Metabolism of parenterally administered fat emulsions in the rat: studies of fatty acid oxidation with 1-13C- and 8-13C-labelled triolein.

To reassess the hypothesis that fatty acid catabolism occurs to completion via beta-oxidation, male Sprague-Dawley rats receiving continuous total parenteral nutrition (TPN) including 43% energy as fat were infused with [1-(13)C]- or [8-(13)C]triolein. Expired CO2 was collected continuously for 4 h and its 13C:12C ratio determined by isotope-ratio mass spectrometry. Bicarbonate retention was also assessed over 4 h by infusion of NaH14CO3 and measurement of the expired 14CO2. A possible loss of label from [8-(13)C]oleic acid from the citric acid cycle via labelled acetyl-CoA without oxidation to CO2 was assessed by infusing further animals with acetate labelled with 14C either at C atoms 1 or 2 and determination of its conversion to expired 14CO2. At isotopic steady state, 63.2 (SE 1.6)% (n 8) of the infused [1-(14)C]acetate and 46.0 (SE 1.2)% (n 8) of [2-(14)C]acetate was recovered as expired 14CO2. After correction for bicarbonate retention and non-oxidative isotope loss, 37.3 (SE 1.2)% (n 20) of the [1-(13)C]triolein was found to have been oxidized, whereas 32.6 (SE 1.0)% (n 20) of the [8-(13)C]triolein was oxidized (P < or = 0.01). The lower oxidation of the C atom at position 8 of oleic acid than that at position 1 indicates incomplete oxidative breakdown of the fatty acid after entering beta-oxidation.

Assessment of whole body L-leucine oxidation by noninvasive L-[1-13C]leucine breath tests: a reappraisal in patients with maple syrup urine disease, obligate heterozygotes, and healthy subjects.

Suitability of a recently proposed noninvasive L-[13C]leucine breath test for assessment of whole body leucine oxidation in maple syrup urine disease (MSUD) was examined. Oral L-[1-13C]leucine loads (38 micromol/kg body weight) were performed in overnight fasted MSUD patients (n = 6, classical form), obligate heterozygote parents (n = 6), and control subjects (n = 10). Three-hour 13CO2 exhalation kinetics were evaluated using curve fitting procedures. Venous blood was obtained in most cases and analyzed for 13C-labeled plasma metabolites. In control subjects, maximal 13CO2 exhalation was reached at tmax = 55 +/- 18 min. Cumulative 13CO2 output at 3 h amounted to 4.7 +/- 0.7 micromol x (kg body weight)(-1). Estimated total 3CO2 exhalation was 7.2 +/- 1.4 micromol x (kg body weight)(-1) (19.0 +/- 3.6% of the dose). Half of this amount was expired at t1/2 = 130 +/- 18 min. The data show a considerable degree of intersubject variability. Intraindividual variability was comparable, however, when checked in two volunteers. In obligate heterozygotes, 13CO2 kinetics were similar to controls (tmax = 35 +/- 8 min, t1/2 = 95 +/- 16 min). Total 13CO2 output [5.7 +/- 1.4 micromol x (kg body weight)(-1)] tended to be in the lower control range. None of the MSUD patients under study exhibited a significant increase in 13CO2 output after load. Maximal increase of label in plasma 4-methyl-2-oxopentanoate, the physiologic precursor of 13CO2, was 16.1 +/- 3.5 MPE in control subjects. In MSUD, label dilution was increased and correlated with the patients' leucine/4-methyl-2-oxopentanoate plasma levels. Considering the generally high variability of 13CO2 output and the unstable substrate pools in MSUD, we discuss the limitations of whole body leucine oxidation measurements by noninvasive approaches.

Prevalence of Helicobacter pylori infection in Nicaraguan children with persistent diarrhea, diagnosed by the 13C-urea breath test.

