bluuebunny/crossref_metadata_2025 huggingface dataset:
Insulin and hyperaminoacidemia regulate by a different mechanism leucine turnover and oxidation in obesity
Authors: Luzi L. , Castellino P. , DeFronzo R. A.
Published: December 2017

Seven normal glucose-tolerant obese subjects [ideal body weight (IBW) = 161%] and 18 controls (IBW = 102%) were studied with the euglycemic insulin clamp (10 and 40 mU.m-2.min-1) technique, [14C]leucine infusion, and indirect calorimetry to examine if the insulin resistance with respect to glucose metabolism extends to amino acid/protein metabolism. In the basal state, total plasma amino acid and leucine concentrations, endogenous leucine flux (ELF), leucine oxidation (LO), and nonoxidative leucine disposal (NOLD) were similar in obese and control subjects. During both low (10 mU.m-2.min-1)- and higher (40 mU.m-2.min-1)-dose insulin clamp studies, insulin-mediated glucose uptake was reduced in obese vs. control subjects (P < 0.01). During the last hour of the higher-dose insulin clamp step, the decrease in total plasma amino acids, branched-chain amino acids, and leucine concentration was impaired in obese vs. control subjects (P < 0.01). However, suppression of ELF and NOLD was similar in both groups. During the low-dose insulin clamp, the decrease in plasma leucine concentration, LO, and ELF all were impaired (P < 0.01). A second study was performed in which the total plasma amino acid concentration was increased two- to threefold in both groups. Under these conditions of low plasma insulin/high amino acid levels, LO and NOLD increased similarly in obese and control subjects. In conclusion, insulin resistance is a common feature of both glucose and protein metabolism in obesity. The defect in protein metabolism is characterized by an impairment of the ability of insulin to inhibit proteolysis; the stimulatory effect of hyperaminoacidemia on protein synthesis is intact in obesity.
Androgens contribute to the stimulation of cancellous bone formation by ovarian hormones in female rats
Authors: Gallagher A. C. , Chambers T. J. , Tobias J. H.
Published: December 2017

We investigated whether androgens, for which the ovaries are the major source in female rats, contribute to the stimulation of cancellous bone formation by ovarian hormones in female rats. Ovariectomized animals were administered 5alpha-dihydrotestosterone (DHT; 10 and 100 microgram/kg) by daily subcutaneous injection for 13 days, after which histomorphometric analysis was performed at the proximal tibial metaphysis. To prevent ovariectomy from stimulating bone turnover, and hence complicating the interpretation of changes in bone formation, animals were also given the resorption inhibitor 3-amino-1-hydroxypropylidene-1-bisphosphonate. We found that ovariectomy markedly suppressed cancellous bone formation, which was partially prevented by DHT (100 microgram/kg). To further address whether androgens contribute to the stimulation of bone formation by ovarian hormones, we treated intact and ovariectomized female rats with the androgen antagonist flutamide (15 mg/kg/day) for 28 days. Whereas flutamide had no effect in ovariectomized rats, it significantly reduced cancellous bone formation in intact animals. We conclude that, in the female rat, androgens contribute under physiological conditions to the stimulation of cancellous bone formation by ovarian hormones.
IL-1 receptor antagonist attenuates sepsis-induced alterations in the IGF system and protein synthesis
Authors: Lang C. H. , Fan J. , Cooney R. , Vary T. C.
Published: December 2017

The purpose of the present investigation was to determine whether endogenously produced interleukin (IL)-1 mediates the changes in insulin-like growth factor (IGF) I and IGF binding proteins (IGFBP) induced by chronic abdominal sepsis in rats and to correlate the changes in the IGF system with the alternations in protein synthesis. A constant infusion of IL-1 receptor antagonist (IL-1ra) was begun after the induction of sepsis and was continued for 5 days. Sepsis decreased IGF-I levels in the blood, liver, and gastrocnemius muscle, increased the content in the kidney, and did not alter IGF-I levels in heart, jejunum, and spleen. IL-1ra attenuated the sepsis-induced decrease in plasma IGF-I and completely prevented the changes in IGF-I observed in liver, kidney, and the gastrocnemius. IGFBP-1 was increased in the blood, liver, and muscle of septic rats. IL-1ra prevented this increase in IGFBP-1 in blood and liver but not in muscle. The rate of in vivo protein synthesis was decreased in the gastrocnemius and kidney and unaltered in the heart, liver, jejunum, and spleen. A strong linear correlation existed between levels of IGF-I and the rate of protein synthesis determined simultaneously in the gastrocnemius. These results provide evidence for the role of IL-1 as an endogenous mediator of the sepsis-induced changes in IGF-I and IGFBP-1 and suggest that the accompanying changes in muscle protein synthesis are partially mediated via changes in IGF-I.
Elevated plasma glucose 2 h postchallenge predicts defects in beta-cell function
Authors: Byrne M. M. , Sturis J. , Sobel R. J. , Polonsky K. S.
Published: December 2017

