Stable isotopic tracers as metabolic probes in exercise.
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Amino Acids↗
Biomedical subjects
R R Wolfe
Publications and source records attributed to R R Wolfe.
In vivo assessment of the metabolic alterations in glucagonoma syndrome.
Stable-isotope methodology and indirect calorimetry were used to evaluate metabolic abnormalities in a patient with glucagonoma syndrome manifested by 17% body weight loss, hypoaminoacidemia, and hyperglycemia. Energy expenditure (26 kcal/kg) was the same as that predicted by the Harris-Benedict equation. The rate of appearance (Ra) of intracellular leucine (2.70 mumol/kg/min), an index of protein breakdown, was normal, although the percentage of leucine flux oxidized (31%), an index of amino acid catabolism, was 50% greater than the normal mean value. Glucose Ra in plasma (12.9 mumol/kg/min), representing hepatic glucose production, and glycerol Ra in plasma (3.04 mumol/kg/min), a measurement of whole-body lipolysis, were 15% and 25% greater, respectively, than mean values found in normal volunteers. These results suggest that long-term alterations in energy, leucine, glucose, and lipid metabolism in patients with glucagonoma are minimal. However, small long-term metabolic alterations caused by glucagon excess, in conjunction with chronic negative energy balance, could be responsible for the weight loss, hypoaminoacidemia, and hyperglycemia observed in this patient population.
Adult↗
Generalized lipodystrophy: in vivo evidence for hypermetabolism and insulin-resistant lipid, glucose, and amino acid kinetics.
Stable isotope tracers and indirect calorimetry were used to evaluate whole-body energy, glucose, lipid, and amino acid metabolism in a patient with generalized lipodystrophy during basal conditions and in response to insulin therapy. The results were compared with those obtained in previous studies in normal volunteers. The basal rate of glucose production (33.7 mumol/kg.min) was three times higher than normal. The basal rate of glycerol appearance in blood, an index of lipolysis, was 60% greater than normal when expressed per kilogram body weight (3.82 mumol/kg.min), but was more than 10 times normal when expressed per kilogram body fat mass (123.2 mumol/kg.min) because of the marked decrease in body fat in our patient (3% of total body weight). Leucine rate of appearance, an index of protein breakdown, and nonoxidative leucine disposal, an index of protein synthesis, were also greater than normal. Resting energy expenditure (REE) was 30% greater than normal. The effect of insulin infusion on these metabolic parameters was markedly blunted. These metabolic abnormalities help explain many of the clinical findings such as hyperglycemia, hypertriglyceridemia, fat depletion, hepatomegaly, and steatosis observed in patients with lipodystrophy. Ineffective insulin function in many tissues appears to be an important factor in the pathophysiology of lipodystrophy.
Adult↗
Maximal voluntary exercise variables in children with postoperative coarctation of the aorta.
Thirty-one children with postoperative coarctation of the aorta underwent maximal graded bicycle ergometry using an electronically braked ergometer and the James protocol; 18 also underwent expiratory gas measurement using a mass spectrometer. Twenty-two age- and gender-matched normal subjects were used as a control group. The mean age at operation was 41 months and the mean age at evaluation was 134 months (mean follow-up interval 93 months). The original surgical repair was subclavian flap repair in 8 patients, end to end anastomosis in 21, patch aortoplasty in 1 patient and tubular graft in 1. Patients exercised until exhaustion and maximal exercise variables were obtained. The maximal voluntary peak heart rate was 183 beats/min (94.6% of predicted value), indicating excellent effort. Mean power was 111% of predicted value and, when measured, maximal oxygen consumption was 89% of predicted value with an anaerobic threshold at 63 +/- 3.5% of exercise time. The observed work variables were not different from values in the control group and were not affected by the type of repair. The mean peak systolic blood pressure was 152 +/- 7.6 mm Hg versus 147 +/- 5.7 mm Hg in the control group (p = NS). Patients who had associated intracardiac lesions had significantly lower maximal oxygen consumption (85 +/- 3% vs. 98 +/- 4% of predicted value). The results suggest that adequate cardiopulmonary function, normal or above average work capacity and normal exercise systolic blood pressure can be obtained in children with satisfactory repair of coarctation of the aorta performed before school age.