BACKGROUND: The impairment of gastric acid barrier caused by Helicobacter pylori (H. pylori) at the onset of infection may predispose to small bowel bacterial overgrowth, which could contribute to persistent diarrhea. METHODS: Using the 13C-urea breath test, we determined the prevalence of H. pylori infection in 123 Nicaraguan children from Tipitapa, aged 1 to 65 months, from a low socioeconomic background. RESULTS: The overall prevalence of H. pylori infection was 77.2% (95/123). The prevalence varied with age and was significantly (p < 0.001) higher in infants < or = 12 months than in children aged 13-65 months, 91% (57/63) as against 63% (38/60). H. pylori infection was present in 44 of 59 (75%) children suffering from persistent diarrhea compared with 51 of 64 (80%) age-matched asymptomatic controls. In the diarrheal group, 20 of 59 (34%) children presented with malnutrition, and 16 (80%) of them showed H. pylori infection. In the control group, 20 of 64 (31%) were malnourished, and 14 (70%) of them showed H. pylori infection. CONCLUSIONS: In Nicaragua, H. pylori is acquired in early infancy. The high prevalence among children in the first 12 months of life and the lower infection rate between 1 and 5 years of age suggest a loss or clearance of infection, also an occasional finding in adults. H. pylori infection appears to be not a risk factor for persistent diarrhea or malnutrition in Nicaraguan children.

Application of isotope-selective nondispersive infrared spectrometry (IRIS) for evaluation of [13C]octanoic acid gastric-emptying breath tests: comparison with isotope ratio-mass spectrometry (IRMS).

Suitability of isotope-selective nondispersive infrared spectrometry (IRIS) for evaluation of [13C]octanoic acid gastric-emptying breath test was assessed and compared with standard isotope ratio-mass spectrometry (IRMS). The estimated bias of IRMS and IRIS measurements of baseline-corrected 13CO2 exhalation amounted to +/-0.1 and +/-0.6 delta delta values (n = 360), respectively. In breath tests performed on 60 diabetic patients, the gastric emptying parameters were calculated by nonlinear regression analysis of the time course of 13CO2 exhalation: half-emptying time (t1/2,breath, 90 +/- 39 min), lag phase (tlag,breath, 34 +/- 27 min), and gastric emptying coefficient (GEC, 2.9 +/- 0.5). A reasonable linear correlation was found between the two methods (y = IRIS, x = IRMS) with respect to delta delta values (y = 0.35 + 0.92x, r = 0.985, Sy[symbol: see text]x = +/-0.6, n = 1116) and a rather good agreement of the computed gastric emptying parameters was obtained (t1/2,breath: y = 0.99x + 4.06, Sy[symbol: see text]x = +/- 6.3; tlag, breath: y = 0.97x + 0.96, Sy[symbol: see text]x = +/-3.4; GEC: y = 0.97x - 0.01, Sy[symbol: see text]x = +/-0.09).

[13C]octanoic acid breath test for non-invasive assessment of gastric emptying in diabetic patients: validation and relationship to gastric symptoms and cardiovascular autonomic function.

Since there is a need for a widely applicable non-invasive test to assess gastric emptying in diabetic patients, we evaluated the sensitivity, specificity, and reproducibility of the [13C]octanoic acid breath test as compared with scintigraphy. Moreover, we examined the relationship between the breath test indices and gastric symptoms, cardiovascular autonomic function, and metabolic parameters. Forty healthy control subjects and 34 diabetic patients were studied. Three indices of gastric emptying, assessed by the breath test, were computed: half-emptying time (t1/2breath), gastric emptying coefficient (GEC), and lag phase. Furthermore, the half-emptying time, measured by scintigraphy (t1/2scint), was calculated and gastric symptoms and cardiovascular autonomic neuropathy (CAN) were scored. The coefficients of variation of day-to-day reproducibility in 10 healthy subjects were 29.6% for t1/2breath, 7.4% for GEC, and 46.5% for lag phase. An abnormal delay for t1/2scint (> 100 min) or t1/2breath (> 200 min) was noted in 12 patients. Based on the results for t1/2scint, the sensitivity of t1/2breath and GEC was 75% and the specificity was 86%. Both t1/2breath (rs = 0.523; p < 0.05) and GEC (r2 = -0.594; p < 0.05) were significantly associated with the gastric symptom score. A significant relationship to the CAN score was demonstrated for t1/2breath (rs = 0.448; p < 0.05), GEC (rs = -0.467; p < 0.05), and t1/2scint (rs = 0.602; p < 0.05). There were no significant associations of the breath test indices with the blood glucose levels during the test, HbA1c, age, and duration of diabetes. In patients with abnormal t1/2scint (n = 12) not only was t1/2breath significantly prolonged and GEC reduced, but also the scores of CAN and gastric symptoms were significantly increased as compared with those who had a normal t1/2scint (n = 22). We conclude that the [13C]octanoic acid breath test represents a suitable measure of delayed gastric emptying in diabetic patients which is associated with the severity of gastric symptoms and CAN but not affected by the blood glucose level.

