Response of glucose disposal to hyperinsulinaemia in human hypothyroidism and hyperthyroidism.

We have examined insulin action on glucose metabolism in six hypothyroid patients before and after regular thyroid hormone treatment, and in six healthy volunteers before and after transient induction of moderate hyperthyroidism. Insulin was infused under euglycaemic and eukalaemic clamps. An appropriate amino acid infusion was used to blunt insulin-induced decreases in amino acid levels. Glucose kinetics were assessed using a primed continuous infusion of [6,6-(2)H(2)]glucose. The results showed that basal plasma insulin and glucose levels (i.e. before infusion) were similar in each case. Despite similar insulin infusion rates, the plateau value of insulin was lower after thyroid treatment in both hypothyroid patients and healthy volunteers. The rate of exogenous glucose needed to maintain plasma glucose at a steady-state level was increased by thyroid hormone in hypothyroid patients (P <0.05), but not in healthy volunteers. Thyroid treatment resulted in a significant increase in basal glucose disposal in both groups (P <0.05). Insulin, in conjunction with glucose and amino acids, significantly stimulated glucose disposal (P <0.05) under all conditions. The incremental increase in glucose disposal after infusion tended to be higher following thyroid hormone treatment, but this was not statistically significant. However, the ratio of the incremental increase in glucose disposal to the increase in plasma insulin was significantly improved after thyroid hormone treatment in hypothyroid patients (P <0.05). It was also increased in healthy volunteers, but not significantly. We conclude that thyroid hormones improve the ability of insulin to stimulate glucose disposal related to insulinaemia. This phenomenon may be highly sensitive, because it was only apparent at low thyroid hormone levels.

Response of leucine metabolism to hyperinsulinemia in hypothyroid patients before and after thyroxine replacement.

We have investigated the effect of hypothyroidism and insulin on protein metabolism in humans. Six hypothyroid patients were studied in a postabsorptive state before and after 5 months of regular treatment for hypothyroidism (153 +/- 17 microg/day of L-T4). The effect of insulin was assessed under hyperinsulinemic euglycemic and eukalemic conditions. Insulin was infused for 140 min at 0.0063 +/- 0.0002 nmol/kg x min. An amino acid infusion was used to blunt insulin-induced hypoaminoacidemia. Whole body protein turnover was measured using L-[1-13C] leucine. When compared to L-T4-induced subclinical thyrotoxic state, hypothyroidism induced a significant decrease (P < 0.05) in leucine endogenous appearance rate (a reflection of proteolysis; 0.89 +/- 0.09 vs. 1.33 +/- 0.05 micromol/kg x min), oxidation (0.19 +/- 0.02 vs. 0.25 +/- 0.03 micromol/kg x min), and nonoxidative disposal (a reflection of protein synthesis; 0.87 +/- 0.11 vs. 1.30 +/- 0.05 micromol/ kg x min). Insulin lowered proteolysis during both the subclinical thyrotoxic and hypothyroid states. Hypothyroidism impaired the antiproteolytic effects of insulin. Thyroid hormones are, therefore, essential for the normal antiproteolytic action of insulin.

Threonine and methionine are limiting amino acids for protein synthesis in patients with AIDS.

This study was conducted to identify the most rate-limiting amino acids for whole-body protein synthesis in acquired immunodeficiency syndrome (AIDS) patients. We postulated that an essential amino acid that would be rate limiting in AIDS should have a low basal plasma concentration and should remain at a low level during amino acid infusion. Seven male AIDS patients (median age 37 y, CD4 cell count: 76 mm-3) without any clinically active opportunistic infection during the month before the experiment were infused intravenously with a complete amino acid-glucose mixture for 2.5 h. Eight healthy volunteers were used as controls. Before the infusion, the concentrations of most free essential amino acids (methionine, threonine, histidine, isoleucine, leucine and tryptophan) were significantly lower (P < 0.05) in AIDS patients than in controls. Most plasma free essential amino acids increased significantly during infusion. However, the absolute increase above basal levels for threonine, valine, lysine, (P < 0.05) and methionine (P < 0.073) was smaller in AIDS patients than in control subjects. Thus, threonine and possibly methionine may be rate limiting for whole-body protein synthesis in AIDS patients, suggesting that there are selective amino acid requirements in patients with AIDS.

