Precursors of hexoneogenesis within the human mammary gland.

The human mammary gland is capable of de novo synthesis of glucose and galactose (hexoneogenesis); however, the carbon source is incompletely understood. In this study, we investigated the role of acetate, glutamine, lactate and glycerol as potential carbon sources for hexoneogenesis. Healthy breastfeeding women were studied following a 24-h fast on two occasions separated by 1-3 wk. Five women were infused with [U-¹³C]lactate or [1,2-¹³C2]glutamine and five women with [U-¹³C]glycerol or [1,2-¹³C2]acetate. Enrichments of ¹³C in plasma and milk substrates were analyzed using GC-MS. Infusion of labeled lactate, glycerol, glutamine, and acetate resulted in plasma glucose being 22.0±3.7, 11.2±1.0, 2.5±0.5, and 1.3±0.2% labeled, respectively. Lactate, glutamine, or acetate did not contribute to milk glucose or galactose (0-2%). In milk, ¹³C-free glycerol enrichment was one-fourth that in plasma but free glycerol concentration in milk was fourfold higher than in plasma. Using [U-¹³C]glycerol and by accounting for tracer dilution, glycerol alone contributed to 10±2 and 69±11% of the hexoneogenesis of milk glucose and galactose, respectively. During [U-¹³C]glycerol infusion, the ratio of M3 enrichment on 4-6 carbons/M3 on 1-3 carbons of galactose was higher (P<0.05, 1.22±0.05) than those of glucose in plasma (1.05±0.03) and milk (1.07±0.02). Reanalysis of samples from a previous study involving [U-¹³C]glucose infusion alone suggested labeling a portion of galactose consistent with pentose phosphate pathway (PPP) activity. We conclude that, although lactate contributed significantly to gluconeogenesis, glycerol alone provides the vast majority of substrate for hexoneogenesis. The relative contribution of the PPP vs. the reversal Embden-Meyerhof pathway to hexoneogenesis within the human mammary gland remains to be determined.

De novo synthesis of milk triglycerides in humans.

UNLABELLED: Mammary gland (MG) de novo lipogenesis contributes significantly to milk fat in animals but little is known in humans. OBJECTIVE: To test the hypothesis that the incorporation of (13)C carbons from [U-(13)C]glucose into fatty acids (FA) and glycerol in triglycerides (TG) will be greater: 1) in milk than plasma TG, 2) during a high-carbohydrate (H-CHO) diet than high-fat (H-FAT) diet, and 3) during feeding than fasting. Seven healthy, lactating women were studied on two isocaloric, isonitrogenous diets. On one occasion, subjects received diets containing H-FAT or H-CHO diet for 1 wk. Incorporation of (13)C from infused [U-(13)C]glucose into FA and glycerol was measured using GC-MS and gene expression in RNA isolated from milk fat globule using microarrays. Incorporation of (13)C2 into milk FA increased with increased FA chain length from C2:0 to C12:0 but progressively declined in C14:0 and C16:0 and was not detected in FA>C16. During feeding, regardless of diets, enrichment of (13)C2 in milk FA and (13)C3 in milk glycerol were ∼ 3- and ∼ 7-fold higher compared with plasma FA and glycerol, respectively. Following an overnight fast during H-CHO and H-FAT diets, 25 and 6%, respectively, of medium-chain FA (MCFA, C6-C12) in milk were derived from glucose but increased to 75 and 25% with feeding. Expression of genes involved in FA or glycerol synthesis was unchanged regardless of diet or fast/fed conditions. The human MG is capable of de novo lipogenesis of primarily MCFA and glycerol, which is influenced by the macronutrient composition of the maternal diet.

Effect of ghrelin on glucose regulation in mice.

