Urea kinetics in humans at two levels of exercise intensity

A primed constant infusion of [15N2]urea was used to quantify the response of urea production to exercise at 40 and 70% maximal oxygen consumption on a treadmill. Total urea production, urea production from recycled N, urea production from nonrecycled N, and urea N recycled back into body protein were calculated. Most components of urea kinetics were unaffected by exercise at either intensity. The rate of urea reincorporated into protein was significantly increased during exercise and recovery at both levels of exercise. We conclude that exercise does not stimulate urea production but that there may be an accelerated reincorporation of urea N back into body protein.     

Hypophosphatemia and glucose intolerance: evidence for tissue insensitivity to insulin

Although hypophosphatemia is commonly present in diabetics, little is known about its isolated effects on glucose and insulin metabolism. We therefore investigated glucose metabolism in six nondiabetic subjects with chronic hypophosphatemia. When glucose was infused to maintain a constant hyperglycemic level (125 mg per deciliter [6.9 mmol per liter] above basal levels), the glucose infusion rate was 36 per cent less in the hypophosphatemic group than in controls (4.90 +/- 0.34 mg per kilogram of body weight per minute vs. 7.64 +/- 0.37, P < 0.001), although responses to endogenous insulin were similar. When exogenous insulin was infused at a constant rate to maintain an insulin level about 100 microU per milliliter (718 pmol per liter) above basal levels and glucose was infused as necessary to maintain fasting glucose levels, the infusion rate of glucose was 43 per cent lower in the hypophosphatemic group than in controls (3.80 +/- 0.58 mg per kilogram per minute vs. 6.70 +/- 0.33, P < 0.001), although the clearance rate of insulin was similar in both groups. These results indicate that hypophosphatemia is associated with impaired glucose metabolism in both the hyperglycemic and euglycemic states, and that this associated primarily reflects decreased tissue sensitivity to insulin. (N Engl J Med. 1980; 303; 1259-63.).     

Purification by column chromatographies of beta-amyloid precursor proteins and their association with other 95 kDa protein in rat brain

For verifying catabolic states in insulin-dependent patients and dogs the method estimating urea production rates with 13C and with doubly 15N labeled urea, respectively, has been established. For a fast steady state of urea tracer dilution, a prime of 600 times the continuous infusion rate had to be injected. Urea was isolated from plasma samples by protein precipitation and cation exchange chromatography with a consecutive derivatization of the dried urea fraction (trimethylsilyl derivatives). The masses of the fragment ions m/z 189 (14N14N), 190 (14N15N) and 191 (15N15N) urea are monitored to estimate the [15N2] urea frequency in the overall body urea pool in mol percent excess (MPE). 1 to 15 ng of derivatized urea were measured efficiently. An excellent correlation between expected standard and measured MPE (r = 0.9977) was achieved from solutions containing 1 to 7% [15N2]urea. The interassay coefficient of variation amounted to < 10% for a [15N2]urea portion of > or = 3%. Normoglycemic diabetic patients who were treated with insulin overnight showed significantly higher urea production compared to healthy controls (9.22 +/- 2.07 vs. 5.4 +/- 0.32 mumol.kg-1.min-1; p < 0.05). Measurements in chronic diabetic dogs proved an increased rate of amino acid catabolism (+20% urea production) in systemic versus portal application of insulin in paired studies. This increased nitrogen load in diabetics may accelerate progression of diabetic nephropathy. Thus, the established stable isotope technique may serve as a sensitive and useful indicator of amino acid catabolism in clinical and experimental research.     

Treatment of heterotransplanted Hodgkin''s tumors in SCID mice by a combination of human NK or T cells and bispecific antibodies