Euglycemic hyperinsulinemia and hyperaminoacidemia decrease skeletal muscle ubiquitin mRNA in goats.

Insulin inhibits protein breakdown at the whole body level, but neither the tissues nor the proteolytic pathways on which insulin exerts its antiproteolytic effect are well characterized. We measured the effects of insulin on mRNA levels for cathepsin D and m-calpain (a lysosomal and Ca2(+)-dependent proteinase, respectively) and ubiquitin (a component of ubiquitin-dependent proteolysis) in skeletal muscle, skin, liver, and intestine. We used a 6-h hyperinsulinemic, euglycemic, and hyperaminoacidemic clamp in goats, a species in which insulin markedly inhibited whole body protein breakdown under similar conditions [S. Tesseraud, J. Grizard, E. Debras, I. Papet, Y. Bonnet, G. Bayle, and C. Champredon. Am. J. Physiol. 265 (Endocrinol. Metab. 28): E402-E413, 1993]. Hyperinsulinemia and hyperaminoacidemia had no effect on cathepsin D, m-calpain, and ubiquitin mRNA levels in liver, skin, and jejunum. In contrast, depressed ubiquitin mRNA levels were seen in skeletal muscle without any concomitant reduction in mRNA levels for cathepsin D, m-calpain, and other components of the ubiquitin-dependent proteolytic pathway. The reduced ubiquitin mRNA levels in skeletal muscle may represent a possible mechanism explaining the antiproteolytic effect of insulin in vivo.

Response of leucine metabolism to hyperinsulinemia under amino acid replacement in experimental hyperthyroidism.

We investigated the responsiveness of protein metabolism to insulin as a mediator of the protein catabolic response to hyperthyroidism in humans. Six healthy volunteers were studied in a postabsorptive state before and after oral intake of thyroid hormones (2 micrograms.kg-1.day-1 L-thyroxine for 6 wk along with 1 microgram.kg-1.day-1 triiodothyronine for the last 2 wk). Insulin was infused at 7.14 nmol.kg-1.min-1 for 140 min under euglycemic and eukalemic clamps. An appropriate amino acid infusion was used to blunt insulin-induced hypoaminoacidemia. Leucine kinetics were assessed using a primed continuous infusion of L-[1-13C]leucine. Hyperthyroidism induced a significant increase (P < 0.05) in leucine endogenous appearance rate (a reflection of proteolysis; 2.15 +/- 0.06 vs. 1.76 +/- 0.03 mumol.kg-1.min-1 in the control state), oxidation (0.54 +/- 0.04 vs. 0.47 +/- 0.07), and nonoxidative disposal (a reflection of protein synthesis; 1.80 +/- 0.06 vs. 1.45 +/- 0.06). Insulin lowered proteolysis. Further hyperthyroidism improved the ability of insulin to inhibit proteolysis, whether considered as an absolute decrease (-0.57 +/- 0.02 vs. -0.45 +/- 0.05 mumol.kg-1.min-1, P < 0.05) or related to insulinemia [1.59 +/- 0.11 vs. 1.01 +/- 0.08 mumol leucine.kg-1.min-1/(nmol insulin/l), P < 0.05]. Insulin also moderately (but significantly P < 0.05) lowered protein synthesis in both control and hyperthyroid states. These changes in insulin action may provide a mechanism to save body protein during hyperthyroidism.

Effect of hyperinsulinemia and hyperaminoacidemia on muscle and liver protein synthesis in lactating goats.

The experiment was carried out to clarify the roles of insulin and amino acids on protein synthesis in fed lactating goats (30 days postpartum). Protein synthesis in the liver and various skeletal muscles was assessed after an intravenous injection of a large dose of unlabeled valine containing a tracer dose of L-[2,3,4-3H]valine. The animals were divided into three groups. Group I was infused with insulin (1.7 mumol/min) for 2.5 h under glucose, potassium, and amino acid replacement. Group A was infused with an amino acid mixture to create stable hyperaminoacidemia for 2.5 h. Group C animals were controls. The fractional synthesis rates (FSR) were 31.5 +/- 2.2, 6.5 +/- 0.4, 4.3 +/- 0.8, 4.0 +/- 1.2, 3.9 +/- 1.2, and 3.6 +/- 0.4%/day (SD) in liver, masseter, diaphragm, anconeus, semitendinosus, and longissimus dorsi, respectively, for group C. Neither hyperinsulinemia in group I nor hyperaminoacidemia in group A had not affected by hyperinsulinemia but was stimulated by hyperaminoacidemia (+30%, P < 0.05). In contrast to previous experiments in which a labeled amino acid was constantly infused, this study revealed a stimulating effect of amino acids on protein synthesis in the liver but not in skeletal muscles. As previously observed in studies with the constant-infusion method, insulin had no effect on protein synthesis.