Adolescent↗
Longitudinal changes in basal hepatic glucose production and suppression during insulin infusion in normal pregnant women.
OBJECTIVE: The purpose of this study was to evaluate basal endogenous glucose production and suppression during insulin infusion in normal pregnant women. STUDY DESIGN: This prospective, longitudinal study was conducted at the Medical Center Hospital of Vermont. Six healthy women were evaluated before conception and at 12 to 14 and 34 to 36 weeks' gestation. Body composition was estimated by hydrodensitometry. Basal endogenous glucose production was estimated with a primed constant infusion of 6-6 2H2 glucose, and suppression of endogenous glucose production was estimated with insulin infusion during a hyperinsulinemic-euglycemic clamp. RESULTS: There was a significant (p = 0.02) 65% increase in fasting insulin concentration by late gestation. Moreover, there was a significant 30% (p = 0.0005) increase in basal endogenous glucose production (mg/min) with advancing gestation, which remained significant (p = 0.05) when corrected for fat-free mass. During insulin infusion endogenous glucose production was almost completely suppressed (i.e., > 90%) throughout gestation. CONCLUSIONS: There is a significant increase in basal endogenous glucose production at 34 to 36 weeks' gestation in spite of a significant increase in fasting insulin concentration. However, endogenous glucose production remains sensitive to insulin infusion throughout gestation.
Adult↗
Maximal voluntary work and cardiorespiratory fitness in patients who have had Kawasaki syndrome.
To assess the natural history of Kawasaki syndrome and its effect on maximal voluntary work and cardiorespiratory fitness, we performed cycle ergometry testing in 47 patients who had had the syndrome. Forty-one patients performed maximal effort as judged by achievement of 95% predicted heart rate response. Oxygen consumption, carbon dioxide production, and minute ventilation were performed in 23 patients. There was no difference in maximal voluntary work (total work, mean power) or maximal oxygen consumption between case subjects and control subjects. There were no differences between patients with and those without aneurysms. Serial exercise studies were performed in 10 patients; of these, two with initially normal exercise study findings had decreased maximal voluntary work and oxygen consumption with ischemic changes, and both were at high risk for the development of stenotic or occlusive coronary arteries. The other eight patients had normal cardiorespiratory reserve and no ischemic changes with serial studies. These results suggest that patients have normal cardiorespiratory fitness after Kawasaki syndrome. With the development of ischemic heart disease, they may have decreased cardiorespiratory reserve. Serial evaluation of cardiorespiratory fitness may demonstrate ischemic heart disease.
Adolescent↗
High-protein intake alters the response of fasting in normal human subjects.
Two groups of normal volunteers were studied for 5 d of dietary control followed by 3 d of fasting. One group (n = 5) was given a control diet of 0.9 g protein.kg-1.d-1 and the other group (n = 7) was given a high-protein (HP) diet (2.5 g protein.kg-1.d-1). Both groups received 175.56 kJ.kg-1.d-1 (42 kcal.kg-1.d-1). The HP diet but not the control diet caused a significant retention of nitrogen. Postabsorptive leucine kinetics as assessed with [1,2-13C]leucine were similar in the two groups. In the control subjects, the rate of nitrogen excretion did not change in response to fasting, but leucine oxidation increased. In contrast, nitrogen excretion progressively decreased with fasting after the HP diet. Leucine rate of appearance was increased after fasting after the HP diet but oxidation was not increased, meaning that the calculated rate of whole-body protein synthesis was higher than in the control group. The response to a short period of food deprivation is dependent on prior protein intake.
Dietary Proteins↗
Comparison of constant infusion and flooding dose techniques to measure muscle protein synthesis rate in dogs.