OBJECTIVE: To investigate the kinetics of body nitrogen (N) excretion during 24 h glucose infusion (relating glycemia with insulin supply) and during subsequent 24 h saline infusion in injured patients during a full blown stress reaction. To define the lag time between the start of the withdrawal of glucose and insulin infusion, and the modification in the N loss from the body, and the time span to reach the maximum effect and its size. The knowledge of these variables is mandatory to plan short term studies in critically ill patients, while assuring the stability of the metabolic condition during the study period, and also to assess the possible weaning of the effect on protein breakdown during prolonged glucose and insulin infusion. DESIGN: 24-36 h after injury, patients were fasted ( < 100 g glucose) for 24 h (basal day). Thereafter, a 24 h glucose infusion in amount corresponding to measured fasting energy production rate (EPR), clamping glycemia at normal level with insulin supply followed by 24 h saline infusion, was performed. Total N, urea and 3-methyl-histidine (3-MH) in urine were measures on 4 h samples starting from 20th h of the basal day. SETTING: Multipurpose ICU in University Hospital. PATIENTS: 6 consecutive patients who underwent accidental and/or surgical injury, immediately admitted for respiratory assistance (FIO2 < 0.04). Excluded patients were those with abnormal nutritional status, cardiovascular compromise and organ failures. MAIN RESULTS: Patients showed a 33% increase in measured versus predicted fasting EPR and a consistent increase in N and 3-MH urinary loss. An infusion of glucose at 5.95 +/- 0.53 mg/kg x min (97.20 +/- 0.03% of the fasting measured EPR) with 1.22 +/- 0.18 mU/kg x min insulin infusion reduced N and 3-MH loss after a time lag of 12 h. The peak decrease in body N (-36%) and 3-MH loss (-38%) was reached during the first 12 h of glucose withdrawal period. Thereafter, during the following 12 h, the effect completely vanished confirming that it is therapy-dependent and that the metabolic environment of the patients did not change during the three days study period. CONCLUSION: 24 h glucose withdrawal reduces N and 3-MH loss injured patients, the drug-like effect is maintained during the first 12 h of withdrawal and thereafter disappears. The study suggests that at least a 24 h study period is necessary when planning studies exploring energy-protein metabolism relationship in injured patients, and, again 24 h before changing protocol in a crossover study.     

Non-insulin- and insulin-mediated glucose uptake in dairy cows

Four mid-lactation Holstein dairy cows (mean milk yield on day of experiments 26.1 kg/d) were used in a series of experiments to establish the contribution of non-insulin-mediated glucose uptake to total glucose uptake at basal insulin concentrations. A secondary objective was to determine whether somatostatin affects the action of infused insulin. In part I of the experiment a primed continuous infusion [6,6-2H]glucose (45.2 micrograms/kg per min) was begun at time 0 and continued for 5 h. After 3 h of [6,6-2H]glucose infusion (basal period) a primed continuous infusion of insulin (0.001 i.u./kg per min) was administered for 2 h. Coincidental with the insulin infusion, normal glucose was also infused in order to maintain the plasma glucose concentration at euglycaemia. Part II of the experiment was the same as part I except that somatostatin was infused for 2 h (0.333 micrograms/kg per min) instead of insulin. In part III of the experiment both insulin and somatostatin were infused for the final 2 h. Plasma insulin levels were increased by insulin infusion (to 0.1476 to 0.1290 i.u./l for parts I and III respectively) and were reduced by somatostatin infusion in part II (to 0.006 i.u./l) relative to the basal periods (mean 0.021 i.u./l). Glucose uptake during somatostatin infusion (2.50 mg/kg per min; part II) was 92.0% of that observed in the respective basal period (2.72 mg/kg per min). Circulating insulin levels were much lower than the dose of insulin that causes a half maximal effect on glucose uptake (0.06-0.10 i.u./l for ruminants); consequently insulin-mediated glucose uptake was probably absent in part II. Secondly, glucose uptake following insulin only infusion (4.05 mg/kg per min) was significantly lower than that observed when insulin plus somatostatin was infused (4.69 mg/kg per min), indicating that somatostatin either directly or indirectly enhanced the action of insulin on glucose uptake.     

Urea, glucose and alanine kinetics in man: effects of glucose infusion

1. The simultaneous effects of an intravenous glucose infusion on plasma urea, glucose and alanine kinetics were investigated in normal post-absorptive man. 2. The primed constant intravenous infusion of compounds labelled with stable isotopes, [15N2]urea, [6-2H]glucose and [3-13C]alanine, was used. 3. The rate of appearance of glucose and urea in the plasma was rapidly reduced by the 17.7 mumol min-1 kg-1 glucose infusion. 4. In contrast, during the glucose infusion there was an increased rate of appearance of alanine in the plasma, and an increased percentage of glucose carbon atoms derived from alanine. 5. Reduced production of glucose and urea during the glucose infusion was not due to decreased gluconeogenesis from alanine.     