Utilisation of amino acid mixtures in adolescents with phenylketonuria.

The nutritional regimen of patients with phenylketonuria (PKU) comprises a diet of natural proteins and phenylalanine (Phe)-free amino acid (AA) mixture. The main daily protein requirement is covered by a Phe-free AA mixture. In an adult with PKU, the consumption of the daily AA requirement in one single dose at breakfast caused nausea and vomiting. Therefore, four studies were designed to investigate the adverse and metabolic effects resulting from large intakes of AA mixtures used in the treatment of PKU patients with respect to the following: (1) biochemical effects following consumption of one single dose of Phe-free AA mixture in healthy persons; (2) transient metabolic changes caused by different individual regimens of AA intake in healthy persons and in one PKU patient; (3) nitrogen excretion in PKU patients taking the AA mixture in two or three portions; and (4) catabolic metabolism of AA in a PKU patient. In healthy subjects following the ingestion of the AA mixture in one bolus there was an increase in the blood levels of the given AA and also an increase in blood insulin concentration and a decrease in blood glucose concentration. These changes were less marked when the AA mixture was divided into three portions per day. In contrast, in a PKU patient following the ingestion of AA there was an increase in blood glucose. The urinary nitrogen excretion was greater in PKU patients when one compared to three portions of AA mixture was taken. The consumption of the daily requirement of AA mixture in one single does produced increased catabolism in a PKU patient. In conclusion it is recommended that the total daily amount of AA mixture should be divided into a minimum of three portions.

Specific and dose-dependent enzyme induction by omeprazole in human beings.

Omeprazole induces hepatic cytochrome P-4501A2. In a previous study this effect was shown to be significant in vivo in 6 poor metabolizers, including 1 intermediate metabolizer, but not in 12 extensive metabolizers of S-mephenytoin after 7 days of treatment with 40 mg/day omeprazole. In this study, the specificity of the inducing potential of omeprazole was investigated in these volunteers. Furthermore, in eight of the extensive metabolizers the dose-dependence of cytochrome P-450 1A2 induction was evaluated. Cytochrome P-450 1A2 activity was monitored by means of the 13C-[N3-methyl]caffeine breath test and by means of plasma caffeine clearance before omeprazole treatment with 120 mg/day, on the seventh day of dosage and after a 7-day washout. Omeprazole plasma concentration was measured. Results were compared with those after 40 mg. gamma-Glutamyltransferase activity in serum, as well as urinary excretion of D-glucaric acid and 6 beta-hydroxycortisol, were measured on the same study days in all study groups (n = 26). In the eight extensive metabolizers the breath test indicated a dose-dependent increase of cytochrome P-450 1A2 activity of 8.5% +/- 15.0% (40 mg, mean +/- SD, NS) and 27.2% +/- 16.5% (120 mg, p = 0.002). Caffeine clearance was increased by 31.6% +/- 20.7% (p < 0.001) with the higher dose. None of the study groups exhibited a significant increase of gamma-glutamyltransferase activity or urinary excretion of D-glucaric acid or 6 beta-hydroxycortisol. This was in contrast to the phenobarbital-type induction observed after treatment with antiepileptic drugs. Induction by omeprazole seems to be restricted to cytochrome P-450 1A enzymes.(ABSTRACT TRUNCATED AT 250 WORDS)

Dependence of the utilization of a phenylalanine-free amino acid mixture on different amounts of single dose ingested. A case report.