Improvement of glucose metabolism after bariatric surgery appears to be from the composite effect of the alterations in multiple circulating gut hormone concentrations. However, their individual effect on glucose metabolism during different conditions is not clear. The objective of this study was to determine whether ghrelin has an impact on glycogenolysis, gluconeogenesis, and insulin sensitivity (using a mice model). Rate of appearance of glucose, glycogenolysis, and gluconeogenesis were measured in wild-type (WT), ghrelin knockout (ghrelin(-/-)), and growth hormone secretagogue receptor knockout (Ghsr(-/-)) mice in the postabsorptive state. The physiological nature of the fasting condition was ascertained by a short-term fast commenced immediately at the end of the dark cycle. Concentrations of glucose and insulin were measured, and insulin resistance and hepatic insulin sensitivity were calculated. Glucose concentrations were not different among the groups during the food-deprived period. However, plasma insulin concentrations were lower in the ghrelin(-/-) and Ghsr(-/-) than WT mice. The rates of gluconeogenesis, glycogenolysis, and indexes of insulin sensitivity were higher in the ghrelin(-/-) and Ghsr(-/-) than WT mice during the postabsorptive state. Insulin receptor substrate 1 and glucose transporter 2 gene expressions in hepatic tissues of the ghrelin(-/-) and Ghsr(-/-) were higher compared with that in WT mice. This study demonstrates that gluconeogenesis and glycogenolysis are increased and insulin sensitivity is improved by the ablation of the ghrelin or growth hormone secretagogue receptor in mice.

Gluconeogenesis is not regulated by either glucose or insulin in extremely low birth weight infants receiving total parenteral nutrition.

OBJECTIVE: To determine potential factors regulating gluconeogenesis (GNG) in extremely low birth weight infants receiving total parenteral nutrition. STUDY DESIGN: Seven infants (birth weight, 0.824 ± 0.068 kg; gestational age, 25.4 ± 0.5 weeks; postnatal age, 3.3 ± 0.2 days) were studied for 11 hours, with parenteral lipid and amino acid therapy continued at prestudy rates. Glucose was supplied at prestudy rates for the first 5 hours (period 1) and was then reduced to 6 mg/kg·min for 1 hour and further to ~3 mg/kg·min for 5 hours (period 2). A total of 2.5 mg/kg·min of the glucose was replaced by [U-(13)C]glucose throughout the study for measurements of glucose production and GNG. Concentrations of glucose, insulin, glucagons, and cortisol were determined. RESULTS: GNG and glucose production remained unchanged (2.12 ± 0.23 vs. 1.84 ± 0.25 mg/kg·min [P = NS] and 2.44 ± 0.27 vs. 2.51 ± 0.31 mg/kg·min [P = NS], respectively), despite a 60% reduction of the glucose infusion rate and subsequent 30% (124.7 ± 10.8 to 82.6 ± 8.9 mg/dL; P = .009) and 70% (26.9 ± 4.7 to 6.6 ± 0.4 µU/mL; P = .002) decreases in glucose and insulin concentrations, respectively. Cortisol and glucagon concentrations remained unchanged. CONCLUSION: In extremely low birth weight infants receiving total parenteral nutrition, GNG is a continuous process that is not affected by infusion rates of glucose or concentrations of glucose or insulin.

Gene expression in the human mammary epithelium during lactation: the milk fat globule transcriptome.

The molecular physiology underlying human milk production is largely unknown because of limitations in obtaining tissue samples. Determining gene expression in normal lactating women would be a potential step toward understanding why some women struggle with or fail at breastfeeding their infants. Recently, we demonstrated the utility of RNA obtained from breast milk fat globule (MFG) to detect mammary epithelial cell (MEC)-specific gene expression. We used MFG RNA to determine the gene expression profile of human MEC during lactation. Microarray studies were performed using Human Ref-8 BeadChip arrays (Illumina). MFG RNA was collected every 3 h for 24 h from five healthy, exclusively breastfeeding women. We determined that 14,070 transcripts were expressed and represented the MFG transcriptome. According to GeneSpring GX 9, 156 ontology terms were enriched (corrected P < 0.05), which include cellular (n = 3,379 genes) and metabolic (n = 2,656) processes as the most significantly enriched biological process terms. The top networks and pathways were associated primarily with cellular activities most likely involved with milk synthesis. Multiple sampling over 24 h enabled us to demonstrate core circadian clock gene expression and the periodicity of 1,029 genes (7%) enriched for molecular functions involved in cell development, growth, proliferation, and cell morphology. In addition, we found that the MFG transcriptome was comparable to the metabolic gene expression profile described for the lactating mouse mammary gland. This paper is the first to describe the MFG transcriptome in sequential human samples over a 24 h period, providing valuable insights into gene expression in the human MEC.