Synchronization of ruminal degradation of supplemental carbohydrate with pasture nitrogen in lactating dairy cows

Twelve Holstein cows in early to midlactation (8 fitted with ruminal cannulas) were used to test the hypothesis that the synchronization of the rate of ruminal degradation of supplemental carbohydrate and N from fresh pasture would increase the amount of N retained for growth and milk production. A concentrate based on ground shelled corn was fed either at the time that pasture was fed at 0900 and 1700 h (synchronous) or 4 h after pasture was fed at 1300 and 2100 h (asynchronous). The crossover design included a 6-d adjustment period and a 10-d milk sampling period. Nitrogen balance was determined during the last 5 d of each period. Cows fed the synchronous diet had a lower mean ruminal pH. Peak ammonia concentration at 3 and 5 h after pasture feeding in the morning was reduced by approximately 33% of values obtained from cows fed the asynchronous diet. The diurnal pattern of blood urea N concentration was similar to that of ruminal ammonia, but mean daily concentrations of blood urea N did not differ between diets. Nitrogen retained for milk production and growth was not influenced by diet, and no differences in milk production, composition, or efficiency were observed. Based on changes in ruminal concentrations of ammonia, synchronous ruminal release of supplemental carbohydrate with pasture N appeared to improve the capture of ruminal N; however, these changes were transient and did not change the N status or performance of dairy cows.     

Effect of amino acid supplementation on whole-body protein turnover in Holstein steers

We used the [15N]glycine single-dose urea end-product technique to measure whole-body protein turnover in six Holstein steers (250 +/- 18 kg). Steers were implanted with Revalor-S and continuously infused abomasally with water (4 L/d) or amino acids (AA; in 4 L/d water) in a crossover experiment (two 14-d periods). The AA infusion contained the following (g/d): lysine (5.3), methionine (3.3), threonine (3.2), tryptophan (1.0), histidine (2.1), and arginine (5.5). Steers were fed a diet containing 85% rolled corn, 10% prairie hay, and 1.1% urea (DM basis) at 2.16% of body weight. Nitrogen retention tended (P = .15) to increase with AA infusion, from 27.9 to 32.9 g N/d. Amino acid infusion numerically increased whole-body protein turnover from 168.6 to 183.2 g N/d, protein synthesis from 152.6 to 169.3 g N/ d, and protein degradation from 124.7 to 136.4 g N/d. Enhanced protein accretion may have resulted from a larger increase in protein synthesis than in degradation. The tendency for increased N retention is interpreted to suggest that the implanted, lightweight Holstein steers fed a corn-urea diet in our study were able to respond to AA supplementation, suggesting that at least one of the infused AA was limiting in the basal diet. Protein turnover data suggest that cattle, like other animals, may increase protein synthesis and protein degradation in response to supplementation with limiting AA. The [15N]glycine single-dose urea end-product technique for measuring whole-body protein turnover in cattle may be useful.     

Effects on nutrient and hormonal profile of long-term infusions of glucose or insulin plus glucose in cows treated with recombinant bovine somatotropin before peak milk yield

Ten Holstein cows were treated with 30.9 mg.d-1 of recombinant bST from 15 to 41 d of lactation. The Latin square design included three infusion periods of 6 d each with 3 d of rest between infusion periods. Infusions were physiological saline, glucose (50 g.h-1), and insulin plus glucose (12.5 IU.h-1 + 50 g.h-1). Blood was collected continuously during the last 24 h of each infusion period. Statistical analyses of data for energy balance, milk yield, and DMI were performed on the last 3 d of each infusion period. Production data before and after infusions (i.e., no recombinant bST) estimated that recombinant bST increased milk yield of cows infused with glucose and saline by 3.1 and 3.6 kg.d-1, respectively. Net energy intake was not affected by infusion, but glucose infusion resulted in higher BW loss than did saline infusion (2.33 vs. 0.08 kg.d-1, respectively), and insulin plus glucose infusion resulted in BW gain (0.65 kg.d-1). Milk yield was 39.9, 39.6, and 37.6 kg.d-1 for cows infused with saline, glucose, and insulin plus glucose, respectively. The insulin plus glucose infusion increased milk protein 11 and 14% compared with response to saline and glucose infusions, respectively; no change occurred in the proportion of casein and whey proteins. Serum bST was increased 109% with exogenous recombinant bST. Serum IGF-I was lower for cows infused with glucose than for those infused with saline (21.03 vs. 27.44 ng.ml-1) and increased to 46.55 ng.ml-1 for cows infused with insulin plus glucose. Serum concentrations of insulin and glucose were 13.7 and 56.7, 18.5 and 61.9, and 30.5 muIU.ml-1 and 39.4 mg.dl-1 for cows infused with saline, glucose, and insulin plus glucose, respectively. The results of this study suggest that low concentrations of plasma insulin in early lactation may limit the IGF-I response to recombinant bST (uncoupling). Despite higher IGF-I, milk yield was lower, probably as a result of low blood glucose. These results suggest that, in early lactation, insulin is still anabolic because the BW gain of cows increased. However, milk yield was still higher than that for cows in late lactation with similar insulin concentrations.     