The exact relationship between the constant infusion and flooding dose techniques for measuring fractional muscle protein synthesis (FMPS) rates was determined in anesthetized dogs during the postabsorptive period, using different isotopes of leucine as tracers. First, [1,2-13C]- and [U-14C]leucine were given simultaneously by prime-constant infusion for 5 h to establish that both isotopes gave comparable FMPS rate values. The tracers gave almost identical FMPS rates and the values obtained after 3 and 5 h were not different. Next, the constant infusion and flooding dose methods were compared in the same animal by first measuring FMPS rate with a prime-constant infusion of [U-14C]leucine, then with a flooding dose of 376 mumol/kg [1,2-13C]leucine. The flooding dose method gave FMPS rates that were markedly greater than the constant infusion values. Finally, the effect of a 376 mumol/kg leucine flooding dose on FMPS rate was assessed using the prime-constant infusion of [U-14C]lysine. The large dose of leucine decreased FMPS rate by approximately 50% because acid-soluble muscle lysine (precursor) specific activity increased while the rate of incorporation of labeled lysine into protein decreased. These results indicate that the flooding dose method gives FMPS rates that are markedly greater than those obtained with the constant infusion method and that the latter method is not valid when a large dose of leucine is given.
Animals↗
Carbohydrate restriction regulates the adaptive response to fasting.
The importance of either carbohydrate or energy restriction in initiating the metabolic response to fasting was studied in five normal volunteers. The subjects participated in two study protocols in a randomized crossover fashion. In one study the subjects fasted for 84 h (control study), and in the other a lipid emulsion was infused daily to meet resting energy requirements during the 84-h oral fast (lipid study). Glycerol and palmitic acid rates of appearance in plasma were determined by infusing [2H5]glycerol and [1-13C]palmitic acid, respectively, after 12 and 84 h of oral fasting. Changes in plasma glucose, free fatty acids, ketone bodies, insulin, and epinephrine concentrations during fasting were the same in both the control and lipid studies. Glycerol and palmitic acid rates of appearance increased by 1.63 +/- 0.42 and 1.41 +/- 0.46 mumol.kg-1.min-1, respectively, during fasting in the control study and by 1.35 +/- 0.41 and 1.43 +/- 0.44 mumol.kg-1.min-1, respectively, in the lipid study. These results demonstrate that restriction of dietary carbohydrate, not the general absence of energy intake itself, is responsible for initiating the metabolic response to short-term fasting.
Adaptation, Physiological↗
Comparison of indirect calorimetry and a new breath 13C/12C ratio method during strenuous exercise.
A new stable isotope method for the determination of substrate oxidation rates in vivo is described and compared with indirect calorimetry at rest and during high-intensity exercise (30 min at 80-85% maximal O2 uptake capacity) in six well-trained cyclists. This method uses the absolute ratios of 13C/12C in expired air, endogenous glucose, fat, and protein in addition to O2 consumption and is independent of CO2 production (VCO2). Carbohydrate and fat oxidation rates at rest, calculated by both methods, were not significantly different. During exercise the breath 13C/12C ratio increased and reached a steady state after 15-20 min. Carbohydrate oxidation rates during exercise were 39.4 +/- 5.2 and 41.7 +/- 5.7 mg.kg-1.min-1 [not significant (NS)], and fat oxidation rates were 7.3 +/- 1.3 and 6.9 +/- 1.2 mg.kg-1.min-1 (NS), using indirect calorimetry, and the breath ratio method, respectively. We conclude that the breath 13C/12C ratio method can be used to calculate substrate oxidation under different conditions, such as the basal state and exercise. In addition, the results obtained by this new method support the validity of the underlying assumption that indirect calorimetry regards VCO2 as a reflection of tissue CO2 production, during exercise in trained subjects, even up to 80-85% maximal O2 uptake.
Adult↗
Stable isotope tracer analysis by GC-MS, including quantification of isotopomer effects.