Studies were performed in subjects with no known family history of diabetes, normoglycemic subjects who have first-degree relatives with non-insulin-dependent diabetes mellitus (NIDDM), and subjects with nondiagnostic oral glucose tolerance tests (NDX) or impaired glucose tolerance (IGT). Insulin sensitivity index (SI) was similar in all four groups. However, a number of defects in insulin secretion were seen in the NDX and IGT groups, including reduced first-phase insulin secretory responses in intravenous glucose in relation to the degree of insulin resistance, and reduced normalized spectral power of insulin secretion during oscillatory glucose infusion. The latter finding demonstrates a decreased ability of the beta-cell to detect and respond to the successive increases and decreases in glucose and therefore to be entrained by the exogenous glucose infusion. The ability of a low-dose glucose infusion to prime the insulin secretory response to a subsequent glucose stimulus was normal in subjects with IGT but reduced or absent in subjects with overt NIDDM. These studies demonstrate that a number of alterations in beta-cell function are detectable in nondiabetic first-degree relatives of subjects with NIDDM with mild elevations in the 2-h postchallenge glucose level, and these abnormalities antedate the onset of overt hyperglycemia and clinical diabetes.
A new procedure for the isolation of plasma membranes, T tubules, and internal membranes from skeletal muscle
Authors: Dombrowski L. , Roy D. , Marcotte B. , Marette A.
Published: December 2017

A new subcellular fractionation procedure for the simultaneous isolation of plasma membranes and transverse (T) tubule membranes from a rat skeletal muscle was developed. This new technique allows the isolation and separation of plasma membranes and T tubules in distinct subcellular fractions, as revealed by the membrane distribution of enzymatic and immunologic markers of both cell surface compartments. The procedure also yields a novel membrane fraction that is devoid of markers of both surface domains but is markedly enriched with GLUT-4 glucose transporters, thus strongly suggesting that it represents an intracellular pool of GLUT-4. Using this new procedure, we found that acute in vivo insulin administration (30 min) increased GLUT-4 protein content in the plasma membrane and a T tubule fraction (by approximately 80%), whereas a smaller elevation (35%) was observed in another fraction enriched with T tubules. Insulin induced a concomitant reduction (approximately 40%) in GLUT-4 abundance in the intracellular fraction. These results further support the hypothesis that T tubules are involved in the regulation of glucose transport in skeletal muscle. This novel fractionation method will be useful in investigating the regulation of muscle GLUT-4 transporters in other physiological and disease states such as diabetes, where defective translocation of the transporter protein to either one or both cell surface domains is suspected to occur.
Osteocytic expression of mRNA for c-fos and IGF-I: an immediate early gene response to an osteogenic stimulus
Authors: Lean J. M. , Mackay A. G. , Chow J. W. , Chambers T. J.
Published: December 2017

