Whole Body Rate Of Appearance Research Articles

Milk protein yield and mammary metabolism are affected by phenylalanine deficiency but not by threonine or tryptophan deficiency

Efficient milk protein synthesis requires that the essential AA be presented to the mammary gland in the right amount and proportion to maximize protein synthesis and minimize losses. This study investigated the effects of individual AA deficiencies on cow productivity, mammary metabolism, and glucose whole-body rate of appearance. Five Holstein cows were used in a 5×5 Latin square design trial with 10-d periods. Treatments were abomasal infusions of (1) water (CTL); (2) complete AA mixture (TAA); (3) TAA without Phe (No-Phe); (4) TAA without Thr (No-Thr); and (5) TAA without Trp (No-Trp). Each treatment was compared with TAA. Treatment did not affect milk, fat, or lactose yields. Arterial concentrations of Phe, Thr, and Trp decreased with their respective deletions by 60, 76, and 69%. In response to the decreased arterial supply of the deleted AA, mammary plasma flow significantly increased by 55% with No-Thr but did not increase with No-Phe or No-Trp. Mammary uptake of Phe was reduced by No-Phe, accompanied by a reduced milk protein yield; uptakes of Thr and Trp were not affected by their respective deletions, and milk protein yield did not decrease with these treatments. Deletion of Phe tended to reduce its mammary uptake relative to milk output (U:O), accompanied by an increased U:O of Tyr, but deletion of Thr and Trp did not affect the U:O of the corresponding AA. Plasma urea-N concentration was lower with CTL and tended to be higher with No-Phe. Arterial concentrations and mammary uptake of acetate, β-hydroxybutyrate, glucose, and lactate were unaffected by treatment. Treatment had no effect on glucose rate of appearance at the whole-body level. Lactose output as a percentage of glucose whole-body rate of appearance was not affected by treatment. Overall, the study indicated that a deficiency of Phe negatively affected productivity and mammary metabolism but that a deficiency of Thr or Trp did not.

Abomasal amino acid infusion in postpartum dairy cows: Effect on whole-body, splanchnic, and mammary glucose metabolism

Nine Holstein cows fitted with rumen cannulas and indwelling catheters in splanchnic blood vessels were used to study the effects of supplementing AA on milk lactose secretion, whole-body rate of appearance (WB-Ra) of glucose, and tissue metabolism of glucose, lactate, glycerol, and β-OH-butyrate (BHBA) in postpartum dairy cows according to a generalized randomized incomplete block design with repeated measures in time. At calving, cows were blocked according to parity (second and third or greater) and were allocated to 2 treatments: abomasal infusion of water (n=4) or abomasal infusion of free AA with casein profile (AA-CN; n=5) in addition to the same basal diet. The AA-CN infusion started with half the maximal dose at 1 d in milk (DIM) and then steadily decreased from 791 to 226g/d from DIM 2 to 29 to cover the estimated essential AA deficit. On DIM 5, 15, and 29, d[6,6-2H2]-glucose (23.7mmol/h) was infused into a jugular vein for 5h, and 6 blood samples were taken from arterial, portal, hepatic, and mammary sources at 45-min intervals, starting 1h after the initiation of the d[6,6-2H2]glucose infusion. Trans-organ fluxes were calculated as veno-arterial differences times plasma flow (splanchnic: downstream dilution of deacetylated para-aminohippurate; mammary: Fick principle using Phe+Tyr). Energy-corrected milk and lactose yields increased on average with AA-CN by 6.4kg/d and 353g/d, respectively, with no DIM × treatment interaction. Despite increased AA supply and increased demand for lactose secretion with AA-CN, net hepatic release of glucose remained unchanged, but WB-Ra of glucose tended to increase with AA-CN. Portal true flux of glucose increased with AA-CN and represented, on average, 17% of WB-Ra. Splanchnic true flux of glucose was unaltered by treatments and was numerically equivalent to WB-Ra, averaging 729 and 741mmol/h, respectively. Mammary glucose utilization increased with AA-CN infusion, averaging 78% of WB-Ra, and increased gradually as lactation advanced. Net portal, hepatic, splanchnic, and mammary fluxes of lactate, glycerol, and BHBA were not affected by AA infusion. Increasing the supply of AA in postpartum dairy cows elevated the WB-Ra of glucose without affecting the true liver glucose release. The greater WB-Ra of glucose with abomasal AA infusion seemed to originate mainly from greater true portal-drained viscera release of glucose. Glucose utilization by the portal-drained viscera was unaffected by abomasal AA infusion, but the exact mechanism behind the greater true portal glucose release could not be assessed in the current study. The increased mammary glucose uptake was in line with the increased milk lactose yield. In early postpartum lactation, the demand for AA seems to be so high that even with increased AA supply, cows have metabolic priorities for AA other than hepatic gluconeogenesis.