In metabolic tracer studies it is frequently useful to infuse tracers that are differently labeled variants of the same molecule. These tracers are known as isotopomers. Analysis of the enrichment of each isotopic analogue can be accomplished by gas chromatography-mass spectrometry (GC-MS). However, the raw GC-MS data must be corrected to give the information required. This paper addresses how to transform the raw GC-MS data, consisting of relative abundance ratios at specific ion masses, into relative molar ratios of tracer and tracee molecules. Several correction factors are necessary. First, the background must be measured and corrected for, since it is always present in the sample. Second, the abundances in the spectrum of the labeled molecule are different from those in the unlabeled molecule, and this proportionality "skew" is corrected. A third correction factor accounts for the overlapping spectra of two or more isotopomers that cannot be measured independently. The final correction removes the "double vision" effect that may appear in some spectra due to the presence of (M - H)+ species.
Drug Stability↗
Muscle accounts for glucose disposal but not blood lactate appearance during exercise after acclimatization to 4,300 m.
We hypothesized that the increased blood glucose disappearance (Rd) observed during exercise and after acclimatization to high altitude (4,300 m) could be attributed to net glucose uptake (G) by the legs and that the increased arterial lactate concentration and rate of appearance (Ra) on arrival at altitude and subsequent decrease with acclimatization were caused by changes in net muscle lactate release (L). To evaluate these hypotheses, seven healthy males [23 +/- 2 (SE) yr, 72.2 +/- 1.6 kg], on a controlled diet were studied in the postabsorptive condition at sea level, on acute exposure to 4,300 m, and after 3 wk of acclimatization to 4,300 m. Subjects received a primed-continuous infusion of [6,6-D2]glucose (Brooks et al., J. Appl. Physiol. 70: 919-927, 1991) and [3-13C]lactate (Brooks et al., J. Appl. Physiol. 71:333-341, 1991) and rested for a minimum of 90 min, followed immediately by 45 min of exercise at 101 +/- 3 W, which elicited 51.1 +/- 1% of the sea level peak O2 uptake (65 +/- 2% of both acute altitude and acclimatization peak O2 uptake). Glucose and lactate arteriovenous differences across the legs and arms and leg blood flow were measured. Leg G increased during exercise compared with rest, at altitude compared with sea level, and after acclimatization. Leg G accounted for 27-36% of Rd at rest and essentially all glucose Rd during exercise. A shunting of the blood glucose flux to active muscle during exercise at altitude is indicated. With acute altitude exposure, at 5 min of exercise L was elevated compared with sea level or after acclimatization, but from 15 to 45 min of exercise the pattern and magnitude of L from the legs varied and followed neither the pattern nor the magnitude of responses in arterial lactate concentration or Ra. Leg L accounted for 6-65% of lactate Ra at rest and 17-63% during exercise, but the percent Ra from L was not affected by altitude. Tracer-measured lactate extraction by legs accounted for 10-25% of lactate Rd at rest and 31-83% during exercise. Arms released lactate under all conditions except during exercise with acute exposure to high altitude, when the arms consumed lactate. Both active and inactive muscle beds demonstrated simultaneous lactate extraction and release. We conclude that active skeletal muscle is the predominant site of glucose disposal during exercise and at high altitude but not the sole source of blood lactate during exercise at sea level or high altitude.
Acclimatization↗
Effect of recombinant human growth hormone on catabolic hormones and free fatty acids following thermal injury.