Mechanisms to conserve glucose in lactating women during a 42-h fast.

Little is known about how lactating women accommodate for their increased glucose demands during fasting to avoid maternal hypoglycemia. The objective of this study was to determine whether lactating women conserve plasma glucose by reducing maternal glucose utilization by increasing utilization of FFA and ketone bodies and/or increasing gluconeogenesis and mammary gland hexoneogenesis. Six healthy exclusively breastfeeding women and six nonlactating controls were studied during 42 h of fasting and 6 h of refeeding. Glucose and protein kinetic parameters were measured using stable isotopes and GCMS and energy expenditure and substrate oxidation using indirect calorimetry. After 42 h of fasting, milk production decreased by 16% but remained within normal range. Glucose, insulin, and C-peptide concentrations decreased with the duration of fasting in both groups but were lower (P < 0.05) in lactating women. Glucagon, FFA, and beta-hydroxybutyrate concentrations increased with fasting time (P < 0.001) and were higher (P < 0.0001) in lactating women during both fasting and refeeding. During 42 h of fasting, gluconeogenesis was higher in lactating women compared with nonlactating controls (7.7 +/- 0.4 vs. 6.5 +/- 0.2 micromol kg(-1) min(-1), P < 0.05), whereas glycogenolysis was suppressed to similar values (0.4 +/- 0.1 vs. 0.9 +/- 0.2 micromol kg(-1) min(-1), respectively). Mammary hexoneogenesis did not increase with the duration of fasting. Carbohydrate oxidation was lower and fat and protein oxidations higher (P < 0.05) in lactating women. In summary, lactating women are at risk for hypoglycemia if fasting is extended beyond 30 h. The extra glucose demands of extended fasting during lactation appear to be compensated by increasing gluconeogenesis associated with ketosis, decreasing carbohydrate oxidation, and increasing protein and FFA oxidations.

Short-term effects of recombinant human growth hormone and feeding on gluconeogenesis in humans.

After a short-term fast, lactating women have increased rates of glucose production but not gluconeogenesis (GNG) despite relative hypoinsulinemia. We explored the effects of non-insulin-dependent increase in glucose utilization and recombinant human growth hormone (rhGH) on glucose production, glycogenolysis, and GNG in both the fed and overnight-fasted condition. Six controls and 7 lactating women were studied twice, in random order, after 7 days of saline or rhGH. Glucose kinetics and GNG were measured using [U-(13)C]glucose mass isotopomer distribution analysis. The rhGH increased milk production in the lactating women and insulin-like growth factor (IGF) in both groups. Glycogenolysis and GNG were higher in fasting lactating women than controls after either saline or rhGH (P < .05). After rhGH administration, GNG remained higher (P < .02) in the lactating women than controls. Gluconeogenesis was not suppressed in either group during 5 hours of continuous meal ingestion, despite a 5-fold increase in plasma insulin. Lactating women had similar glucose but lower insulin and C-peptide concentrations than controls after both rhGH and saline treatment (P < .01), although rhGH decreased (P < .01) insulin sensitivity in both groups (P < .05). Gluconeogenesis is not affected by short-term increases in insulin and/or rhGH, which suggests a fundamental rethinking of the role of insulin in acutely regulating GNG.

Measurement of gluconeogenesis using glucose fragments and mass spectrometry after ingestion of deuterium oxide.

