipid mobilization, and compromised intestinal integrity. This is bioenergetically distinct from thermal neutral pigs on a similar nutritional plane. To delineate differences in substrate preferences between direct and indirect (via reduced feed intake) heat stress effects, skeletal muscle fuel metabolism was assessed. Pigs (35.3 ± 0.8 kg) were randomly assigned to three treatments: thermal neutral fed ad libitum (TN, 21°C, n=8), heat stress fed ad libitum (HS, 35°C, n=8), and TN, pair-fed to HS intake (PF, n=8) for 7 days. Body temperature (TB) and feed intake (FI) were recorded daily. Longissimus dorsi muscle was biopsied for metabolic assays on days -2, 3, and 7 relative to initiation of environmental treatments. Heat stress increased TB and decreased FI (P \u3c 0.05). Heat stress inhibited incomplete fatty acid oxidation (P \u3c 0.05) and did not alter glucose oxidation. Metabolic flexibility decreased in HS pigs compared to TN and PF controls (P \u3c 0.05). Both phosphofructokinase and pyruvate dehydrogenase (PDH) activities increased in PF (P \u3c 0.05), however, TN and HS did not differ. Heat stress inhibited citrate synthase and beta hydroxyacyl-CoA dehydrogenase (βHAD) activities (P \u3c 0.05). Heat stress did not alter PDH phosphorylation or carnitine palmitoyltransferase 1 abundance, but reduced acetyl-CoA carboxylase 1(ACC1) protein abundance (P \u3c 0.05). In conclusion, HS decreased skeletal muscle fatty acid oxidation and metabolic flexibility, likely involving βHAD and ACC regulation

Protein synthesis in skeletal muscle of neonatal pigs is enhanced by administration of β-hydroxy-β-methylbutyrate.

Many low-birth-weight infants experience failure to thrive. The amino acid leucine stimulates protein synthesis in skeletal muscle of the neonate, but less is known about the effects of the leucine metabolite β-hydroxy-β-methylbutyrate (HMB). To determine the effects of HMB on protein synthesis and the regulation of translation initiation and degradation pathways, overnight-fasted neonatal pigs were infused with HMB at 0, 20, 100, or 400 μmol·kg body wt(-1)·h(-1) for 1 h (HMB 0, HMB 20, HMB 100, or HMB 400). Plasma HMB concentrations increased with infusion and were 10, 98, 316, and 1,400 nmol/ml in the HMB 0, HMB 20, HMB 100, and HMB 400 pigs. Protein synthesis rates in the longissimus dorsi (LD), gastrocnemius, soleus, and diaphragm muscles, lung, and spleen were greater in HMB 20 than in HMB 0, and in the LD were greater in HMB 100 than in HMB 0. HMB 400 had no effect on protein synthesis. Eukaryotic initiation factor (eIF)4E·eIF4G complex formation and ribosomal protein S6 kinase-1 and 4E-binding protein-1 phosphorylation increased in LD, gastrocnemius, and soleus muscles with HMB 20 and HMB 100 and in diaphragm with HMB 20. Phosphorylation of eIF2α and elongation factor 2 and expression of system A transporter (SNAT2), system L transporter (LAT1), muscle RING finger 1 protein (MuRF1), muscle atrophy F-box (atrogin-1), and microtubule-associated protein light chain 3 (LC3-II) were unchanged. Results suggest that supplemental HMB enhances protein synthesis in skeletal muscle of neonates by stimulating translation initiation

Leucine pulses enhance skeletal muscle protein synthesis during continuous feeding in neonatal pigs.

Infants unable to maintain oral feeding can be nourished by orogastric tube. We have shown that orogastric continuous feeding restricts muscle protein synthesis compared with intermittent bolus feeding in neonatal pigs. To determine whether leucine infusion can be used to enhance protein synthesis during continuous feeding, neonatal piglets received the same amount of formula enterally by orogastric tube for 25.25 h continuously (CON) with or without LEU or intermittently by bolus every 4 h (BOL). For the CON+LEU group, leucine pulses were administered parenterally (800 μmol·kg(-1)·h(-1)) every 4 h. Insulin and glucose concentrations increased after the BOL meal and were unchanged in groups fed continuously. LEU infusion during CON feeding increased plasma leucine after the leucine pulse and decreased essential amino acids compared with CON feeding. Protein synthesis in longissimus dorsi (LD), gastrocnemius, and soleus muscles, but not liver or heart, were greater in CON+LEU and BOL than in the CON group. BOL feeding increased protein synthesis in the small intestine. Muscle S6K1 and 4E-BP1 phosphorylation and active eIF4E·eIF4G complex formation were higher in CON+LEU and BOL than in CON but AMPKα, eIF2α, and eEF2 phosphorylation were unchanged. LC3-II-to-total LC3 ratio was lower in CON+LEU and BOL than in CON, but there were no differences in atrogin-1 and MuRF-1 abundance and FoxO3 phosphorylation. In conclusion, administration of leucine pulses during continuous orogastric feeding in neonates increases muscle protein synthesis by stimulating translation initiation and may reduce protein degradation via the autophagy-lysosome, but not the ubiquitin-proteasome pathway

Heat stress decreases metabolic flexibility in skeletal muscle of growing pigs

Heat-stressed pigs experience metabolic alterations, including altered insulin profiles, reduced lipid mobilization, and compromised intestinal integrity. This is bioenergetically distinct from thermal neutral pigs on a similar nutritional plane. To delineate differences in substrate preferences between direct and indirect (via reduced feed intake) heat stress effects, skeletal muscle fuel metabolism was assessed. Pigs (35.3 ± 0.8 kg) were randomly assigned to three treatments: thermal neutral fed ad libitum (TN, 21°C, n=8), heat stress fed ad libitum (HS, 35°C, n=8), and TN, pair-fed to HS intake (PF, n=8) for 7 days. Body temperature (TB) and feed intake (FI) were recorded daily. Longissimus dorsi muscle was biopsied for metabolic assays on days -2, 3, and 7 relative to initiation of environmental treatments. Heat stress increased TB and decreased FI (P This is a manuscript of an article published as Zhao, Lidan, Ryan P. McMillan, Guohao Xie, Samantha GLW Giridhar, Lance H. Baumgard, Samer El-Kadi, Joshua Selsby et al. "Heat stress decreases metabolic flexibility in skeletal muscle of growing pigs." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 315, no. 6 (2018): R1096-R1106. doi: 10.1152/ajpregu.00404.2017. Posted with permission.</p