Severe burn injury elicits the release of catabolic hormones that contribute to negative nitrogen balance, protein wasting, and impaired wound healing. Previous studies have shown that burn patients receiving recombinant human growth hormone (rhGH) therapy have an increase in the rate of skin donor site healing and a shorter hospital stay. The mechanism by which rhGH exerts its effects, however, is not clearly understood. This study examines the effects of rhGH on circulating levels of catabolic hormones and nonesterified fatty acids in pediatric burn patients. Patients with greater than 40% total body surface area burn were randomly assigned to receive placebo (n = 8) or 0.2 mg/kg/day rhGH (n = 6) throughout their hospitalization. All patients had early morning blood samples assessed for catecholamines (CAT), cortisol, insulin, glucagon, and free fatty acid (FFA) levels during a period of hypermetabolism. No differences could be demonstrated in age, burn size, postburn day of evaluation, resting energy expenditure per kilogram, respiratory rate, heart rate, respiratory quotient, serum cortisol, and serum glucose between placebo- and rhGH-treated patients. The rhGH-treated group did show a significant elevation (p less than 0.05) in insulin-like growth factor-1 (55.9 +/- 14.5 vs. 168 +/- 23.7 mU/mL), total catecholamines (1,817 +/- 177 vs. 1,117 +/- 137 pg/mL), norepinephrine (1,257 +/- 121 vs. 867 +/- 113 pg/mL), epinephrine (385 +/- 175 vs. 147 +/- 36 pg/mL), insulin (32.8 +/- 3.3 vs. 25.0 +/- 3.0 mU/mL), glucagon (215 +/- 18 vs. 158 +/- 22 pg/mL), and free fatty acids (0.74 +/- 0.01 vs. 0.59 +/- 0.04 mEq/L) compared with the placebo group (data expressed as mean +/- SE).(ABSTRACT TRUNCATED AT 250 WORDS)
Burns↗
Hemodynamic effects of ketamine, hypoxia and hyperoxia in children with surgically treated congenital heart disease residing greater than or equal to 1,200 meters above sea level.
Little data are available on the hemodynamic effects of premedications and anesthetic agents on infants and children. Ketamine is the most frequently used anesthetic agent for cardiac catheterization procedures in pediatric patients with congenital heart disease. Previous reports both suggest and deny ketamine's pulmonary vasoreactive effects. Since the advent of sophisticated noninvasive equipment, one of the few indications for cardiac catheterization is to obtain accurate pressure data. If ketamine alters pulmonary vascular resistance, it would negate the primary reason for the procedure. Because the patient population studied herein resides greater than or equal to 1,200 meters above sea level, concerns about pharmacologic effects on pulmonary vascular resistance are enhanced. Simultaneous pulmonary artery and aortic pressures, thermodilution cardiac outputs, and blood gases were measured in room air (16% oxygen) and with ketamine infusion in 14 patients at cardiac catheterization. Reaction to hypoxia identified 3 groups: normal, intermediate and hyperresponders. The normal responders had normal resistance ratios (0.11) in room air and had little resistance ratio response to hypoxia (+0.02), hyperoxia (-0.03) or ketamine (+0.01). The intermediate responders had a slightly higher but normal resistance ratio (0.20) in room air, and a moderate reaction to hypoxia (+0.13), hyperoxia (-0.08) and ketamine (+0.11). The hyperresponders had an elevated resistance ratio (0.42) in room air and a striking reaction to hypoxia (+0.65), hyperoxia (-0.17) and ketamine (+0.49). Hypoxia and ketamine have a greater effect on resistance ratio than hypoxia alone in patients with reactive pulmonary vascular beds. Ketamine should not be used in children undergoing procedures to establish operability based on pulmonary vascular resistance or pulmonary vascular reactivity.
Altitude↗
Effect of exogenous growth hormone on whole-body and isolated-limb protein kinetics in burned patients.
The effect of growth hormone on protein kinetics was assessed in burned patients during the hyperdynamic phase using N15 lysine and balance data across the leg. Levels of resting energy expenditure and cardiac index were comparably elevated in all patients, but leg blood flow was greater in the patients receiving growth hormone. Growth hormone therapy (0.2 mg/kg per day) significantly stimulated protein synthesis in the whole body and in the studied leg. A hyperinsulinemic clamp, which raised the insulin concentration to more than 1435 pmol/L of blood, caused comparable stimulation of leg protein synthesis in patients not receiving growth hormone, but did not further increase protein synthesis in the growth hormone-treated patients. These results suggest that administration of exogenous growth hormone may limit the peripheral protein wasting in severely injured patients by a mechanism similar to that of insulin.