We report a new method to measure the fraction of glucose derived from gluconeogenesis using gas chromatography-mass spectrometry and positive chemical ionization. After ingestion of deuterium oxide by subjects, glucose derived from gluconeogenesis is labeled with deuterium. Our calculations of gluconeogenesis are based on measurements of the average enrichment of deuterium on carbon 1, 3, 4, 5, and 6 of glucose and the deuterium enrichment in body water. In a sample from an adult volunteer after ingestion of deuterium oxide, fractional gluconeogenesis using the "average deuterium enrichment method" was 48.3 +/- 0.5% (mean +/- SD) and that with the C-5 hexamethylenetetramine (HMT) method by Landau et al. (Landau BR, Wahren J, Chandramouli V, Schumann WC, Ekberg K, Kalhan SC; J Clin Invest 98: 378-385, 1996) was 46.9 +/- 5.4%. The coefficient of variation of 10 replicate analyses using the new method was 1.0% compared with 11.5% for the C-5 HMT method. In samples derived from an infant receiving total parenteral nutrition, fractional gluconeogenesis was 13.3 +/- 0.3% using the new method and 13.7 +/- 0.8% using the C-5 HMT method. Fractional gluconeogenesis measured in six adult volunteers after 66 h of continuous fasting was 83.7 +/- 2.3% using the new method and 84.2 +/- 5.0% using the C-5 HMT method. In conclusion, the average deuterium enrichment method is simple, highly reproducible, and cost effective. Furthermore, it requires only small blood sample volumes. With the use of an additional tracer, glucose rate of appearance can also be measured during the same analysis. Thus the new method makes measurements of gluconeogenesis available and affordable to large numbers of investigators under conditions of low and high fractional gluconeogenesis ( approximately 10 to approximately 90) in all subject populations.

Short-term high dietary fructose intake had no effects on insulin sensitivity and secretion or glucose and lipid metabolism in healthy, obese adolescents.

UNLABELLED: There is virtually no information on the metabolic impact of dietary fructose intake in adolescents despite their high fructose consumption, particularly via sweetened beverages. AIM: To determine the short-term metabolic effects of dietary fructose intake in obese adolescents. METHODS: Six volunteers (3 M/3 F; 15.2 +/- 0.5 yr; 35 +/- 2 kg/m2; 39 +/- 2% body fat) were studied twice following 7 d of isocaloric, isonitrogenous high carbohydrate (60% CHO; 25% fat) diets with fructose accounting for 6% and 24% of total energy intake, respectively (random order). Insulin sensitivity and secretion were analyzed by the stable labeled intravenous glucose tolerance test and glucose and lipid kinetics using GCMS. RESULTS: A fourfold increase in dietary fructose intake did not affect insulin sensitivity or secretion, glucose kinetics, lipolysis or glucose, insulin, C-peptide, triglycerides, HDL- and LDL-cholesterol concentrations. CONCLUSIONS: In the short term, when energy intake is constant, dietary fructose per se is not a contributor to insulin resistance and hypersecretion in obese adolescents.

Glucose fluxes during OGTT in adolescents assessed by a stable isotope triple tracer method.

UNLABELLED: Virtually no information is available on glucose fluxes during a meal or glucose ingestion in adolescents. AIM: To use a triple tracer approach to measure rates of appearance of ingested glucose (Ra(ogtt)), endogenous glucose production (EGP) and glucose disappearance (Rd) following an oral glucose bolus in adolescents. METHODS: Eleven adolescents (4 M/7 F, 15 +/- 1 yr; 67.3 +/- 4.7 kg; 24 +/-2 kg/m2) underwent a frequent sampled oral glucose tolerance test (OGTT) (labelled with [6,6-2H2]glucose) combined with intravenous infusion of [1-(13)C]glucose and [U-(13)C6]glucose following an overnight fast. Formulas were developed to estimate glucose fluxes using one- or two-compartment models. RESULTS: During the 7 h following the OGTT bolus, 9.8 +/- 2.3% of the ingested glucose was extracted by the liver, EGP was suppressed by 45 +/- 4% and Rd increased by 21 +/- 5%. CONCLUSIONS: The triple tracer method provided accurate assessment of Ra(ogtt), EGP and Rd fluxes during an OGTT in adolescents. Thus, this method might provide novel insight on postprandial glucose fluxes in children/adolescents under various conditions.

Obesity-Related Metabolic Risk in Sedentary Hispanic Adolescent Girls with Normal BMI.