Oscillatory insulin secretion after pancreas transplant

In vivo studies of beta-cell secretory function have demonstrated the existence of rapid insulin oscillations of small amplitude recurring every 8-15 min in normal subjects. This study evaluated the effects of pancreas transplant on rapid insulin oscillations. Samples for glucose, insulin, and C-peptide were drawn during constant glucose infusion at 2-min intervals for 90 min from six successful Px patients with type I diabetes mellitus, from six normal nondiabetic control subjects, and from three Kx subjects. A computerized algorithm (ULTRA) was used for pulse detection. In the Px group, the average insulin pulse period was significantly shorter than in both the control and Kx groups (Px 8.1 +/- 0.5, control 12.5 +/- 0.7, Kx 12.4 +/- 0.5 min, P < 0.0005). By contrast, the C-peptide pulse periods (Px 16.8 +/- 2.3, control 14.7 +/- 1.2, Kx 15.3 +/- 1.5 min) were similar in the three groups. Spectral analysis confirmed that the frequency of the insulin pulses was increased in the Px group. The absolute amplitude of the insulin pulses was greater in the Px group (P < 0.001) while the amplitude of the C-peptide pulses did not differ between the groups. Cross-correlation analysis demonstrated maximal correlation coefficients at a lag of 0 min between insulin and C-peptide (control r = 0.33, P < 0.0001; Kx r = 0.17, P = 0.06) and between insulin and glucose (control r = 0.21, P < 0.001; Kx r = 0.20, P < 0.02) in the control and Kx groups, respectively, whereas no significant correlations were observed at any lag in the Px group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of hyperinsulinemia under the euglycemic condition on calcium and phosphate metabolism in non-obese normotensive subjects

The effect of acute insulin infusion on the metabolism of calcium (Ca) and phosphate (P) was examined in 17 healthy subjects. They were hospitalized and kept on a constant diet for 5 days, and an euglycemic hyperinsulinemic glucose clamp was applied. Synthetic human insulin was infused at the rate of 40 mU/m2/min for 2 hr, and glucose was also infused to maintain basal glucose levels of each subject. The control study was performed in 8 of the 17 subjects, into whom 10% xylitol was infused for 2 hr at the rate of 100 ml/hr. The plasma insulin concentrations were 7.94 +/- 0.35 and 62.3 +/- 14.3 mU/liter before and after the glucose clamp technique, but serum free Ca ion was increased significantly (p < 0.05), and serum P and serum parathyroid hormone (PTH) were decreased significantly (p < 0.001). Creatinine clearance did not change during the glucose clamp technique. Urinary excretion of Ca (UCaV) was significantly higher after the glucose clamp than the control study. Fractional excretion of Ca (FECa) was increased significantly (p < 0.05), and urinary excretion of P (UPV) and fractional excretion of P (FEP) were decreased significantly (p < 0.05) under the hyperinsulinemic condition. The results suggested that, under the conditions of euglycemic hyperinsulinemia by glucose clamp technique, insulin increased the serum free Ca ion, and as a result, PTH was suppressed. Decreased PTH might induce calciuresis and enhance tubular P reabsorption under hyperinsulinemia. Insulin increased serum free Ca ion might relate to the vasodilating action of insulin by its decrease of intracellular free Ca ion in vascular smooth muscle.     

