The role of glucocorticoids in regulating the rate of muscle protein breakdown was evaluated by measuring excretion of N(tau)-methylhistidine during administration of various doses of corticosterone to adrenalectomized rats. Groups of rats received daily subcutaneous injections of 0, 0.2, 0.5, 1.0, 5.0 or 10.0mg of corticosterone/day per 100g body wt. for 7 days, followed by 3 days without hormone treatment, after which they were killed. A group with intact adrenal glands served as an additional control. All animals were pair-fed with the untreated adrenalectomized group. No significant differences were noted in growth rate or N(tau)-methylhistidine excretion between the intact or adrenalectomized control groups, or those given 0.2, 0.5 and 1.0mg of corticosterone, whereas growth ceased and N(tau)-methylhistidine excretion rose markedly in the groups receiving 5 and 10mg of corticosterone. After these two high doses of corticosterone, but not after lower doses, there was a loss of weight of the gastrocnemius muscle per 100g of final body wt., but not of the soleus and extensor digitorum longus muscles. The two highest doses of corticosterone also resulted in an increase in liver weight per 100g of final body wt. Lower doses of corticosterone did not cause these changes. Plasma corticosterone concentrations, measured on the final day of injection and again at the time of killing, were decreased to near zero by adrenalectomy and were little raised by doses of 0.2 and 0.5mg daily, but were increased to within the normal range by the 1mg dose. At 5 and 10mg doses, plasma corticosterone concentrations were sustained at 2-3 times those of intact rats, and thus in the range reported for rats exposed to severe stress. Rats given 5 and 10mg doses of corticosterone had glycosuria, and showed considerably elevated concentrations of insulin in the plasma. It is concluded that plasma concentrations of glucocorticoids within the normal range do not regulate the rate of muscle protein breakdown, whereas excessive plasma concentrations of corticosteroids, equivalent to those observed in severe stress, can accelerate muscle protein breakdown.

To investigate the effect of insulin-like growth factor (IGF) peptide infusion on the gastrointestinal tract, female rats (115 g, 6/group) were treated for 14 days with IGF-I or long R (LR3IGF-I; 0, 44, 111, or 278 micrograms/day) delivered by osmotic minipumps. Both peptides induced a dose-dependent increase in gastrointestinal tissue weight. Total gut weight, small intestinal weight, and small intestinal length increased by 43, 47, and 13%, respectively, after treatment with 278 micrograms/day of LR3IGF-I. Crypt depth and villus height increased after peptide treatment with an associated increased crypt cell population (+33%), cells per villus column (+34%), and villus cell density (+20%). Proportional increments in proliferating cell nuclear antigen labeling and an unaltered crypt growth fraction indicated that the balance between the proliferative and maturation compartment of the crypt was maintained. Fecal nitrogen excretion was significantly reduced in rats treated with LR3IGF-I, suggesting an increased absorptive capacity of the duodenum. The enhanced potency of LR3IGF-I supports previous findings that the gut is especially responsive to analogues with reduced binding affinity to IGF-binding proteins.

Effects of insulin-like growth factor I (IGF-I) administration and that of the truncated analogue des-(1-3)IGF-I have been examined in 170-g rats over a 7-day period after surgery to remove 80% of the jejunum plus ileum. The doses administered via osmotic infusion pumps were 0.96 and 2.4 mg.kg-1.day-1 IGF-I and 0.96 mg.kg-1.day-1 des-(1-3)IGF-I. All groups lost weight on the day after surgery, but over the next 3 days the des-(1-3)IGF-I and high-dose IGF-I groups stabilized better and subsequently gained significantly (P less than 0.05) more weight than the vehicle or low-dose IGF-I groups over the last 3 days. The weight gains (mean +/- SE) for the groups over this last 3-day period were 14.0 +/- 1.7, 14.4 +/- 2.9, 21.9 +/- 1.7, and 20.8 +/- 1.0 g for the vehicle, low-dose IGF-I, high-dose IGF-I, and des-(1-3)IGF-I groups, respectively. The nitrogen balances over the last 3 days for the high-dose IGF-I and des-(1-3)IGF-I groups, at 242 +/- 14 and 217 +/- 13 mg/d, respectively, were significantly (P less than 0.05) more positive than the control group at 153 +/- 21 mg/d. These differences could at least partially be explained by changes in muscle protein breakdown, as assessed by 3-methyl-L-histidine excretion. The kidneys were heavier in all treatment groups and the thymus after administration of des-(1-3)IGF-I.(ABSTRACT TRUNCATED AT 250 WORDS)