For patients with phenylketonuria the daily ingested phenylalanine-free amino acid mixture is the most important source of nitrogen. It is recommended to ingest one third of the total amount combined with main meals. Some patients, especially the older ones, do not follow this recommendation; they ingest the entire daily amount of amino acid mixture in one portion. This intake mode leads to an increased oxidative utilization of the amino acids. To set up an example for this metabolic phenomenon, a 13C-leucine breath test was performed in one female phenylketonuric patient. She ingested a third of her daily amount of the amino acid mixture combined with an oral tracer of 3 mg 13C-leucine/kg body weight at breakfast. The breath test was carried out by a standardized time schedule over 5 h. Three days later the breath test was repeated when she ingested the total amount of amino acid mixture in only one portion at breakfast. Total daily caloric intake and food composition were not changed. On both days a 24 h urine was collected to determine total nitrogen loss. The 13C-content of expired air was analysed by gas isotope ratio mass spectrometry, the total nitrogen content was determined using a combustion unit. The 13C-elimination rate as a percentage of the applied 13C-tracer was 9.5% on the first test day as compared to 19.6% on the 2nd day. The corresponding total nitrogen excretion was increased (4.3-6.9 g/24 h).(ABSTRACT TRUNCATED AT 250 WORDS)

Increase of cytochrome P450IA2 activity by omeprazole: evidence by the 13C-[N-3-methyl]-caffeine breath test in poor and extensive metabolizers of S-mephenytoin.

Omeprazole has been shown to induce cytochrome P450IA1 and P450IA2 activity in vitro. To reflect cytochrome P450IA2 (CYP1A2) activity in vivo, the 13C-[N-3-methyl]-caffeine breath test was conducted in 18 volunteers: 12 extensive metabolizers, one intermediate metabolizer, and five poor metabolizers of S-mephenytoin. Breath tests were performed before treatment with an oral dose of 40 mg omeprazole, on the seventh day of treatment, and after a 7-day washout period. The mean percentage exhalation of the 13C test dose, as determined by 13CO2 in breath during 8 hours, was 23.0% +/- 8.0% (n = 18) before treatment. The largest increases in exhalation rate of 13CO2 were observed in the poor metabolizers and the intermediate metabolizers (range, 12.8% to 62.9%; median, 38.9%); median area under the plasma concentration-time curves (AUC) of omeprazole was four times higher than in the extensive metabolizers. The change after omeprazole treatment in extensive metabolizers ranged from -9.8% to +47.7% (median, 12.3%; n = 12) of pretreatment values. In both groups exhalation rates of 13CO2 returned to near pretreatment values within the 7-day washout period (24.2% +/- 7.8%; n = 17). Changes in the 13C-caffeine breath test correlated well with both the pretreatment value (R = -0.67, p = 0.003; n = 18) and the plasma AUC of omeprazole (R = 0.61, p = 0.007; n = 18). Therapeutic doses of omeprazole seem to induce CYP1A2 activity in poor metabolizers, whereas they exert minor inducing effects in extensive metabolizers of S-mephenytoin.

[Phenylalanine-free amino acid mixture: metabolic effects in relation to a single dose]. / Phenylalaninfreie Aminosäurenmischung: Stoffwechselwirkung in Abhängigkeit von der Einzeldosis.

In the dietetic treatment of phenylketonuric patients phenylalanine free amino acid mixtures are given to meet the protein requirement of the patient. In four healthy probands we tested the metabolic effects of four different PKU-preparations given in variable amounts ingested with carbohydrate and fat containing meals. We determined glucose, insulin, amino acids, and urea in blood. Following the amino acid load we saw an increased insulin output with a hypoglycemic reaction or, with smaller amounts a lack of the normal postprandial blood glucose increase. Hyperaminoacidemias depended on the amount of amino acids ingested. The increase of blood urea found suggests that part of the amino acids were degraded. To compare the results we studied a PKU-patient. He showed corresponding but milder effects.