We analyzed the expression, during the osteogenic response of bone to mechanical stimulation, of insulin-like growth factor I (IGF-I), a growth factor implicated in bone formation, and c-fos, a protooncogene in which disordered regulation specifically affects bone. Both genes were strongly expressed in osteocytes of mechanically stimulated but not control bones within 30 min of the osteogenic stimulus. IGF-I mRNA expression increased up to 6 h, was restricted to osteocytes, and was strongly suppressed by indomethacin. Although early IGF-I mRNA expression was resistant to cycloheximide, there was a degree of suppression after 6 h, raising the possibility that IGF-I expression might be prolonged by autocrine mechanisms. c-fos mRNA was increased both in osteocytes and on bone surfaces. At both sites, c-fos expression was transient, prolonged by cycloheximide, and was strongly stimulated even in the presence of indomethacin. Thus osteocytes respond to mechanical stimulation with immediate prolonged expression of IGF-I and immediate transient expression of c-fos, implicating osteocytes in the osteogenic response to mechanical stimulation. Moreover, the different spatial distribution and indomethacin sensitivity of c-fos and IGF-I gene expression suggest that at least two signaling pathways are activated in osteocytes during this process.
Somatostatin infusion suppresses GH secretory burst frequency and mass in normal men
Authors: Calabresi E. , Ishikawa E. , Bartolini L. , Delitala G. , Fanciulli G. , Oliva O. , Veldhuis J. D. , Serio M.
Published: December 2017

In attempting to elucidate the neuroendocrine mechanisms that regulate pulsatile growth hormone (GH) secretion, we measured serum GH concentrations by an ultrasensitive immunofluorometric method in blood collected every 10 min for 8 h in 11 young healthy male volunteers (age range 21-31 yr) before and during somatostatin (SS) administration (an iv bolus dose of 350 micrograms followed by a continuous infusion at the rate of 6 micrograms.kg-1.h-1, which increases the circulating SS levels to approximately 570 pg/ml). Pulsatile GH secretion was analyzed using the computer-assisted pulse detection program cluster method and deconvolution analysis. The area and frequency of GH peaks were significantly reduced during SS infusion compared with basal values, but detectable pulsatile episodes were still present. These data suggest that, in adult males, SS controls pulsatile GH secretion and can decrease the mass and frequency of GH secretory bursts.
Direct effects of catecholamines on hepatic glucose production in conscious dog are due to glycogenolysis
Authors: Chu C. A. , Sindelar D. K. , Neal D. W. , Cherrington A. D.
Published: December 2017

The effects of catecholamines (CATS) infused into the hepatic portal vein were studied in ten 18-h-fasted conscious dogs. Glucose production (GP) and gluconeogenesis (GNG) were assessed using tracer ([3H]glucose, [14C]alanine) and arteriovenous difference techniques. Each experiment consisted of a 100-min equilibration, a 40-min basal, and two 90-min test periods. A pancreatic clamp (somatostatin + basal portal insulin and glucagon) was used to fix insulin and glucagon at basal levels. Propranolol (1 microgram.kg-1.min-1) and phentolamine (2 micrograms.kg-1.min-1) were infused intraportally during both test periods of the blockade group while a carrier solution was infused in the control group. Norepinephrine (NE; 100 ng.kg-1.min-1) and epinephrine (Epi; 40 ng.kg-1.min-1) were infused intraportally during the second test period of both protocols. Portal NE (70 +/- 46 to 8,404 +/- 674 and 162 +/- 57 to 6,530 +/- 624 pg/ml, respectively) and portal Epi (21 +/- 11 to 3,587 +/- 309 and 29 +/- 6 to 2,989 +/- 406 pg/ml, respectively) rose in the control and adrenergic blockade groups, respectively. The increases in arterial NE and Epi were modest in both groups. Intraportal infusion of CATS increased GP from 2.1 +/- 0.2 to 6.2 +/- 1.0 mg.kg-1.min-1 in the control group but did not change it (2.7 +/- 0.4 to 2.7 +/- 0.3 mg.kg-1.min-1) in the blockade group. Portal CATS had no effect on GNG in the presence or absence of adrenergic blockade (GNG rose from 0.7 +/- 0.2 to 0.9 +/- 0.2 and 0.8 +/- 0.2 to 1.0 +/- 0.2 mg.kg-1.min-1 in the control and blockade groups, respectively). In conclusion, portal infusion of catecholamines significantly augmented GP by selectively stimulating glycogenolysis. The increase in hepatic GP could be completely inhibited by intraportal adrenergic blockade.
Effects of circulating IGF-I on glucose and amino acid kinetics in the ovine fetus
Authors: Liechty E. A. , Boyle D. W. , Moorehead H. , Lee W. H. , Bowsher R. R. , Denne S. C.
Published: December 2017