Hispanic adolescent girls with normal BMI frequently have high body fat %. Without knowledge of body fat content and distribution, their risk for metabolic complications is unknown. We measured metabolic risk indicators and abdominal fat distribution in post-pubertal Hispanic adolescent girls with Normal BMI (N-BMI: BMI < 85th percentile) and compared these indicators between girls with Normal BMI and High Fat content (N-BMI-HF: body fat ≥ 27%; n = 15) and Normal BMI and Normal Fat content (N-BMI-NF: body fat < 27%; n = 8). Plasma concentrations of glucose, insulin, adiponectin, leptin and Hs-CRP were determined. Insulin resistance was calculated using an oral glucose tolerance test. Body fat % was measured by DXA and subcutaneous, visceral and hepatic fat by MRI/MRS. The N-BMI-HF girls had increased abdominal and hepatic fat content and increased insulin resistance, plasma leptin and Hs-CRP concentrations (p < 0.05) as compared to their N-BMI-NF counterparts. In N-BMI girls, insulin resistance, plasma insulin and leptin correlated with BMI and body fat % (p < 0.05). This research confirms the necessity of the development of BMI and body fat % cut-off criteria per sex, age and racial/ethnic group based on metabolic risk factors to optimize the effectiveness of metabolic risk screening procedures.

Effects of dietary macronutrient intake on insulin sensitivity and secretion and glucose and lipid metabolism in healthy, obese adolescents.

CONTEXT: Adolescent obesity is a serious public health concern. OBJECTIVE: The aim of the study was to determine whether obese adolescents can adapt metabolically to changes in dietary macronutrient intake. PATIENTS AND DESIGN: Using a random cross-over design, 13 healthy obese volunteers (six boys and seven girls; age, 14.7 +/- 0.3 yr; body mass index, 34 +/- 1 kg/m2; body fat, 42 +/- 1%) were studied twice after 7 d of isocaloric, isonitrogenous diets with 60% carbohydrate (CHO) and 25% fat (high CHO), or 30% CHO and 55% fat (low CHO). MAIN OUTCOME MEASURES AND METHODS: Glucose metabolism, insulin sensitivity, and first- and second-phase insulin secretory indices were measured by stable isotope techniques and the stable labeled iv glucose tolerance test. The results were compared with those of previously studied lean adolescents. RESULTS: Obese adolescents increased first- and second-phase insulin secretory indices by 18 (P = 0.05) and 36% (P = 0.05), respectively, to maintain normoglycemia during the high-CHO diet because they failed to increase insulin sensitivity as did the lean adolescents. Regardless of diet, in obese adolescents, insulin sensitivity was half (P < 0.05) and first- and second-phase insulin secretory indices twice (P < 0.01), compared with the the corresponding values in lean subjects. In obese adolescents, gluconeogenesis increased by 32% during the low-CHO (high-fat diet) (P < 0.01). CONCLUSION: In obese adolescents, insulin secretory demands were increased regardless of diet. Failure to increase insulin sensitivity while receiving a high-CHO diet required a further increase in insulin secretion, which may lead to earlier beta-cell failure. A low-CHO/high-fat diet resulted in increased gluconeogenesis, which may be a prelude to the increased glucose production and hyperglycemia observed in type 2 diabetics.

Impact of duration of infusion and choice of isotope label on isotope recycling in glucose homeostasis.