Effects of chronic intrahypothalamic infusion of insulin on food intake and diurnal meal patterning in the rat.

In Experiment 1, rats were chronically infused with insulin (2.7, 27, or 270 ng/hr) or 0.9% saline into the ventromedial (VMH), medial perifornical (PF), or lateral (LH) hypothalamus. VMH infusions of insulin caused a significant, dose-dependent decrease in food intake and body weight; PF infusion of insulin was less effective, but significant; whereas LH infusions of insulin were ineffective. In Experiment 2, rats were chronically infused with insulin (0.54 ng/hr) or 0.9% saline into the VMH, paraventricular (PVN), or posterior (PN) hypothalamic nucleus. Subjects that received VMH or PN infusions of insulin failed to regain weight lost as a result of surgery even 2 weeks after infusion; subjects that received PVN infusions of insulin regained their preoperative weights faster than did controls. All of the groups that received insulin significantly increased their daytime food intake during the infusion period and decreased their night food intake slightly; 24-hr food intake remained unchanged. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Determination of amino- and amide-15N glutamine enrichment with tertiary butyldimethylsilyl derivatives

We have developed a simple and rapid method for the selective synthesis of tetra-tertiarybutyldimethylsilyl (TBDMS) glutamine, which allows the simultaneous quantitation of glutamine (2-15N) and (5-15N) isotopic enrichment by selected ion monitoring (SIM) gas chromatographic/mass spectrometric analysis. The tetra-TBDMS glutamine (4S-gln) has an electron impact mass fragment at m/z 258 that contains only the amino-N and larger fragments (e.g., at m/z 545) that have both nitrogens. Derivatization with acetonitrile (ACN) and N-methyl-N-(tert-butyldimethylsilyl)trifluoroacetamide yields primarily tri-TBDMS glutamine (3S-gln) and quantities of 4S-gln too small to allow accurate SIM and tracer/tracee ratio determinations. However, when N,N-dimethylformamide, a more polar aprotic solvent, was substituted for ACN and the sample was heated for 30 min at 125 degrees C, greater than 80% of derivatized glutamine appeared as 4S-gln. Derivatized plasma samples that had been mixed with amide- and/or amino-15N glutamine and analyzed by SIM demonstrated strong agreement (r > or = 0.998, p = 0.0001) between theoretical and observed enrichment values for the 4S-gln fragments at m/z 258 and 545. Deamidation of glutamine to glutamate is negligible during sample processing and analysis. This procedure will facilitate the investigation of the specific sources and fates of glutamine amide and amino nitrogen as well as stable isotope studies involving amino acid transamination, ammonia clearance, urea production and other areas of nitrogen metabolism.     

Effect of hepatic vagotomy on plasma insulin levels during fasting in rats

The present investigation was designed to evaluate the effect of a selective hepatic vagotomy (HV) on the insulin response in rats fasted for 24 h when blood glucose levels were or were not maintained by a constant glucose infusion. Rats were divided into three dietary groups: one group of normally fed rats, one group of 24-h fasted rats, and one group of 24-h fasted rats infused with glucose throughout the fasting period. Each of these groups was subdivided into HV and sham-operated (SHM) rats. Fasting without glucose infusion resulted in a significant (p < 0.05) decrease in plasma glucose, liver glycogen, and insulin concentrations and in a significant (p < 0.01) increase in beta-hydroxybutyrate and FFA concentration. Despite the maintenance of plasma glucose concentrations in the glucose-infused groups, the concentrations of liver glycogen and insulin were still decreased (p < 0.01) and the concentrations of beta-hydroxybutyrate were still increased (p<0.05) at the end of the fasting period. However, no significant differences in insulin or in beta-hydroxybutyrate concentration were found between HV and SHM rats. It is concluded that the decline in plasma glucose concentration during fasting does not totally explain the insulinopenic response to fasting, and that the liver, through the mediation of the hepatic vagus nerve, does not seem to contribute to insulinopenia in 24-h fasted rats.     