Leucine metabolism in lactating and dry goats: effect of insulin and substrate availability.

Early lactating goats show insulin resistance with respect to extramammary glucose utilization. However, much less is known about the two major factors, insulin and plasma amino acid concentration, that regulate protein metabolism in lactating goats. To examine this question, the in vivo effect of acute insulin was studied in goats during early lactation (12-31 days postpartum), midlactation (98-143 days postpartum), and the dry period (approximately 1 yr postpartum). Insulin was infused (at 0.36 or 1.79 nmol/min) under euglycemic and eukaliemic clamps. In addition, appropriate amino acid infusion was used to blunt insulin-induced hypoaminoacidemia or to create hyperaminoacidemia and maintain this condition under insulin treatment. Leucine kinetics were assessed using a primed continuous infusion of L-[1-14C]-leucine, which started 2.5 h before insulin. In all animals the insulin treatments failed to stimulate the nonoxidative leucine disposal (an estimate of whole body protein synthesis) under both euaminoacidemic and hyperaminoacidemic conditions. Thus, in goat as well as humans, infusion of insulin fails to stimulate protein synthesis even when combined with a substantially increased provision of amino acids. In contrast, insulin treatments caused a dose-dependent inhibition of the endogenous leucine appearance (an estimate of whole body protein degradation). Under euaminoacidemia the initial slope from the plot of the endogenous leucine appearance as a function of plasma insulin (an insulin sensitivity index) was steeper during early lactation than when compared with the dry period. A similar trend occurred during midlactation but not to any significant degree. These differences were abolished under hyperaminoacidemia. It was concluded that the ability of physiological insulin to inhibit protein degradation was improved during lactation, demonstrating a clear-cut dissociation between the effects of insulin on protein and glucose metabolism. This adaptation no doubt may provide a mechanism to save body protein.

Effect of insulin in conjunction with glucose, amino acids and potassium on net metabolism of glucose and amino acids in the goat mammary gland.

The hyperinsulinaemic euglycaemic insulin clamp technique was used to study the effect of insulin on the arterio-venous concentration differences of glucose and amino acids across the mammary gland in dairy goats. Insulin was given in conjunction with K to prevent insulin hypokalaemia. Appropriate amino acid infusion was used to blunt insulin-induced hypoaminoacidaemia or to create hyperaminoacidaemia and maintain this state under insulin treatment. Hyperaminoacidaemia alone only stimulated mammary leucine uptake but did not significantly modify the net metabolism of other amino acids and glucose. Insulin infusion at physiological level in conjunction with glucose, KCl-NaCl and amino acids failed to alter mammary uptake of glucose and essential amino acids; occasional increase in arginine extraction and decrease in tyrosine extraction were exceptions. Thus these new experimental conditions did not reveal any galactopoietic effect of insulin.

Parathyroid hormone-related peptide might stimulate calcium secretion into the milk of goats.

Parathyroid hormone-related peptide (PTHrP) has been shown to stimulate the placental calcium pump in pregnant ewes. Recent studies also suggest a physiological role for this peptide during lactation. In the present work, we have studied the possible role of synthetic human PTHrP(1-34), (1-86) and (140-173) fragments on calcium, inorganic phosphorus and magnesium secretion into milk of four adult goats in early lactation. This was done by measuring differences in calcium, inorganic phosphorus and magnesium concentrations in the carotid artery and superficial epigastric vein, the concentration of these minerals in milk and milk production. Both PTHrP(1-34) and (1-86) fragments stimulated calcium, inorganic phosphorus and magnesium uptake by the mammary gland and secretion into milk, without any significant effect on milk production. Therefore PTHrP might have important physiological roles in the pregnant and/or lactating female, by regulating calcium transport through the placenta and mammary gland.

