NUTRITIONAL STATUS IN CHILDREN WITH CANCER: COMPARISON OF DEUTERIUM OXIDE DILUTION WITH BIOELECTRIC IMPEDANCE ANALYSIS AND ANTHROPOMETRY.

OBJECTIVE: To explore changes in the nutritional status of pediatric cancer patients before and after chemotherapy and evaluate the correlation between deuterium oxide dilution, bioelectric impedance analysis, and anthropometry for assessment of body composition. METHODS: This study included 14 children (aged 5.6 to 13.6 years) and classified them as having hematologic or solid tumors. They had their body composition analyzed according to deuterium oxide, bioelectric impedance, and anthropometric measurements before the first chemotherapy cycle and after three and six months of therapy. RESULTS: The patients in the hematologic tumor group had an increase in weight, height, body mass index, waist, hip, and arm circumference, subscapular skinfold thickness, and fat mass with the isotope dilution technique during chemotherapy. In the solid tumor group, the children showed a reduction in fat-free mass when assessed by bioimpedance analysis. We found a positive correlation between the triceps skinfold thickness and fat mass determined by bioimpedance analysis and deuterium oxide. The arm muscle circumference correlated with the fat-free mass estimated by bioimpedance analysis and deuterium oxide. CONCLUSIONS: Patients with hematologic tumors had an increase in body weight, height, and fat mass, which was not identified in the solid tumor group. The positive correlation between anthropometry (triceps skinfold thickness and arm muscle circumference), deuterium oxide dilution, and bioelectric impedance analysis shows the applicability of anthropometry in clinical practice.

Nutritional status in children with cancer: comparison of deuterium oxide dilution with bioelectric impedance analysis and anthropometry / Estado nutricional em crianças com câncer: comparação entre diluição de deutério, impedância bioelétrica e antropometria

ABSTRACT Objective: To explore changes in the nutritional status of pediatric cancer patients before and after chemotherapy and evaluate the correlation between deuterium oxide dilution, bioelectric impedance analysis, and anthropometry for assessment of body composition. Methods: This study included 14 children (aged 5.6 to 13.6 years) and classified them as having hematologic or solid tumors. They had their body composition analyzed according to deuterium oxide, bioelectric impedance, and anthropometric measurements before the first chemotherapy cycle and after three and six months of therapy. Results: The patients in the hematologic tumor group had an increase in weight, height, body mass index, waist, hip, and arm circumference, subscapular skinfold thickness, and fat mass with the isotope dilution technique during chemotherapy. In the solid tumor group, the children showed a reduction in fat-free mass when assessed by bioimpedance analysis. We found a positive correlation between the triceps skinfold thickness and fat mass determined by bioimpedance analysis and deuterium oxide. The arm muscle circumference correlated with the fat-free mass estimated by bioimpedance analysis and deuterium oxide. Conclusions: Patients with hematologic tumors had an increase in body weight, height, and fat mass, which was not identified in the solid tumor group. The positive correlation between anthropometry (triceps skinfold thickness and arm muscle circumference), deuterium oxide dilution, and bioelectric impedance analysis shows the applicability of anthropometry in clinical practice.

RESUMO Objetivo: Verificar mudanças no estado nutricional de crianças com câncer antes e após o tratamento quimioterápico e avaliar a correlação entre diluição de óxido de deutério, análise de impedância bioelétrica (BIA) e dados antropométricos. Métodos: Quatorze crianças (entre 5,6 e 13,6 anos de idade) foram incluídas e classificadas como tendo tumores hematológicos ou sólidos. A composição corporal foi medida pelo óxido de deutério, impedância bioelétrica e medidas antropométricas antes da primeira quimioterapia e após três e seis meses de terapia. Resultados: Os pacientes do grupo de tumores hematológicos aumentaram o peso, a estatura, o índice de massa corporal, a circunferência da cintura, quadril e braço, dobra cutânea subescapular e a massa gorda com a técnica de diluição isotópica durante o tratamento quimioterápico. No grupo de tumores sólidos, as crianças mostraram uma redução na massa magra quando avaliadas por análise de impedância. Houve uma correlação positiva entre a dobra cutânea tricipital e a massa gorda determinadas pela análise de impedância e pelo óxido de deutério. A circunferência muscular do braço correlacionou-se com a massa magra estimada pela análise de impedância e pelo óxido de deutério. Conclusões: Pacientes com tumores hematológicos tiveram aumento no peso corporal, estatura e massa gorda, o que não foi observado naqueles com tumores sólidos. A boa correlação entre a antropometria (dobra cutânea tricipital e circunferência muscular do braço), a diluição do óxido de deutério e a análise da impedância bioelétrica mostra a aplicabilidade da antropometria na prática clínica.

Measurement of Leukemic B-Cell Growth Kinetics in Patients with Chronic Lymphocytic Leukemia.