15N-NMR spectroscopy studies of ammonia transport and glutamine synthesis in the hyperammonemic rat brain

Ammonia transport and glutamine synthesis were studied in the hyperammonaemic rat brain in vivo using 15N-NMR spectroscopy at a plasma ammonia level of approximately 0.39 mM raised via an intravenous [15N]-ammonium acetate infusion. The initial slope of the time course of the summed cerebral 15N-labelled metabolites was used to determine the rate of ammonia net transport during hyperammonemia as 0.13 +/- 0.02 micromol/min/g (mean +/- SD; n = 5). Based on the total accumulation of glutamine and the 1:2 stoichiometric relationship between fluxes of four-carbon skeletons and nitrogen atoms, the rate of de novo glutamine synthesis through anaplerosis and subsequent glutamate dehydrogenase action was calculated to be 0.065 +/- 0.01 micromol/min/g. The rate of total glutamine synthesis was estimated to be 0.20 +/- 0.06 micromol/min/g (n = 5) by fitting the [5-15N]glutamine time course to a previously described model of glutamate-glutamine cycling between astrocytes and neurones. A large dilution was also observed in [2-15N]glutamine, which supports the glutamate-glutamine cycle as being an important pathway for neuronal glutamate repletion in vivo.     

Comparative pharmacokinetics and glucodynamics of two human insulin mixtures. 70/30 and 50/50 insulin mixtures

OBJECTIVE: To compare and contrast the pharmacokinetics and glucodynamics of two insulin mixtures, one of 50% NPH human insulin and 50% Regular human insulin (50/50) and one of 70% NPH human insulin and 30% Regular human insulin (70/30), in healthy male volunteers after subcutaneous administrations of 0.3 U/kg. RESEARCH DESIGN AND METHODS: We administered single doses of 50/50 and 70/30 insulins to 18 volunteers in a randomized crossover fashion. All subjects received 0.3 U/kg of each mixture separated by at least 7 days. Each dose was given after an overnight fast and during a glucose clamp to maintain a euglycemic state. We measured serum insulin and C-peptide concentrations through frequent blood sampling after each treatment. Pharmacokinetic measurements were calculated from insulin data corrected for C-peptide, including maximum insulin concentration (Cmax), time to maximum insulin concentration (tmax), terminal rate constant (beta), area under the curve from 0 to infinity (AUCinfinity0), and mean residence time (MRT). Pharmacodynamic measurements were summarized from C-peptide concentrations (minimum C-peptide concentration [Cmin], time to minimum C-peptide concentration [tmin], area between the C-peptide baseline and the C-peptide suppression curve [AOCc], absolute maximal difference from baseline [Sdiff] and glucose clamp measurements. The glucose clamp measurements included maximum infusion rates (Rmax) and time to Rmax (TRmax) from glucose infusion rate (GIR) documentation, as well as cumulative glucose infused during the first 4 h ((0)4Gtot) and total glucose infused (Gtot) during the study. RESULTS: For the pharmacokinetic assessment, statistically greater values of insulin Cmax and beta were found for the 50/50 mixture, whereas the 70/30 mixture had a greater MRT. Statistical differences were also detected in glucodynamics, with greater values of Rmax and (0)4Gtot found with the 50/50 mixture. Notably, differences were not detected for insulin AUCinfinity0 and Gtot values. CONCLUSIONS: Higher insulin concentrations and a greater initial response were present with the 50/50 mixture, but the two mixtures had equivalent bioavailability and cumulative effects. These results support use of the 50/50 mixture in situations where greater initial glucose control is required.     

RAGE mRNA expression in the diabetic mouse kidney

Data are accumulating that insulin acting in the central nervous system is a physiological regulator of food intake and body weight, presumably via its effect in the hypothalamus. The present study investigated whether infusion of a small dose of insulin into two major hypothalamic insulin-binding areas also has an effect on diet selection and behavior. At the beginning of the dark period, rats received local bilateral infusions of 4 microU of insulin or vehicle during 34 min into the arcuate (ARC) or paraventricular (PVN) nucleus of the hypothalamus. Consumption of carbohydrate (C)-, protein (P)-, and fat (F)-enriched food and time spent on certain behaviors (drinking, resting, grooming, rearing, exploring/sniffing) were assessed during the first nocturnal hour. In addition, 21-h diet selection was assessed. The percentage contribution of macronutrients (C/P/F) to total energy content of the C-, P-, and F-enriched diets was 71.9/17.2/10.9, 45.8/43.4/10.8, and 47.1/17.5/35.4, respectively. During the first hour, infusion of insulin into the PVN increased grooming behavior compared to infusion of the vehicle. Although infusion of insulin had no effect on diet selection during the first hour, insulin infused in the ARC caused a reduction in F-enriched food consumption and total intake of F (as a macronutrient) over the 21-h period without altering total food intake. Infusion of a higher dose of insulin (10 microU) into the third ventricle had no effect on any of the assessed parameters. The data are explained to indicate that insulin (being an indicator of a positive energy balance) adjusts body weight homeostasis by modulating the preference for fat, at least at the level of the ARC, but not at the PVN.     