Methionine flux and tissue protein synthesis in lactating and dry goats.

Whole-body methionine flux (rate of irreversible loss from plasma) and tissue protein synthesis were estimated in dry and early lactating goats (10-14 d postpartum) by intravenous infusion of L-[35S]methionine. Tissue protein mass was significantly (p less than 0.05) higher for mammary gland and liver but lower for carcass in lactating animals. The plasma methionine flux was higher during lactation (8.5 vs. 5.1 g/d). The fractional synthesis rates of tissue proteins (Ksp: %/d) were lower during lactation for some muscles, especially the masseter muscle (1.46 vs. 2.15), and for skin (0.59 vs. 1.22) and the pooled head plus feet fraction (1.64 vs. 2.31), but the rates were greatly increased in mammary gland (42 vs. 3). The non-mammary methionine flux (plasma flux minus the flux corresponding to milk methionine output and methionine utilization for mammary protein synthesis) was significantly (p less than 0.05) lower for the lactating goats than for the dry group (93 vs. 131 mg.d-1.kg empty body weight-1). This is in agreement with the lower rates of protein synthesis in carcass (542 vs. 948 mg.d-1.kg empty body weight-1) and skin (93 vs. 189) for lactating compared to dry goats. It can be inferred from these data that in early lactation, when nutrient requirements of animals are not adequately met, an adaptative mechanism occurs that allows amino acids to be available for the mammary gland by a decrease of their utilization in some extramammary tissues.

Insulin sensitivity and responsiveness during lactation and dry period in goats.

To investigate the role of insulin in partitioning nutrients between the mammary gland and other tissues during lactation in ruminants, euglycemic-hyperinsulinemic clamps were performed in goats during early lactation (15-26 days postpartum), midlactation (78-91 days postpartum), and dry period (169-194 days postpartum). Insulin was infused at 0.4, 0.7, 1.9, 4.4, and 10 micrograms/min. Basal plasma glucose was constant during all periods despite the fact that basal glucose utilization was approximately 3 times higher during lactation than dry period. Basal plasma insulin was similar during early lactation and dry period but increased during midlactation. Insulin infusion resulted in a dose-dependent stimulation of glucose utilization. The insulin-stimulated glucose utilization above basal was greatly impaired during early lactation when compared with dry period, but this only occurred at very high plasma insulin. Insulin infusion also resulted in a decrease in glucose production; the maximal insulin effect is achieved at the lowest insulin infusion rate. The ability of insulin to decrease glucose production was significantly improved during early lactation when compared with dry period. This phenomenon may provide a mechanism to save gluconeogenic substrates during early lactation. In contrast, midlactation did not result in any significant change in insulin action with both glucose utilization and glucose production.

Metabolism and action of insulin and glucagon in goat during lactating and dry period.

The metabolism and action of insulin and glucagon were investigated in goats during mid lactating (50 days postpartum) and during the dry period. The animals were fed hay and concentrate during lactation (1:1) and only hay during dry period. Pulse doses of unlabelled insulin and glucagon were injected intravenously. The disappearance of insulin from the circulation was faster during lactation than during dry period; the metabolic clearance rate of insulin was significantly increased during lactation. In contrast, the kinetic parameters of glucagon disappearance were very similar during the two periods. Basal plasma hormones (i.e. before hormone injection) were higher during lactation than during dry period; the molar ratio insulin:glucagon was left unchanged. The increase in plasma insulin following glucagon-stimulated hyperglycaemia was similar during the two periods. The ability of insulin to elicit a decrease in blood glucose was markedly impaired during lactation when compared to dry period. In contrast the ability of glucagon to increase blood glucose was slightly improved during lactation. Those endocrine changes could be related to the effect of both lactation and diet.

[Findings and hormonal coordination of protein metabolism in ruminants]. / Orientations et coordination hormonale du métabolisme protéique chez les ruminants.