Cell proliferation plays a central role in the pathogenesis of every neoplastic disease as well as many other types of illness. Labeling of newly replicated DNA with deuterium (2H), a nonradioactive isotope of hydrogen, administered to the patients in drinking water (2H2O) is a safe and reliable method to measure the in vivo birth rates of cells. Here, we describe a protocol to measure chronic lymphocytic leukemia B-cell birth/proliferation and death rates over time using this approach.

Determination of low levels of 2H-labeling using high-resolution mass spectrometry: application in studies of lipid flux and beyond.

RATIONALE: The ability to measure low levels of (2)H-labeling is important in studies of metabolic flux, e.g. one can estimate lipid synthesis by administering (2)H2O and then measuring the incorporation of (2)H into fatty acids. Unfortunately, the analyses are complicated by the presence of more abundant naturally occurring stable isotopes, e.g. (13)C. Conventional approaches rely on coupling gas chromatographic separation of lipids with either quadrupole-mass spectrometry (q-MS) and/or pyrolysis-isotope ratio mass spectrometry (IRMS). The former is limited by high background labeling (primarily from (13)C) whereas the latter is not suitable for routine high-throughput analyses. METHODS: We have contrasted the use of continuous flow-pyrolysis-IRMS against high-resolution mass spectrometry (i.e. Qq-FT-ICR MS) for measuring the (2)H-enrichment of fatty acids and peptides. RESULTS: In contrast to IRMS, which requires ~30 min per analysis, it is possible to measure the (2)H-enrichment of palmitate via direct infusion high-resolution mass spectrometry (HRMS) in ~3 min per sample. In addition, Qq-FT-ICR MS enabled measurements of the (2)H-enrichment of peptides (which is not possible using IRMS). CONCLUSIONS: High-resolution mass spectrometry can be used to measure low levels of (2)H-labeling so we expect that this approach will enhance studies of metabolic flux that rely on (2)H-labeled tracers, e.g. (2)H2O. However, since the high-resolution analyses require greater amounts of a given analyte one potential limitation centers on the overall sensitivity. Presumably, future advances can overcome this barrier.

Hydrogen and N-acetyl-L-cysteine rescue oxidative stress-induced angiogenesis in a mouse corneal alkali-burn model.

PURPOSE: To investigate the role of reactive oxygen species (ROS) as the prime initiators of the angiogenic response after alkali injury of the cornea and observe the effects of antioxidants in preventing angiogenesis. METHODS: The corneal epithelia of SOD-1-deficient mice or wild-type (WT) mice were removed after application of 0.15 N NaOH to establish the animal model of alkali burn. ROS production was semiquantitatively measured by dihydroethidium (DHE) fluorescence. Angiogenesis was visualized by CD31 immunohistochemistry. The effects of the specific NF-κB inhibitor DHMEQ, the antioxidant N-acetyl-L-cysteine (NAC), and hydrogen (H2) solution were observed. RESULTS: ROS production in the cornea was enhanced immediately after alkali injury, as shown by increased DHE fluorescence (P<0.01). NF-κB activation and the upregulation of vascular endothelial growth factor (VEGF) and monocyte chemoattractant protein-1 (MCP-1) were significantly enhanced (P<0.01), leading to a significantly larger area of angiogenesis. Angiogenesis in SOD-1-/- mice corneas were significantly higher in WT mice (P<0.01), confirming the role of ROS. Pretreatment with the specific NF-κB inhibitor DHMEQ or the antioxidant NAC significantly reduced corneal angiogenesis by downregulating the NF-κB pathway (P<0.01) in both WT and SOD-1-/- mice. Furthermore, we showed that irrigation of the cornea with hydrogen (H2) solution significantly reduced angiogenesis after alkali-burn injury (P<0.01). CONCLUSIONS: Immediate antioxidant therapy with H2-enriched irrigation solution is a new potent treatment of angiogenesis in cornea to prevent blindness caused by alkali burn.

Isotopologue distributions of peptide product ions by tandem mass spectrometry: quantitation of low levels of deuterium incorporation.

Protonated molecular peptide ions and their product ions generated by tandem mass spectrometry appear as isotopologue clusters due to the natural isotopic variations of carbon, hydrogen, nitrogen, oxygen, and sulfur. Quantitation of the isotopic composition of peptides can be employed in experiments involving isotope effects, isotope exchange, and isotopic labeling by chemical reactions and in studies of metabolism by stable isotope incorporation. Both ion trap and quadrupole-time of flight mass spectrometry are shown to be capable of determining the isotopic composition of peptide product ions obtained by tandem mass spectrometry with both precision and accuracy. Tandem mass spectra of clusters of isotopologue ions obtained in profile mode are fit by nonlinear least squares to a series of Gaussian peaks which quantify the Mn/M0 values which define the isotopologue distribution (ID). To determine the isotopic composition of product ions from their ID, a new algorithm that predicts the Mn/M0 ratios and obviates the need to determine the intensity of all of the ions of an ID is developed. Consequently a precise and accurate determination of the isotopic composition of a product ion may be obtained from only the initial values of the ID, however, the entire isotopologue cluster must be isolated prior to fragmentation. Following optimization of the molecular ion isolation width, fragmentation energy, and detector sensitivity, the presence of isotopic excess (2H, 13C, 15N, 18O) is readily determined within 1%. The ability to determine the isotopic composition of sequential product ions permits the isotopic composition of individual amino acid residues in the precursor ion to be determined.