The purposes of this study were to quantify the impact of the duration of infusion and choice of stable isotope of glucose on measures of glucose rate of appearance (glucose R(a)) and to determine whether the differences observed were due to tracer recycling via the glycogen pool (direct pathway) or gluconeogenesis (indirect pathway). Six healthy adult volunteers were studied on four occasions in the postabsorptive state during infusions of [1-(13)C]glucose and [6,6-(2)H(2)]glucose: 2.5-h infusion of both (A), and 2.5-h infusion of one (B) and 14.5-h infusion of the other isotope (C), and 5-h infusion of [6,6-(2)H(2)]glucose and 2.5-h infusion of [1-(13)C]glucose (D). Infusion of both isotopes for 2.5 h resulted in similar glucose R(a) values. When compared with a 14.5-h infusion, the 2.5-h glucose tracer infusion overestimated glucose R(a) by 26-35%. Glucose (13)C recycled via the Cori cycle, resulting in slower decay from the plasma pool and longer half-life of [1-(13)C]glucose compared with [6,6-(2)H(2)]glucose. There was no detectable release of [(13)C]glucose or [(2)H(2)]glucose tracer into the plasma pool after administration of glucagon. These data demonstrate that glucose R(a) varies not as a result of isotope cycling but as a result of differences in duration of isotope infusion regardless of the isotope used. This is most likely due to incomplete isotope and substrate equilibration with the 2.5-h infusion. The potential error was reduced by nearly 80% using a 5-h infusion of [6,6-(2)H(2)]glucose. These studies demonstrate that the duration of isotope infusion has significantly greater impact on quantitation of glucose R(a) than does the selection of isotope.

Measurements of Gluconeogenesis and Glycogenolysis: A Methodological Review.

Gluconeogenesis is a complex metabolic process that involves multiple enzymatic steps regulated by myriad factors, including substrate concentrations, the redox state, activation and inhibition of specific enzyme steps, and hormonal modulation. At present, the most widely accepted technique to determine gluconeogenesis is by measuring the incorporation of deuterium from the body water pool into newly formed glucose. However, several techniques using radioactive and stable-labeled isotopes have been used to quantitate the contribution and regulation of gluconeogenesis in humans. Each method has its advantages, methodological assumptions, and set of propagated errors. In this review, we examine the strengths and weaknesses of the most commonly used stable isotopes methods to measure gluconeogenesis in vivo. We discuss the advantages and limitations of each method and summarize the applicability of these measurements in understanding normal and pathophysiological conditions.

Hexoneogenesis in the human breast during lactation.

Lactose is the major osmotic agent in milk. Therefore, lactose synthesis indirectly regulates milk volume. The aim of this study was to determine the source of glucose and galactose in lactose. Six healthy lactating women were studied twice, during a 24-h fast and during ingestion of a mixed macronutrient drink (Sustacal) using [U-13C]glucose and [2-13C]glycerol. Six additional lactating women were studied on one single occasion during ingestion of glucose labeled with [1-13C]glucose. Using the ratios of [13C6] enrichments of glucose in lactose and plasma glucose and that of galactose in lactose and plasma glucose, we determined that 98 +/- 3% of glucose and 68 +/- 7% of galactose in lactose were derived from plasma glucose in the fed state, and 72 +/- 4 and 51 +/- 3%, respectively, after a 24-h fast. Virtually identical results (97 +/- 6 and 64 +/- 4%, respectively) were obtained during the glucose feeding study. On the basis of the [13C1] enrichment of glucose and galactose in lactose (derived from [2-13C]glycerol), glycerol contributes to the production of galactose but not glucose within the breast. Thus, plasma glucose is an important source of lactose, but significant amounts of glucose and galactose in lactose are generated within the breast, a process denoted hexoneogenesis. In this process, glycerol is a precursor for milk galactose but not glucose.

Effects of dietary macronutrient content on glucose metabolism in children.

Effects of carbohydrate, fat, and fructose intake on substrate and hormone concentrations, glucose production, gluconeogenesis, and insulin sensitivity were determined in healthy, nonobese prepubertal children (n = 12) and adolescents (n = 24) using a cross-over design. In one group (12 prepubertal children and 12 adolescents), subjects were studied after 7 d of isocaloric, isonitrogenous diets providing either 60% carbohydrate and 25% fat [high carbohydrate (H(CHO))/low fat (L(F))] or 30% carbohydrate and 55% fat [low carbohydrate (L(CHO))/high fat (H(F))], and in a second group (12 adolescents) H(CHO)/L(F) diets containing either 40% or 10% fructose was used. All subjects adapted to changes in carbohydrate and fat intakes primarily by appropriately adjusting their substrate oxidation rates to match the intakes, with only minor changes in parameters of glucose metabolism. Changing from a L(CHO)/H(F) to H(CHO)/L(F) diet resulted in increased insulin sensitivity (stable labeled iv glucose tolerance test) in adolescents [from 3.2 +/- 0.7 x 10(-4) to 5.0 +/- 1.4 x 10(-4) (min(-1))/( micro U.ml(-1)) (mean +/- SE)] but not in prepubertal children [9.4 +/- 2.5 x 10(-4) to 9.9 +/- 1.5 x 10(-4) (min(-1))/( micro U.ml(-1))], whereas beta-cell sensitivity was unaffected in both groups. Insulin sensitivity was higher in prepubertal children than in adolescents (P < 0.05). The dietary fructose content did not affect any measured parameter. We conclude that in the short term, dramatic changes in fat and carbohydrate intakes (regardless of fructose content) did not adversely affect glucose and lipid metabolism in healthy nonobese children. In the adolescents, the high carbohydrate diet resulted in increased insulin sensitivity, thus facilitating insulin-mediated glucose uptake.