Effect of substitution of soybean meal by canola meal or distillers grains in dairy rations on amino acid and glucose availability

Canola meal (CM) or by-products of ethanol production (dried distillers grain, DDG) may offer an economical alternative to soybean meal (SBM) in North American dairy rations. These protein supplements can effectively replace SBM and, in 2 recent meta-analyses, CM had a positive effect on milk and milk protein yields compared with SBM. The objective of this study was to determine if the positive responses observed with inclusion of CM in dairy rations could be explained by an increased availability of His, Lys, Met, or glucose. Eight Holstein dairy cows were used in a replicated 4×4 Latin square with 14-d periods. Cows were fed isonitrogenous (17.2% crude protein) and isoenergetic (1.56Mcal/kg of net energy of lactation) diets formulated to slightly exceed nutrient requirements. Diets contained 38% grass hay and 62% corn-based concentrate including SBM, CM, corn high-protein DDG (HPDDG), or wheat DDG plus solubles (WDDGS) as the single protein supplement. The effect of protein supplements on availability of His, Lys, Met, and glucose was estimated using variations in the whole-body (WB) flux of these nutrients, determined by isotopic dilution. As planned, dry matter intake and milk and milk protein yields were not affected by treatments and averaged 23.7, 31.4, and 1.14kg/d, respectively. Lactose yield did not differ among diets although milk lactose content tended to be lower with CM and WDDGS diets than with SBM and HPDDG diets. Lysine availability was affected by treatments: the highest WB irreversible loss rate (ILR) was observed for the CM diet (371g/d) and the lowest for HPDDG diet (290g/d); values for SBM and WDDGS were intermediate (330 and 316g/d, respectively). Availability of His and Met did not vary among diets and WB ILR averaged, respectively, 129 and 124g/d; the CM diet, however, had numerically the highest His and Met ILR. Plasma concentrations of most of the essential AA were higher with the CM diet and lower with the HPDDG diet, the exception being Leu for which the concentration was highest for the HPDDG diet. Glucose WB rate of appearance was altered by diet, with the highest mean observed for SBM (3,036g/d) and the lowest for CM (2,795g/d); the 2 diets with the lowest WB glucose rate of appearance (CM and WDDGS) also had the lowest dietary starch concentration. Overall, this study suggested that positive responses in milk and milk protein yields observed with inclusion of CM in dairy rations could be linked to a greater supply of metabolizable protein, including some essential AA, especially His, Lys, and Met, as glucose availability was certainly not increased in cows fed the CM diet.

Adipose triglyceride lipase and hormone-sensitive lipase protein expression in subcutaneous adipose tissue is decreased after an isoenergetic low-fat high-complex carbohydrate diet in the metabolic syndrome

The objective was to determine the contribution of dietary fat quantity and composition to lipolysis and lipolytic gene expression in humans in relation to obesity, insulin resistance, and the metabolic syndrome (MetS). Men and women with the MetS were randomly assigned to one of four isoenergetic diets: a high-fat saturated fat diet (n=10), a high-fat monounsaturated fat diet (n=7), and two low-fat high-complex carbohydrate (LFHCC) diets, one supplemented with 1.24g/day long-chain n-3 PUFA (LFHCC: n=7, LFHCCn-3: n=8). Subcutaneous adipose tissue biopsies were taken before and after the 12-week dietary intervention period. ATGL and HSL mRNA and protein expression was determined. Whole body rate of appearance of free fatty acids (RaFFA) was determined by intravenous infusion of [2H2]-palmitate in a subgroup of men (n=20). Adipose tissue ATGL and HSL mRNA and protein expression was not affected by alterations in dietary fat composition. Pooled analysis comparing the low- and high-fat diets showed that ATGL and HSL protein expression was significantly reduced after the LFHCC diets (P=.04), irrespective of long-chain n-3 PUFA. Moreover, LFHCC diets lowered fasting insulin, HOMAIR, and (LDL)-cholesterol concentrations (P≤.05). Changes in ATGL and HSL protein expression was positively associated with changes in whole body RaFFA (P<.03). The low-fat high-complex carbohydrate diets reduced ATGL and HSL protein expression and significantly improved circulating lipids and insulin sensitivity. Under isoenergetic conditions, dietary fat quantity, rather than composition, may be most important for modulating subcutaneous adipose tissue ATGL and HSL protein expression.