To investigate the role of insulin-like growth factor I (IGF-I) in the regulation of fetal metabolism, the kinetics of leucine, phenylalanine, and glucose were assessed in the chronically catheterized ovine fetus (0.85 gestation) before and during infusion of recombinant human IGF-I (rhIGF-I). Substrate kinetics were determined by tracer dilution. rhIGF-I was infused at 6.7 nmol.kg fetus-1.h-1. Fetal insulin and growth hormone concentrations were significantly decreased by 50% during rhIGF-I infusion. Net umbilical glucose uptake was unchanged, and glucose rate of appearance increased in the fed state only. There were no changes in the net umbilical uptakes of leucine or phenylalanine, but the rates of appearance of both declined during rhIGF-I infusion, indicative of decreased fetal protein breakdown (Ra,Leu 45.4 +/- 1.40 to 40 +/- 1.4 mumol/min in the fed state, 43 +/- 1.5 to 37 +/- 1.5 mumol/min in the fasted state; Ra,Phe 10.7 +/- 0.3 to 10.4 +/- 0.3 mumol/min in the fed state and from 10.7 +/- 0.3 to 9.8 +/- 0.3 mumol/min in the fasted state). Leucine oxidation was also decreased (8.90 +/- 0.76 to 6.52 +/- 0.81 mumol/min, P = 0.025), more so in the fasted than the fed state. These results indicate a significant antiproteolytic endocrine effect for IGF-I in the late-gestation mammalian fetus.
Desensitization of beta-adrenergic receptors in adipocytes causes increased insulin sensitivity of glucose transport
Authors: Green A. , Carroll R. M. , Dobias S. B.
Published: December 2017

To determine the effect of desensitization of adipocyte beta-adrenergic receptors on insulin sensitivity, rats were continuously infused with isoproterenol (50 or 100 micrograms.kg-1.h-1) for 3 days by osmotic minipumps. Epididymal adipocytes were isolated. The cells from treated animals were desensitized to isoproterenol, as determined by response of lipolysis (glycerol release). Binding of [125I]iodocyanopindolol was decreased by approximately 80% in adipocyte plasma membranes isolated from treated rats, indicating that beta-adrenergic receptors were downregulated. Cellular concentrations of Gn alpha and Gi alpha were not altered. Insulin sensitivity was determined by measuring the effect of insulin on glucose transport (2-deoxy-[3H]glucose uptake). Cells from the isoproterenol-infused rats were markedly more sensitive to insulin than those from control rats. This was evidenced by an approximately 50% increase in maximal glucose transport rate in cells from the high-dose isoproterenol-treated rats and by an approximately 40% decrease in the half-maximal effective concentration of insulin in both groups. 125I-labeled insulin binding to adipocytes was not altered by the isoproterenol infusions, indicating that desensitization of beta-adrenergic receptors results in tighter coupling between insulin receptors and stimulation of glucose transport.
Differential permeability of the BBB in acute EAE: enhanced transport of TNT-alpha
Authors: Pan W. , Banks W. A. , Kennedy M. K. , Gutierrez E. G. , Kastin A. J.
Published: December 2017

Impairment of the blood-brain barrier (BBB) in experimental autoimmune encephalomyelitis (EAE) has been frequently attributed to disruption, without much consideration of saturable transport processes. In mice with EAE, we studied the permeability of the BBB to radioactively labeled albumin and sucrose, markers of BBB disruption, and tumor necrosis factor-alpha (TNF-alpha), a cytokine transported across the BBB by a saturable system and thought to play a role in the pathogenesis of EAE. Permeation of the BBB was increased to all three substances during the acutely ill stage, was greatest in the lumbar spine, and returned to normal with recovery. The change in BBB permeability to sucrose was greater than to the larger albumin and is consistent with a partial disruption of the BBB. The enhanced permeability to TNF-alpha was comparable to that for sucrose, even though TNF-alpha is similar in size to albumin. This paradoxically high uptake of TNF-alpha could be explained by an enhancement of its endogenous saturable transport system. Thus the changes in BBB function during EAE extend beyond disruption to include changes in the saturable transport systems for substances involved in the disease process.
Glucose-induced oscillatory insulin secretion in perifused rat pancreatic islets and clonal beta-cells (HIT)
Authors: Cunningham B. A. , Deeney J. T. , Bliss C. R. , Corkey B. E. , Tornheim K.
Published: December 2017