Metabolic adaptation to high-fat and high-carbohydrate diets in children and adolescents.

BACKGROUND: Difficulty adapting to high-fat (HF) and high carbohydrate (HC) diets may predispose children to obesity and diabetes. OBJECTIVE: We tested the hypothesis that children have metabolic flexibility to adapt to HF and HC diets. DESIGN: In protocol 1, 12 children aged 6-9 y and 12 adolescents aged 13-16 y were randomly assigned in a crossover design to consume low-fat (LF), HC (25% and 60% of energy, respectively) or HF, low-carbohydrate (LC) (55% and 30% of energy, respectively) diets. In protocol 2, 12 adolescents aged 13-16 y were randomly assigned in a crossover design to consume an LF-HC diet with 11% or 40% of carbohydrate as fructose. Total energy expenditure, nonprotein respiratory quotients (NPRQs), and substrate utilization were measured by using 24-h calorimetry. Effects of sex, puberty, body fat (dual-energy X-ray absorptiometry), intraabdominal fat (magnetic resonance imaging), and fitness on substrate utilization were tested. RESULTS: Substrate utilization was not affected by puberty, body fat, intraabdominal fat, or fitness. Total energy expenditure was not affected by diet. In protocol 1, NPRQs and carbohydrate and fat utilization were significantly affected by diet (P = 0.001) and sex (P = 0.005). NPRQs and carbohydrate utilization increased with the LF-HC diet. NPRQs decreased and fat utilization increased with the HF-LC diet; changes in substrate utilization were less pronounced in females than in males. In protocol 2, 24-h NPRQs and 24-h substrate utilization were not significantly affected by fructose, although net carbohydrate and fat utilization were significantly lower and higher, respectively, with the high-fructose diet during fasting (P = 0.01) and in the subsequent feeding period (P = 0.05). CONCLUSION: Healthy, nonobese children and adolescents adapt appropriately to HF and HC diets.

Splanchnic galactose extraction is regulated by coingestion of glucose in humans.

When compared with galactose alone, coingestion of glucose with galactose decreases plasma galactose. The objective of this study was to determine if this was due to increased peripheral clearance or increased first pass clearance of galactose. Five adult volunteers were studied on 2 occasions during infusion of [6,6-(2)H(2)]glucose and [1-(13)C]galactose and ingestion of galactose alone at 11, 22, and 33 micromol x kg(-1) x min(-1) or galactose plus glucose at 11, 22, and 33 micromol x kg(-1) x min(-1) of each sugar. At 33 micromol x kg(-1) x min(-1) of galactose alone (1) plasma galactose increased to 2.3 +/- 0.3 mmol/L and galactose rates of appearance (Ra) to 18.3 +/- 1.6 micromol x kg(-1) x min(-1); (2) plasma glucose and glucose Ra were unaffected; (3) splanchnic extraction of galactose plateaued at approximately 15 micromol x kg(-1) x min(-1); and (4) galactose became the primary source of glucose Ra (75% +/- 9%). Coingestion of glucose and galactose at 33 micromol x kg(-1) x min(-1) each resulted in (1) decreased plasma galactose (0.3 +/- 0.1 mmol/L) and galactose Ra (6.4 +/- 1.8 micromol x kg(-1) x min(-1)); (2) increased plasma glucose and insulin; (3) doubling of splanchnic extraction of galactose; and (4) decreased contribution of galactose to glucose Ra (11% +/- 4%). We conclude that coingestion of glucose with galactose increases the splanchnic extraction, but decreases the conversion of galactose to glucose.