Nitric oxide and l-arginine metabolism in a devascularized porcine model of acute liver failure

In acute liver failure (ALF), the hyperdynamic circulation is believed to be the result of overproduction of nitric oxide (NO) in the splanchnic circulation. However, it has been suggested that arginine concentrations (the substrate for NO) are believed to be decreased, limiting substrate availability for NO production. To characterize the metabolic fate of arginine in early-phase ALF, we systematically assessed its interorgan transport and metabolism and measured the endogenous NO synthase inhibitor asymmetric dimethylarginine (ADMA) in a porcine model of ALF. Female adult pigs (23-30 kg) were randomized to sham (N = 8) or hepatic devascularization ALF (N = 8) procedure for 6 h. We measured plasma arginine, citrulline, ornithine levels; arginase activity, NO, and ADMA. Whole body metabolic rates and interorgan flux measurements were calculated using stable isotope-labeled amino acids. Plasma arginine decreased >85% of the basal level at t = 6 h (P < 0.001), whereas citrulline and ornithine progressively increased in ALF (P < 0.001 and P < 0.001, vs. sham respectively). No difference was found between the groups in the whole body rate of appearance of arginine or NO. However, ALF showed a significant increase in de novo arginine synthesis (P < 0.05). Interorgan data showed citrulline net intestinal production and renal consumption that was related to net renal production of arginine and ornithine. Both plasma arginase activity and plasma ADMA levels significantly increased in ALF (P < 0.001). In this model of early-phase ALF, arginine deficiency or higher ADMA levels do not limit whole body NO production. Arginine deficiency is caused by arginase-related arginine clearance in which arginine production is stimulated de novo.

Effect of amino acid or casein supply on whole-body, splanchnic, and mammary glucose kinetics in lactating dairy cows

This study was conducted to establish how AA supplied in a free form or as protein (casein, CN) affect the whole-body rate of appearance (WB Ra) of glucose, splanchnic and mammary glucose kinetics, and milk lactose secretion in lactating dairy cows. Five Holstein cows fitted with a rumen cannula and permanent indwelling catheters in the abomasum, portal, hepatic, and mesenteric veins, and one mesenteric artery, were used in a Youden square with 4 periods of 14 d each. Cows were fed a hay-based diet providing 100 and 70% of their net energy and metabolizable protein (MP) requirements, respectively. Treatments consisted of abomasal infusions of water (70% of MP requirements: control, Con), free AA (95% of MP requirements: AA1; and 120% of MP requirements: AA2), or CN (95% of MP requirements: CN1). The free AA mixture had the same profile as CN. On d 14 of each period, [6,6-2H2]glucose (25.8 mmol/h) was infused into a jugular vein, and blood samples (n=8) were taken over 4h from arterial, portal, hepatic, and mammary sources to measure glucose enrichment and concentration. Splanchnic and mammary plasma flows were determined by downstream dilution of para-aminohippurate and with the Fick principle, respectively. The last 6 milkings of each period were weighed and sampled to measure the yields of milk and components. The AA1 and CN1 treatments were not different for any of the measured parameters. Supplying AA linearly increased glucose WB Ra (AA2 vs. Con: +151 mmol/h) and liver net flux (+149 mmol/h). Utilization of glucose from the plasma compartment by the portal-drained viscera and liver and true portal absorption were not affected by AA supply. From these observations, we suggest that the increased WB Ra was due to increased net hepatic production. The AA from the infusion, in excess of that used to cover the increase in milk protein, were converted to glucose with an apparent efficiency close to 100% of maximum theoretical efficiency. Milk and lactose yields increased linearly with infusions of AA, by 14 and 16% with AA2 treatment, respectively. However, mammary glucose uptake was not significantly altered by AA infusions; this suggests that the mammary gland exerts active control on the uptake and utilization of glucose. For all treatments, the sum of true portal glucose absorption and true hepatic glucose production contributed more than 99% of WB Ra in the lactating cow; this would suggest that renal glucose synthesis makes only a small contribution to WB Ra under these conditions.

Is glutamine still an important precursor of citrulline?

Is glutamine still an important precursor of citrulline?