Normal insulin secretion is oscillatory in vivo and from groups of perifused islets. Stimulation of rat islets with different glucose concentrations gave insulin oscillations of similar period (5-8 min) but increasing amplitude. It has been assumed that oscillatory secretion is due to oscillations in intracellular free Ca2+, as seen in single islets and single pancreatic beta-cells. However, when islets were perifused with diazoxide and high KCl to maintain high intracellular free Ca2+, insulin oscillations of similar amplitude and period still occurred on glucose stimulation, although superimposed on elevated basal secretion. Several likely possibilities for a diffusible synchronizing factor were tested, including pyruvate, lactate, ATP, and insulin itself; nevertheless, perifusion with high concentrations of these did not prevent insulin oscillations. Clonal pancreatic beta-cells (HIT) and dissociated islets also exhibited oscillatory insulin secretion, but with the 5- to 8-min period oscillations superimposed on 15- to 20-min period oscillations. These results indicate that the mechanisms for generating and synchronizing insulin oscillations reside in the beta-cell, although the structure of the islet may modulate the oscillation pattern.
Lactate stimulates progesterone secretion via an increase in cAMP production in exercised female rats
Authors: Lu S. S. , Lau C. P. , Tung Y. F. , Huang S. W. , Chen Y. H. , Shih H. C. , Tsai S. C. , Lu C. C. , Wang S. W. , Chen J. J. , Chien E. J. , Chien C. H. , Wang P. S.
Published: December 2017

The effect of exercise on the production of ovarian progesterone was examined in female rats. During in vivo experiments, diestrous rats were catheterized via the right jugular vein (RJV), and blood samples were collected before and after 10, 15, 30, and 60 min of swimming. In addition, blood samples were collected from the RJV before and 2, 5, 10, 15, 30, 60, and 120 min after 10 min of infusion of lactate (13 mg.kg-1.min-1) through the left femoral vein. To explore if lactate modulates progesterone secretion by acting directly on rat ovary or on anterior pituitary gland (AP), an in vitro experiment that mimicked the in vivo condition was performed. The ovarian tissue was challenged with lactate (0.01-10 mM) or porcine follicle-stimulating hormone (1 microgram/ml) and 3-isobutyl-1-methylxanthine (1 mM) for 60 min, and the AP was challenged with lactate ranging from 0.1 to 10 mM or 10 nM gonadotropin-releasing hormone for 30 min. The postexercise levels of plasma glucose, lactate, and progesterone at 10, 15, and 30 min were significantly higher than the corresponding basal levels. Plasma luteinizing hormone (LH) did not change after exercise. An elevation of plasma lactate and progesterone was found at 15 and 30 min subsequent to 10 min of infusion of lactate. Lactate ranging from 0.01 to 10 mM significantly increased ovarian adenosine 3',5'-cyclic monophosphate (cAMP) and progesterone production in a dose-dependent manner. LH concentration in plasma was not changed subsequent to lactate infusion. LH level in media samples was not altered after incubation of AP with lactate. These results suggest that the increase of plasma progesterone level in rats during exercise is independent of LH secretion and at least in part is due directly to a stimulatory effect of lactate on the production of ovarian cAMP.
Expression of IGFBP-2, -3, and -4 mRNA during differentiation of Caco-2 colon epithelial cells
Authors: Hoeflich A. , Yang Y. , Huber S. , Rascher W. , Koepf G. , Blum W. F. , Heinz-Erian P. , Kolb H. J. , Kiess W.
Published: December 2017

Local production of insulin-like growth factor (IGF)-binding proteins (IGFBP) determines the availability of the IGF to the cell and thus regulates IGF action. To find out whether specific patterns of IGFBP gene expression and IGFBP secretion were related to cell growth vs. cell differentiation, expression of IGFBP during long-term culture (21 days, n = 5) of the colon carcinoma cell line Caco-2 was investigated at the mRNA and protein levels. Markers of cell proliferation (increase in DNA, RNA, and protein content) and of differentiation [alkaline phosphatase (AP) activity; creatine kinase (CK) activity] were measured in parallel during long-term culture. IGFBP-2 mRNA expression correlated significantly with markers of proliferation (P < 0.05), whereas IGFBP-3 mRNA expression or IGFBP-3 secretion correlated with markers of differentiation (AP: r = 0.83, P < 0.001; CK: r = 0.45, P < 0.01). Similarly, IGFBP-4 mRNA expression correlated significantly with markers of differentiation (AP: r = 0.34, P < 0.05; CK: r = 0.35, P < 0.05). We hypothesize that IGFBP-3 and -4 are related to differentiation of Caco-2 cells, whereas IGFBP-2 is related to proliferation in Caco-2 cells.
Diurnal rhythms and effects of fasting and refeeding on rat adipose tissue lipoprotein lipase
Authors: Bergo M. , Olivecrona G. , Olivecrona T.
Published: December 2017