Measuring gluconeogenesis using a low dose of 2H2O: advantage of isotope fractionation during gas chromatography.

The contribution of gluconeogenesis to glucose production can be measured by enriching body water with (2)H(2)O to approximately 0.5% (2)H and determining the ratio of (2)H that is bound to carbon-5 vs. carbon-2 of blood glucose. This labeling ratio can be measured using gas chromatography-mass spectrometry after the corresponding glucose carbons are converted to formaldehyde and then to hexamethylenetetramine (HMT). We present a technique for integrating ion chromatograms that allows one to use only 0.05% (2)H in body water (i.e., 10 times less than the current dose). This technique takes advantage of the difference in gas chromatographic retention times of naturally labeled HMT and [(2)H]HMT. We discuss the advantage(s) of using a low dose of (2)H(2)O to quantify the contribution of gluconeogenesis.

Non-invasive tumour blood perfusion measurement by 2H magnetic resonance.

Deuterium uptake into foot-implanted C3H murine mammary carcinomas was measured non-invasively by 2H NMR spectroscopy at 46 MHz after i.v. injection. The arterial input function (AIF) was estimated from 2H NMR measurements with a second radiofrequency coil externally located over the heart. Tumour and heart data were acquired over the same time period by means of a switch automatically activated every 1.6-3.2 s. Although the AIF data were, in general, partly contaminated by signals from adjacent tissue, a mathematical fitting procedure involving simultaneous fitting of the AIF and the tumour kinetics gave robust results for tumour blood perfusion (TBP): up to four repeat TBP measurements were made in 14 out of 20 untreated animals and TBP could be measured before and after treatment in 14 out of 15 animals. The ability of this technique to measure changes in blood perfusion was assessed using hydralazine, which decreased TBP from 91 to 29 ml 100 g(-1) min(-1) and this was comparable to a 70% reduction in relative TBP measured by laser Doppler flowmetry.

Evaluación del consumo de leche humana por dilución con deuterio y detección por espectroscopia de infrarrojo / Assessment of human milk consumption by dilution with deuterium and detection by infrared spectroscopy

Objetivo. Adaptar la metodología de evaluación de ingestión de leche humana con isótopos estables utilizando un espectroscopio infrarrojo en lugar de un espectrómetro de masas. Material y métodos. Previo consentimiento informado, 12 mujeres que estaban amamantando recibieron 30 g de D2O por vía oral. a las 4 horas y 1, 3, 6, 9 y 14 días depués de la dosificación, se les tomó una muestra de saliva. Las muestras de saliva de los bebés se tomaron a los 1,2,5,6,13 y 14 días. Todas las muestras fueron preparadas por sublimación y centrifucación y la determinación de la concentración de D2O se determinó por espectroscopía de infrarrojo. Resultados. La recuperación del sistema de sublimación fue superior al 95 por ciento del volumen total tanto de agua sola como de saliva. Se encontró que con tres evaluaciones se alcanza la misma confiabilidad que con 5 evaluaciones. Los volúmenes de ingestión de leche humana fueron similares a los informados por otros autores

Total energy expenditure and physical activity as assessed by the doubly labeled water method in Swedish adolescents in whom energy intake was underestimated by 7-d diet records.

Swedish children and adolescents may be adopting a sedentary lifestyle with low energy expenditures and intakes, but no quantitative data are available. The purpose of the present study in 50 adolescents aged 15 y was to investigate whether assessment of total energy expenditure (TEE) and physical activity level (PAL) by the doubly labeled water method and indirect calorimetry and estimation of energy intake by a 7-d diet record would indicate physical inactivity. The boys' (n = 25) mean weight was 112% and the girls' (n = 25) was 109% of Swedish reference values from 1976; the mean height of both boys and girls was 102% of those reference values. Mean TEE in the boys and girls, 13.82 +/- 1.90 and 10.70 +/- 1.59 MJ/d, and mean PAL (TEE/basal metabolic rate), 1.89 +/- 0.16 and 1.79 +/- 0.22, respectively, were nonsignificantly higher than corresponding figures from other published studies. Mean energy intake as a percentage of TEE was 81.9 +/- 17.9% in the boys and 78.3 +/- 16.4% in the girls. Significant negative correlations were found both between energy intake as a percentage of TEE and percentage body fat and between energy intake as a percentage of TEE and body mass index. These results add to the evidence that 7-d diet records underestimate energy intake in adolescents, particularly those with a tendency for overweight and increased body fat. The results support indications of a trend of increasing body weight and height in Swedish adolescents, but conflict with the presumptions of low physical activity, low energy expenditure, and low energy intake. These results support the view that current recommendations for energy intake during adolescence are too low.