Protein metabolism roughly exhibits the same characteristics in ruminants and non-ruminant species. Changes in whole-body protein mass are the result of the balance between the simultaneous protein synthesis and breakdown. Essential amino acids are either incorporated into protein or degraded. Insulin, growth hormone and glucagon have been shown to regulate those metabolic pathways in ruminants. Alteration of food intake both decreases protein synthesis and protein breakdown. Protein synthesis is affected to a greater extent than protein breakdown. So protein synthesis is the main factor controlling N balance in response to alteration of food intake. The decrease in protein synthesis may be related to an impairment of plasma insulin. Protein synthesis and breakdown both decrease throughout development. Protein deposition decreases throughout development because protein synthesis declines more rapidly than protein breakdown. The hormonal coordination of those changes depends on the age of the animals. For example the decrease in protein synthesis in the postnatal growth period may be due to either the decrease in plasma growth hormone or to the impairment in cellular insulin receptors. Due to milk protein synthesis in the mammary gland, lactating animals exhibit a large increase in whole-body protein synthesis. Surprisingly, protein synthesis decreases in some non-mammary tissues. This represents a mechanism for a greater partitioning of amino acids towards milk at the expense of body proteins. We recently demonstrated that insulin could be involved in that adaptative process.

[Digestion of milk proteins and metabolism of methionine in the newborn lamb]. / Digestion des protéines de lait et métabolisme de la méthionine chez l'agneau nouveau-né.

Methionine absorption and catabolism were studied in 4 newborn lambs during the first 8 h after birth. Lambs were hourly fed 50 ml goat milk labelled with 35S-methionine and 35S-cysteine. The free amino acid levels and the specific activity of free methionine were measured in jugular blood samples collected 1 h (just before the first meal), 4, 6 and 8 h after birth and in the portal blood 8 h after birth. Specific activities of protein-bound methionine and cysteine were measured in the milk and then in the abomasal and intestinal contents as well as in the liver, intestine and whole body proteins, 8 h after birth. The jugular blood levels of free valine, isoleucine, leucine, phenylalanine and histidine increased significantly between 1 and 8 h whereas the levels of free alanine, serine, glycine, citrulline and 3-methylhistidine decreased. The concentrations of most free amino acids were 30% higher in portal than in jugular blood. In the abomasal contents, the specific activities of methionine and cysteine were 96 and 168%, respectively of that of ingested milk and in the intestinal contents the corresponding values were 24 and 31% (table 1). In the jugular blood, the specific activity of methionine reached a plateau before 5 h after the first meal; in the portal blood 8 h after birth it represented 75% of the specific activity entering the small intestine. The blood methionine flux was calculated according to two methods: from whole-body protein synthesis rates and methionine catabolism and from the irreversible loss of blood methionine (table 2).(ABSTRACT TRUNCATED AT 250 WORDS)

[Effect of dietary protein levels on the amount of blood free amino acids in lambs]. / Influence du taux azoté la ration sur l'aminoacidémie libre de l'agneau.

Growing lambs were fed increasing amounts of nitrogenous matter (12 to 20 per 100 on dry matter basis). Free amino acid and urea levels were measured in blood of 37 kg lambs. When crude protein content of diets exceeded 16 per 100, the catabolism of several essential amino acids increased. Methionine, lysine and histidine seemed to be limiting for growing lambs.

[Influence of low energy alimentation and nitrogen on free amino acids in the blood of dairy cows at the beginning of lactation]. / Influence d'une sous-alimentation énergétique et azotée sur l'aminoacidémie libre de vaches laitières en début de lactation.

Dairy cows are fed low energy and adequate nitrogen (T), or low energy and low nitrogen (C) diets. Blood free aminoacids, urea and glucose are estimated. The relations between gluconeogenesis, protein catabolism, liveweight and milk production are discusssed.

[Comparison of the digestive utilization of methionine, of its hydroxylated analog, and of sodium sulfate in goats using 35s compounds]. / Etude comparée de l'utilisation digestive de la méthionine, de son analogue hydroxylé et du sulfate de sodium chez la Chèvre à l'aide de composés 35S

35S and 35S free and protein bound amino acids were estimated in goats' abomasal contents and blood after ruminal injections of sulfer labelled compounds: methionine, methionine hydroxy analog (M.H.A.) and sodium sulfate. 35S incorporation into microbial and plasma proteins was higher with methionine than with M.H.A. or sulfate. 35S.M.H.A. utilisation seems to be less different from Na2 35SO4 utilisation than from 35S methionine utilisation.