The activity of lipoprotein lipase (LPL) in adipose tissue is modulated by changes in the nutritional status. We have measured LPL activity, mass, and mRNA levels in rat adipose tissue during normal feeding cycles, during short- and long-term fasting, and during refeeding after fasting. LPL activity displayed a diurnal rhythm. The activity was highest during the night and early morning, decreased to a minimum during the early afternoon, and then increased again. These changes corresponded to the feeding pattern. The increases and/or decreases resulted from changes in LPL synthetic rate compounded by posttranslational mechanisms. During short-term fasting, LPL specific activity decreased to < 30% of control. The specific activity was restored within 4 h by refeeding. On longer fasting, LPL mRNA decreased. This became significant from 36 h. On refeeding, it took 12 h to restore the mRNA levels, whereas tissue LPL activity and mass could not be fully restored by 36 h of refeeding. These data show that LPL activity during short-term fasting is regulated posttranscriptionally, which allows for quick upregulation after refeeding. On longer fasting, other mechanisms affecting LPL transcription and synthesis come into play, and upregulation after refeeding is slowed down.
Quantitative aspects of interorgan relationships among arginine and citrulline metabolism
Authors: Yu Y. M. , Burke J. F. , Tompkins R. G. , Martin R. , Young V. R.
Published: December 2017

The quantitative roles of the splanchnic region and the kidneys in whole body (WB) arginine and citrulline metabolism were assessed in postabsorptive mongrel dogs with primed constant intravenous infusions of [15N2-guanidino, 5,5-2H2]arginine and [13C-ureido]citrulline or [13C-guanidino]arginine and [15N]urea tracers. Isotope and metabolite concentration balances of arginine and citrulline were measured across the gut, liver, splanchnic region, and kidneys, together with WB arginine and citrulline fluxes and urea production rate. The WB citrulline flux and rate of citrulline to arginine (C-A) conversion were 16 and 9.4 mumol.kg-1.h-1, respectively. Concentration balance of citrulline across kidneys was +8.2 mumol.kg-1.h-1, and metabolism of citrulline by kidneys was 8.7 mumol.kg-1.h-1, which was derived about equally from intestine and liver. The appearance rate of citrulline-derived arginine in renal vein was 6.8 mumol.kg-1.h-1. These three separate estimates of C-A conversion within the kidneys were in good agreement, indicating 40% of blood C-A conversion occurring outside kidneys. These findings of interorgan metabolism are discussed in reference to the current knowledge derived largely from studies in laboratory rodents.
Hepatic VLDL secretion of genetically obese Zucker rats is inhibited by a high-fat diet
Authors: Oussadou L. , Griffaton G. , Kalopissis A. D.
Published: December 2017

Hepatocytes from obese and lean Zucker rats adapted to a control (C) or a high-fat (HF) diet were prepared for the study of fatty acid (FA) uptake, partition between oxidation and esterification, and very low density lipoprotein (VLDL) production. A first 2-h kinetic study showed higher oleate uptake on a C diet by obese rat cells and an almost exclusive esterification to triacylglycerol (TG), VLDL secretion being 2.5-fold higher in obese rat cells and enhanced 1.4-fold in both genotypes in the presence of 0.7 mM oleate vs. 0.1 mM or no oleate. Fat feeding 1) decreased oleate uptake, esterification, incorporation into VLDL-TG, and mass VLDL-TG secretion and 2) abolished the VLDL-TG increase by 0.7 mM oleate. Similar but more pronounced effects were obtained in fat-fed lean animals. A second kinetic study using very short incubation times up to 1 h confirmed that fat feeding decreased oleate uptake and esterification, greatly stimulating its oxidation and production of acetoacetate (obese) or acetoacetate and beta-hydroxybutyrate (lean). Synthesis of lactate and pyruvate greatly decreased under HF feeding, remaining higher in obese rat cells. The drastic inhibition of labeled and total hepatic VLDL-TG secretion in obese and lean Zucker rats by the HF diet could be partly explained by decreased exogenous FA availability for VLDL-TG synthesis through its greater channeling toward oxidation and, indirectly, by the altered hepatocyte metabolic state.
Effects of morning rise in cortisol concentration on regulation of lipolysis in subcutaneous adipose tissue
Authors: Samra J. S. , Clark M. L. , Humphreys S. M. , Macdonald I. A. , Matthews D. R. , Frayn K. N.
Published: December 2017