The administration of insulin-like growth factor-I (IGF-I) via subcutaneously implanted osmotic pumps partially reversed a catabolic state produced by the co-administration of 20 micrograms of dexamethasone/day to 150 g male rats. Marked dose-dependent effects on body weight and nitrogen retention were produced, with the highest IGF-I dose, 695 micrograms/day, giving a 6 g increase in body weight over 7 days, compared with a 19 g loss in the dexamethasone-only group and an 18 g gain in pair-fed controls. Two IGF-I analogues that bind poorly to IGF-binding proteins, the truncated form, des(1-3)IGF-I, and a variant with an N-terminal extension as well as arginine at residue 3, LR3IGF-I, were approx. 2.5-fold more potent than IGF-I. The response with LR3IGF-I was particularly striking because this peptide binds 3-fold less well than IGF-I to the type 1 IGF receptor. The increased potencies of the IGF-I variants may relate to the substantially increased plasma levels of IGF-binding proteins, particularly IGFBP-3, produced by the combined treatment of dexamethasone with IGF-I or the variants. These binding proteins would be expected to decrease the transfer of IGF-I, but not that of the variants, from blood to tissue sites of action. Measurements of muscle protein synthesis at the end of the treatment period and muscle protein breakdown by 3-methylhistidine (3MH) excretion throughout the experiment indicated coordinate anabolic effects of the IGF peptides on both processes. Thus 3MH excretion was decreased at the highest IGF-I dose from 83.5 +/- 4.2 (S.E.M.) mumol/kg per 7 days to 65.1 +/- 2.2, compared with 54.9 +/- 1.2 in the pair-fed controls. Part of this response in 3MH excretion may have reflected a decrease in gut protein breakdown, because IGF-I and especially the IGF analogues increased the gut weight by up to 45%. Notwithstanding the effects on protein synthesis and breakdown, the fractional carcass weights remained low in the IGF-treated groups, although the increase in total carcass weight reflected nitrogen rather than fat gain. The dexamethasone-induced changes in liver, spleen and heart weight were restored towards normal by the IGF treatment. The experiment demonstrates the potential of IGF-I treatment of catabolic states and especially the value of modified forms of growth factors that bind weakly to IGF-binding proteins.

Nine 'large-eating ' (approximately 12 MJ/d) and nine 'small-eating ' (approximately 5.3 MJ/d) women were selected from the population on the basis of diet and activity diaries. At rest and in the postabsorptive state the rate of oxygen consumption ( poJ/kg fat-free mass (FFM) and rate of carbon dioxide production (&,)/kg FFM were 9-17% higher (P < 0.05) in the 'large-eaters' than in the 'smalleaters'. As energy expenditure was increased by walking at 2.4,3.9 and 5.4 km/h the differences between the two experimental groups for both & / k g FFM and ec0Jkg FFM were decreased to negligible values, but energy expended on a body-weight basis (MJ/kg per min) remained significantly higher (510 %) in 'large-eaters'. Oral temperature was also consistently higher (up to 0.5") in this group both at rest and during sitting, standing and walking activities. Although the average thermic effect of a standardized liquid meal tended to be higher (27 YO ; not significant) in the 'small-eaters', the other results demonstrate that the 'large-eating' females had a markedly higher rate of energy expenditure at rest and during light physical activities.Energy metabolism : Indirect calorimetry : Women