Methionine metabolism in human pregnancy

Hyperhomocysteinemia during pregnancy, which is a consequence of perturbations in methionine and/or folate metabolism, has been implicated in adverse outcomes such as neural tube defects, preeclampsia, spontaneous abortion, and premature delivery. The adaptive changes in methionine metabolism during pregnancy in humans have not been determined. Our objective was to examine the kinetics of methionine and its rate of transsulfuration and transmethylation in healthy women with advancing gestation. The whole-body rate of appearance (Ra) of methionine and phenylalanine was measured in healthy pregnant women during the first (n = 10), second (n = 5), and third (n = 10) trimesters of pregnancy. These data were compared with those for nonpregnant women (n = 8). Tracers [1-(13)C]methionine, [C(2)H(3)]methionine, and [(2)H(5)]phenylalanine were administered as prime-constant rate infusions. The effect of enteral high-protein, mixed-nutrient load on tracer-determined variables was also examined. In pregnant women, the Ra of phenylalanine was significantly (P < 0.05) lower in the first trimester than in the second and third trimesters and was significantly lower than that in nonpregnant women. A linear positive correlation was evident between gestational age and phenylalanine Ra. The fractional rate and total rate of transsulfuration of methionine was significantly (P < 0.05) higher during the first trimester, whereas the rate of transmethylation was higher during the third trimester. Plasma concentrations of total cysteine and homocysteine were lower during pregnancy. Uncomplicated pregnancy in humans is associated with a higher rate of transsulfuration early in gestation and a higher rate of transmethylation of methionine in late gestation. These data may have implications for understanding the role of methionine and homocysteine in complications of pregnancy and for the nutritional care of pregnant women.

Skeletal Muscle Protein Anabolic Response to Increased Energy and Insulin Is Preserved in Poorly Controlled Type 2 Diabetes

Type 2 diabetes (T2DM) subjects failing diet treatment are characterized by hyperinsulinemia and insulin resistance leading to fasting and postprandial hyperglycemia and hyperlipidemia. Energy is essential for allowing the process of protein synthesis to proceed. Additionally, insulin can stimulate protein synthesis in human muscle. The aims of this study were to determine if poorly controlled T2DM affects postabsorptive muscle protein anabolism, and if the muscle anabolic response to hyperinsulinemia with high energy availability is maintained. Control (n = 6) and T2DM subjects (n = 6) were studied in the postabsorptive state and during an isoenergetic high nutritional energy clamp (relative to postabsorptive state). Muscle protein synthesis and breakdown (nmol . min(-1) . 100 g leg muscle(-1)) were assessed using stable isotope methodology, femoral arterio-venous sampling, muscle biopsies, and a three-pool model to calculate protein turnover. Postabsorptive phenylalanine net balance and whole body rate of appearance (Ra) were not different between groups; however, basal muscle protein breakdown was higher in T2DM (94 +/- 9) than in controls (58 +/- 12) (P < 0.05) and muscle protein synthesis tended (P = 0.07) to be elevated in T2DM (66 +/- 14) compared with controls (39 +/- 6). During the clamp, net balance increased, whole body Ra and muscle protein breakdown decreased (P < 0.05), and muscle protein synthesis tended to decrease (P = 0.08) to a similar extent in both groups. We conclude that postabsorptive muscle protein turnover is elevated in poorly controlled T2DM, however, there is no excessive loss of muscle protein because net balance is not different from controls. Moreover, the anabolic response to increased insulin and energy availability is maintained in T2DM.

Decreased circulating arginine in ALF mice does not compromise whole body NO production

In fulminant hepatic encephalopathy (HE), the plasma level of the precursor of nitric oxide (NO) Arginine is reduced. In contrast, nitric oxide (NO) production, measured as plasma NOx is increased. This contradiction could be due to failure of plasma NOx to represent NO production. Therefore, we studied whole body (Wb) ARG (precursor of NO) and NO metabolism using stable isotope technology in a newly developed acetaminophen (APAP) induced acute liver failure (ALF) mouse model with pre-coma HE after 20h. Method: In post absorptive C57Bl6/J mice, an oral dose of 200+200 mg/kg of APAP or drinking water (control) was given at t=0+1 h. At T=20 h, Wb ARG and Arginine to NO and Urea/Ornithine conversion was measured in carotid artery/jugular vein catherized mice under anaesthesia using a primed-continuous infusion of 15N-CIT, 15N2-2H2-ARG and 13C-UREA stable isotopes. At plasma steady state, arterial (art) plasma was collected. Whole body rate of appearance (WbRa), NO production, de novo ARG and rate of urea converted from ARG (ARG-UREA) were calculated. Statistics: t-test, sign p<0.05, data as mean ± SEM. Results Twelve of the 17 APAP animals had ALT>1000 IU/l (ALF group) and showed behaviour changes at a level of pre coma. Art ARG was decreased in the ALF group (65 ± 15 vs. 134 ± 6 μM). Art ORN (190 ± 19 vs. 84 ± 7 μM) and ARG-UREA conversion (19 ± 3 vs. 13 ± 1 nmol/10 gr bw/min) were increased. WbRa of ARG, Wb NO production and de novo ARG were unchanged compared to control animals. Conclusion Reduced plasma ARG is probably caused by release of the enzyme arginase from the injured liver and subsequent ARG to ORN/UREA conversion. Reduced Arginine availablity, however, did not affect NO production at this early stage of HE.