Adolescent↗
Effect of exogenous growth hormone on glucose utilization in burn patients.
The treatment of burn patients with recombinantly derived human growth hormone (rHGH) appears effective in counteracting protein catabolism. However, exogenous growth hormone is frequently associated with hyperglycemia, an aspect which may limit its usefulness. Therefore, to assess the affect of rHGH on glucose utilization, 13 severely burned patients (65% +/- 4 TBSA burn; mean +/- SEM) began receiving on admission either placebo or rHGH (0.2 mg/kg.d) in a double-blind randomized fashion. While hypermetabolic (percentage REE/predicted REE 1.41 +/- 0.11) fasting oxygen consumption and CO2 production were measured using indirect calorimetry prior to and then during a hyperinsulinemic euglycemic clamp. This experiment demonstrated that rHGH significantly reduced glucose uptake and inhibited glucose oxidation compared to the placebo patients. Since the decreases in glucose oxidation and uptake were proportional, glucose utilization (percentage glucose uptake oxidized) remained similar in both patient groups. Furthermore, the hyperinsulinemic clamp lowered the plasma amino acid concentrations in the control patients while rHGH-treated patients had no significant alterations. In conclusion, exogenous growth hormone therapy induces an insulin resistance in burn patients. Furthermore, since the glucose utilization did not change, it is likely that the mechanism of insulin resistance is due to a deficiency in glucose transport.
Amino Acids↗
Isotopic determination of fibronectin synthesis in humans.
Fibronectin is an opsonic protein that, among other functions, activates the reticuloendothelial system. Accurate measurement of its rate of synthesis is necessary to more fully understand its physiological role in normal and pathological conditions. We have determined the rate of fibronectin synthesis in three normal volunteers using a primed-constant infusion of 15N-glycine and 1,2-13C-leucine, and measuring the incorporation of the isotopes into the protein over 5 days of infusion. In nine additional subjects, the fractional synthetic rate (FSR) of fibronectin was calculated during a 24-hour infusion using urinary hippurate and plasma alpha-ketoisocaproic acid enrichment to represent the precursors for incorporation of labeled glycine and leucine, respectively, into fibronectin. The FSR using glycine and leucine was 1.56 +/- 0.14 and 1.29 +/- 0.04 (%/h), respectively, in the 5-day infusion study, and 1.56 +/- 0.10 versus 1.83 +/- 0.09 (%/h), respectively, in the 24-hour study. The results of the 5-day infusion of 15N-glycine justify the use of urinary hippurate to reflect the precursor enrichment for the determination of the FSR of fibronectin during a shorter (less than 24 hour) infusion period.
Carbon Isotopes↗
Estimating energy requirements in burned children: a new approach derived from measurements of resting energy expenditure.
We examined the determinants of resting energy expenditure (REE) in 127 observations in 56 burned children. Predicted basal energy expenditure (PBEE), body surface area (BSA), and body weight correlated significantly with REE (r2 = 0.76). Days postburn and burn size (% BSA burned) only accounted for 21%, and 24% of the variation in the elevation in REE above PBEE. The single most powerful predictor of REE was PBEE (REE = 1.29 x PBEE); addition of other variables did not improve the prediction. When our recently described activity factor of 1.2 for burn patients is used, the data predict that the average energy requirement to maintain energy balance is 1.55 x PBEE, which is significantly lower than commonly used recommendations, especially for larger burns. The energy required to ensure that 95% of patients achieve energy balance was (1.55 x PBEE) + (2.39 xoff+PBEE0.75), approximately equal to 2 x PBEE. Because the equations presented are derived from measurements of energy expenditure, they represent the most valid approach to estimating energy requirements.
Adolescent↗