[Effect of carbohydrate administration on leucine oxidation in newborn infants. Studies using the 13C leucine breath test]. / Einfluss der Kohlenhydratzufuhr auf die Leuzinoxydation bei Neugeborenen. Untersuchungen mit dem 13C-Leuzin-Atemtest.

The branched-chain amino acid leucine plays an important role in the protein metabolism of human beings. It not only inhibits protein degradation but also stimulates protein synthesis. The oxidation rate of leucine and the influence which nutritional conditions have on this amino acid can be measured with the intravenous 13C-leucine breath test. In order the apply the breath test on newborn infants, the required dosage of L-(1-13C)-leucine and the reproducibility of the test had, firstly, to be determined. Following this, the extent to which the leucine oxidation rate was influenced by a simultaneous carbohydrate intake was investigated. An evident discrimination between the 13CO2-exhalation and the 13CO2-baseline exhalation is demonstrated after a bolus injection of 1 mg L-(1-13C)-leucine/kg B.W. We were able to measure reproducible values of the leucine oxidation rate in newborn infants with a tracer dosage of 4 mg L-(1-13C)-leucine/kg B.W. We found that a higher intake of carbohydrate given at the same time produced a lower rate of leucine oxidation, which indicates increased utilization of leucine for the benefit of protein synthesis.

Impaired fat utilization in parenterally fed low-birth-weight infants suffering from sepsis.

Lipid infusion in low-birth-weight infants suffering from sepsis is still controversial. Consequently, we investigated the fat tolerance in six low-birth-weight infants with sepsis and 15 low-birth-weight infants without sepsis. For measurement of fat clearance, we assayed the serum concentrations of triglycerides enzymatically, and of the free fatty acids by colorimetric micromethod. The fatty acid oxidation was analyzed with the [13C]triolein breath test by means of ratio-mass spectrometry. The infants were maintained on continuous parenteral nutrition with various amounts of soybean oil emulsion (1 g, 2 g, and 3 g fat/kg body weight per day). Comparing the lipid infusion of 1 and 2 g fat/kg body weight per day between the two groups, we found triglyceride and free fatty acid values in both groups to be in the normal range. At a dose of 3 g of fat/kg body weight per day, septic low-birth-weight infants showed a significantly higher concentration of triglycerides (2.02 +/- 0.46 mmol/liter) and of free fatty acids (2.06 +/- 0.45 mmol/liter) than the nonseptic low-birth-weight infants (triglycerides: 1.09 +/- 0.43 mmol/liter; free fatty acids: 1.05 +/- 0.41 mmol/liter). The low-birth-weight infants with sepsis showed a reduced fat oxidation rate of 16.0 +/- 1.5% in contrast to that of the low-birth-weight infants without sepsis, whose rate was 38.4 +/- 1.8%. Accordingly, we apply dosages not exceeding 2 g of fat/kg body weight per day to septic low-birth-weight infants.

[Fat utilization in newborn infants with and without heparin administration. Comparative study with the 13C-triolein breath test]. / Fettutilisation bei Frühgeborenen mit und ohne Heparingabe. Vergleichende Untersuchung mit dem 13C-Triolein-Atemtest.

The elimination of parenterally administered lipids from the bloodstream of premature infants can be accelerated by activation of the lipoprotein-lipase using heparin. We have no evidence that the free fatty acids increasing under enhanced lipolytical activity are utilized for energy production. For this reason, the oxidation rates of intravenously administered lipids in premature infants are examined both with and without heparin. Triolein marked with 13C and processed in soybean oil is administered intravenously at a dosage of 10 mg/kg. 13CO2 results from fatty acid oxidation and is exhaled through the lungs, whereafter it is collected in separate breath samples over a period of 6 hours and determined by mass spectrometry. The examination was performed in 5 premature infants, first without heparin, then after heparin injection (10 U/kg). The extent of 13CO2 exhalation was not significantly influenced by heparin. Without heparin supply we measured a fatty acid oxidation of 32.0 +/- 2.57% which was the same (31.6 +/- 2.34%) after heparin injection. Single intravenous administration of 10 U heparin/kg does not cause increased fatty acid oxidation in premature infants.