Neuromuscular inhibitory effects of propofol evaluated by cat's evoked EMG

Neuromuscular inhibitory effects of propofol were investigated in 7 cats using muscular compound action potential (mCAP) elicited from the gastrocnemius muscle by sciatic nerve stimulation. A rectangular pulse of supramaximal strength and duration of 0.2 msec was applied serially to the sciatic nerve at 0.1 Hz during the experiments, and mCAPs obtained from the gastrocnemius muscle were recorded. After obtaining a constant mCAP amplitude, a series of 8 consecutive mCAPs (M 1-8) evoked by repetitive stimulation at 100 Hz was recorded, and M 8/M 1 amplitude ratio (M 8 amplitude/M 1 amplitude.100%) was calculated as the control value. After control variables had been obtained, initial dose of 2.5 mg.kg-1 of propofol was administered intravenously followed by continuous infusion with incremental dosages of 10, 20, 50 and 100 mg.kg-1.hr-1 of each 30 minute duration. The effects on mCAPs evoked by single and 8 consecutive repetitive stimulation were observed in each period. The depression of single mCAP amplitudes and fading responses in the M 8/M 1 amplitude ratios were not observed at any infusion rate. Our results suggest that propofol has no influence on neuromuscular transmission in cats when administered intravenously at 10 times of human clinical infusion doses.     

Whole body nitric oxide synthesis in healthy men determined from [15N] arginine-to-[15N]citrulline labeling

The rates of whole body nitric oxide (NO) synthesis, plasma arginine flux, and de novo arginine synthesis and their relationships to urea production, were examined in a total of seven healthy adults receiving an L-amino acid diet for 6 days. NO synthesis was estimated by the rate of conversion of the [15N] guanidino nitrogen of arginine to plasma [15N] ureido citrulline and compared with that based on urinary nitrite (NO2-)/nitrate (NO3-) excretion. Six subjects received on dietary day 7, a 24-hr (12-hr fed/12-hr fasted) primed, constant, intravenous infusion of L-[guanidino-15N2]arginine and [13C]urea. A similar investigation was repeated with three of these subjects, plus an additional subject, in which they received L-[ureido-13C]citrulline, to determine plasma citrulline fluxes. The estimated rates (mean +/- SD) of NO synthesis over a period of 24 hr averaged 0.96 +/- 0.1 mumol .kg-1.hr-1 and 0.95 +/- 0.1 mumol.kg-1.hr-1, for the [15N]citrulline and the nitrite/nitrate methods, respectively. About 15% of the plasma arginine turnover was associated with urea formation and 1.2% with NO formation. De novo arginine synthesis averaged 9.2 +/- 1.4 mumol. kg-1.hr-1, indicating that approximately 11% of the plasma arginine flux originates via conversion of plasma citrulline to arginine. Thus, the fraction of the plasma arginine flux associated with NO and also urea synthesis in healthy humans is small, although the plasma arginine compartment serves as a significant precursor pool (54%) for whole body NO formation. This tracer model should be useful for exploring these metabolic relationships in vivo, under specific pathophysiologic states where the L-arginine-NO pathway might be altered.     

Continuous estimation of CO2 production during exercise

A new method was developed for continuous isotopic estimation of human whole body CO2 rate of appearance (Ra) during non-steady state exercise. The technique consisted of a breath-by-breath measurement of 13CO2 enrichment (E) and a real-time fuzzy logic feedback system which controlled NaH13CO3 infusion rate to achieve an isotopic steady state. Ra was estimated from the isotope infusion rate and body 13CO2 enrichment which was equal to E at the isotopic steady state. During a non-steady state incremental cycle exercise (5 w/min or 10 w/min), NaH13CO3 infusion rate was successfully increased by the action of feedback controller so as to keep E constant.