Galactose promotes fat mobilization in obese lactating and nonlactating women.

BACKGROUND: Galactose consumption as the only carbohydrate source results in little increase in plasma glucose and insulin concentrations when compared with fasting. Lower insulin might promote endogenous lipolysis during meal absorption, which may facilitate fat loss. OBJECTIVE: The objective was to test the hypothesis that consumption of an isocaloric, isonitrogenous galactose drink will result in higher rates of lipolysis and fat oxidation than consumption of a glucose drink in obese lactating and nonlactating women. DESIGN: Seven healthy, obese, exclusively breastfeeding women and 7 healthy, obese, nonlactating women were studied on 2 occasions according to a randomized, crossover, single-blinded design. Subjects received drinks providing ≈70% of the daily estimated energy requirement, of which 60% was either glucose or galactose. The primary outcomes were the rate of appearance (Ra) of glycerol and palmitate, and the secondary outcomes were glucose Ra, milk production, energy expenditure, and substrate oxidation. RESULTS: Plasma glucose and insulin concentrations were lower (P < 0.05) and those of glycerol, palmitate, free fatty acids, and triglycerides were higher (P < 0.05) during galactose than during glucose feeding in both nonlactating and lactating women. During galactose feeding, glucose Ra was lower (P < 0.01) and glycerol, palmitate, and free fatty acid Ra were higher (P < 0.01) in both groups. During galactose feeding in all women combined, fat oxidation was higher (P = 0.01) and protein oxidation was lower (P < 0.01). Milk production, energy expenditure, and carbohydrate oxidation were similar between glucose and galactose feeding. CONCLUSIONS: Galactose consumption is associated with higher endogenous fat mobilization and oxidation during meal absorption. Long-term studies are required to determine whether galactose as an exclusive carbohydrate source would promote body fat loss in obese subjects.

Gluconeogenesis continues in premature infants receiving total parenteral nutrition.

OBJECTIVE: To determine the contribution of total gluconeogenesis to glucose production in preterm infants receiving total parenteral nutrition (TPN) providing glucose exceeding normal infant glucose turnover rates. STUDY DESIGN: Eight infants (0.955±0.066 kg, 26.5±0.5 weeks, 4±1 days) were studied while receiving routine TPN. The glucose appearance rate (the sum of rates of glucose infusion and residual glucose production) and gluconeogenesis were measured by stable isotope-gas chromatography-mass spectrometry techniques using deuterated water and applying the Chacko and Landau methods. RESULTS: Blood glucose ranged from 5.2 to 14.3 mmol/l (94-257 mg/dl) and the glucose infusion rate from 7.4 to 11.4 mg/kg per min, thus exceeding the normal glucose production rates (GPR) of newborn infants in most of the babies. The glucose appearance rate was 12.4±0.6 and GPR 2.1±0.3 mg/kg per min. Gluconeogenesis as a fraction of the glucose appearance rate was 11.2±1.1% (Chacko) and 10.5±1.2% (Landau) (NS) and the rate of gluconeogenesis was 1.35±0.15 mg/kg per min (Chacko) and 1.29±0.14 mg/kg per min (Landau) (NS). Gluconeogenesis accounted for 73±11% and 68±10 (NS) of the GPR for the two methods, respectively. Gluconeogenesis and glycogenolysis were not affected by the total glucose infusion rate, glucose concentration, gestational age or birth weight. Glucose concentration correlated with the total glucose infusion rate and gestational age (combined R(2)=0.79, p=0.02). CONCLUSIONS: Gluconeogenesis is sustained in preterm infants receiving routine TPN providing glucose at rates exceeding normal infant glucose turnover rates and accounts for the major part of residual glucose production. Gluconeogenesis is not affected by the glucose infusion rate or blood glucose concentration.