We have examined the influence of nutrition on plasma IGF-I, IGF-II and IGF-binding protein (IGFBP) levels and on hepatic IGF-I gene expression in young meat-type chickens. Plasma IGF concentrations were measured by using RIA with recombinant chicken IGFs as standards. In chickens fed the control diet containing 200 g/kg dietary protein ad libitum for 7 days, plasma IGF-I concentrations increased significantly from those found in the initial control group. Food restriction for either 4 or 7 days decreased plasma IGF-I by 30% from the initial control. When chickens were refed ad libitum for 3 days after 4 days of restricted feeding, plasma IGF-I levels recovered to those of the control birds fed ad libitum. In chickens eating a low protein diet (100 g/kg protein), the plasma IGF-I tended to be lowered but the decrease was not significant. Although the intensity of IGF-I and beta-actin mRNA bands protected in the RNase protection assay was changed by nutrition, no statistical effect of nutrition on the ratio of IGF-I to beta-actin was observed. The nutritional treatments had no effect on plasma IGF-II concentrations. Western ligand blot and chromatographic analyses were used to investigate the influence of nutrition on IGFBP profiles. Both IGF-I and IGF-II ligands in the Western ligand blot revealed the most intense binding at 30 kDa for plasma obtained from chickens with restricted food intake. The 30 kDa band also appeared at a lower intensity in the group fed a low protein diet but not in any other groups. These observations were confirmed by neutral gel chromatography. The chicken IGF-II ligand revealed an intensely labelled band corresponding to 75 kDa and this was not affected by nutrition. IGF-I and IGFBP concentrations in the plasma of young broiler chickens were influenced by nutritional state but IGF-II concentrations were not. The lack of a response in circulating IGF-II levels may have been due to the presence of high concentrations of a 75 kDa specific binding protein which did not respond to nutrition in this experiment.

1. Urinary excretion of 3-methylhistidine and creatinine have been measured in normal male and female humans ranging in age from pre-term neonates to 68 years to assess changes in the rate of muscle protein degradation and in muscle mass. 2. The 3-methylhistidine excretion by six adult men was measured before and after subjects were transferred to meat-free diets. It was established that a meat-free diet should be eaten for a minimum of 3 days before urine collection to eliminate exogenous sources of 3-methylhistidine. 3. The 3-methylhistidine/creatinine excretion ratio declined about twofold between normal full-term birth and maturity. The ratio in pre-term neonates was higher than for full-term neonates. 4. The variability of 3-methylhistidine/creatinine ratios between individual untime urine samples within a subject is similar to the variability between different subjects with total daily collections. 5. The 3-methylhistidine content of human muscle averaged 3.63 +/- 0.06 micromol/g for subjects aged between 4 and 65 years with protein accounting for 20.4 +/- 0.8% of muscle weight. These values are used to relate 3-methylhistidine excretion to muscle protein degradation. 6. It is concluded that control groups must be matched by age and sex to the group being examined. Where creatinine excretion is not perturbed it can be used as a reference base for comparisons of 3-methylhistidine excretion to indicate the average fractional degradation rates of muscle protein.

The ability of insulin-like growth factor-I (IGF-I) to protect against losses of body protein during periods of dietary nitrogen restriction has been evaluated in young rats. Recombinant human IGF-I was administered by osmotic pumps at dose rates of 0, 1.2 or 2.9 mg/kg per day over a 7-day period beginning with the transfer of animals from an 18% to a 4% protein diet. A fourth group received the potent truncated IGF-I analogue, des(1-3)IGF-I, at a dose of 1.2 mg/kg per day over a comparable 7-day period. Plasma IGF-I levels were reduced by 60% following nitrogen restriction, a reduction that was partly prevented by IGF-I administration, especially at the higher dose, but not measurably by des(1-3)IGF-I. The major IGF-binding protein circulating in blood, IGFBP-3, demonstrated a similar pattern of change. A significant (P less than 0.05) protection of body weight was achieved in the low dose IGF-I and des(1-3)IGF-I groups, but only after differences in food intake had been eliminated by analysis of covariance. Nitrogen balances were not significantly different unless analysis of covariance was used to adjust for the nitrogen intakes, whereupon all treatment groups showed improved balance, especially the animals treated with the low IGF-I dose and des(1-3)IGF-I (both P less than 0.01). The rate of muscle protein breakdown calculated from the urinary excretion of 3-methyl-histidine was not significantly altered by the treatments, but fell progressively throughout the 7 days. The fractional rate of muscle protein synthesis measured on the final day was increased by 31,26 and 21% respectively by the low and high doses of IGF-I and by des(1-3)IGF-I. Organ weights (g/kg body weight) showed no effects of IGF-I treatment except for 16% increases in the weight of kidneys in the high dose IGF-I and the des(1-3)IGF-I groups. Carcass analyses demonstrated higher water and lower fat contents (all P less than 0.01) in the same groups. These results suggest that exogenous IGF-I and especially des(1-3)IGF-I can partly protect body protein reserves during nitrogen restriction.