Gender differences in basal protein kinetics in young adults.

Gender affects energy expenditure and influences the relative utilization of carbohydrate and fat as fuels. However, little is known about the possible effects of gender on protein metabolism. Thus, we compared whole body and plasma (albumin and fibrinogen) protein kinetics in the basal postabsorptive state in young, untrained volunteers divided into two groups according to gender (women: n=17; age, 24+/-4 yr; men: n=17; age, 25+/-2 yr). The two groups were matched for body mass index. Protein kinetics were measured by means of L-[1-14C]leucine infusion. The leucine whole body rate of appearance, an index of proteolysis, and nonoxidative rate of disappearance, an index of protein synthesis, were similar in the two groups. However, the leucine oxidation rate was significantly lower in women compared to men (0.23+/-0.07 vs. 0.31+/-0.08 micromol/kg min; P=0.0062). Similar results were obtained when data were adjusted for estimated body composition. Albumin and fibrinogen fractional secretion rates were not different in the two groups. In conclusion, in the basal state leucine oxidation is lower in women than in men regardless of body composition. This could be one of the factors contributing to the lower metabolic rate in women.

Fasting and postprandial phenylalanine and leucine kinetics in liver cirrhosis.

To investigate body protein turnover and the pathogenesis of increased concentration of plasma phenylalanine in liver cirrhosis, we have studied phenylalanine and leucine kinetics in cirrhotic (diabetic and nondiabetic) patients, and in normal subjects, both in the postabsorptive state and during a mixed meal, using combined intravenous and oral isotope infusions. Postabsorptive phenylalanine concentration and whole body rate of appearance (Ra) were approximately 40% greater (P < 0.05) in patients than in controls. Leucine concentrations were comparable, but intracellular leucine Ra was also increased (P < 0.05), suggesting increased whole body protein breakdown. Postprandial phenylalanine Ra was also greater (P < 0.05) in the patients. This difference was due to a diminished fractional splanchnic uptake of the dietary phenylalanine (approximately 40% lower in the cirrhotics vs. controls, P < or = 0.05). Postprandial leucine Ra was also increased in the patients, but splanchnic uptake of dietary leucine was normal. Thus both increased body protein breakdown and decreased splanchnic extraction of dietary phenylalanine can account for the increased phenylalanine concentrations in liver cirrhosis.

Isotopic estimation of CO2 production during exercise before and after endurance training

Endurance training reduces the rate of CO2 release (i.e., VCO2) during submaximal exercise, which has been interpreted to indicate a reduction in carbohydrate oxidation. However, decreased ventilation, decreased buffering of lactate, and/or increased fixation of CO2 could also account for a lower VCO2 after training. We therefore used a primed continuous infusion of NaH13CO3 to determine the whole body rate of appearance of CO2 (RaCO2) in seven men during 2 h of cycle ergometer exercise at 60% of pretraining peak O2 uptake (VO2peak) before and after endurance training. RaCO2 is independent of the above-described factors affecting VCO2 but may overestimate net CO2 production due to pyruvate carboxylation and subsequent isotopic exchange in the tricarboxylic acid cycle. Training consisted of cycling at 75-100% VO2peak for 45-90 min/day, 6 days/wk, for 12 wk and increased VO2peak by 28% (P < 0.001). VCO2 during submaximal exercise was reduced from 86.8 +/- 3.7 to 76.2 +/- 4.2 mmol/min, whereas RaCO2 fell from 88.9 +/- 4.0 to 76.4 +/- 4.4 mmol/min (both P < 0.001). VCO2 and RaCO2 were highly correlated in the untrained (r = 0.98, P < 0.001) and trained (r = 0.99, P < 0.001) states, as were individual changes in VCO2 and RaCO2 with training (r = 0.88, P < 0.01). These results support the hypothesis that endurance training decreases CO2 production during exercise. The magnitude and direction of this change cannot be explained by reported training-induced alterations in amino acid oxidation, indicating that it must be the result of a decrease in carbohydrate oxidation and an increase in fat oxidation.(ABSTRACT TRUNCATED AT 250 WORDS)