Cortisol has a well-defined circadian rhythm. The aim of the study was to examine the effect of the morning rise in cortisol concentration on lipolysis in adipose tissue. Ten healthy subjects were studied on two occasions, and six of these were studied on three occasions. During the first two occasions, either a control or cortisol suppression study was performed by using metyrapone, and on the third occasion exogenous cortisol replacement was given in addition to metyrapone. Lipolysis in the subcutaneous adipose tissue of the anterior abdominal wall was studied by measurement of arteriovenous differences. Reduction in the early morning rise in cortisol led to significantly decreased venoarterialized differences for nonesterified fatty acids (P < 0.05) and glycerol (P < 0.01), attributable in part to decreased hormone-sensitive lipase (EC 3.1.1.3) action (P < 0.05) in adipose tissue. At the same time the arterialized plasma triacylglycerol concentration increased (P < 0.005) with a significant reduction in the adipose lipoprotein lipase (EC 3.1.1.34) rate of action (P < 0.05). In the replacement study, values were identical to those of the control study, showing that metyrapone had no nonspecific effects on lipolysis. We conclude that the morning rise in plasma cortisol concentration plays an important role in the regulation of lipolysis in adipose tissue in normal healthy adults.
Effect of magnesium on parathyroid cells: evidence for two sensing receptors or two intracellular pathways?
Authors: Miki H. , Maercklein P. B. , Fitzpatrick L. A.
Published: December 2017

It currently remains controversial as to the intracellular mechanisms coupled to the inhibition of parathyroid hormone (PTH) secretion that are modulated by extracellular divalent cations. To study mechanisms responsible for regulation of PTH release by cations, we investigated the effect of Mg2+ on cytosolic Ca2+ levels ([Ca2+]i) and PTH secretion in single isolated bovine parathyroid cells. Addition of 9.0 mM Mg2+ evoked a spike of [Ca2+]i in approximately 80% of cells in the presence of extracellular Ca2+. Mg2+ decreased steady-state [Ca2+]i, which represented inhibition of influx of extracellular Ca2+ in 13-78% of cells. The percentage of cells that had a decline in steady-state [Ca2+]i after exposure to Mg2+ was dependent on the extracellular Ca2+ concentration. The effect of Mg2+ on intracellular Ca2+ response was dose dependent. Extracellular Mg2+ inhibited PTH secretion in cells that showed decline in steady-state [Ca2+]i, although cells that showed a spike after addition of Mg2+ secreted more PTH than cells that did not show a spike. The spike of [Ca2+]i and decline in steady-state [Ca2+]i that occur in response to extracellular Mg2+ may be caused independently by two distinct mechanisms that differentially regulate secretion of PTH.
Effects and metabolism of fumarate in the perfused rat heart. A 13C mass isotopomer study
Authors: Laplante A. , Vincent G. , Poirier M. , Des Rosiers C.
Published: December 2017

The cardioprotective effects of fumarate have been linked to its metabolism to succinate through both oxidative and reductive pathways. To date, the relative contribution of these pathways is a subject of controversy. To address this question, we designed a protocol with 13C substrates and took advantage of 13C isotopomer analysis by gas chromatography-mass spectrometry. Rat hearts were perfused with 11 mM glucose, 1 mM lactate, 0.2 mM pyruvate, 0.2 mM [1-13C]octanoate, and 0.04 or 0.4 mM [U-13C4]fumarate. On reoxygenation after 40 min of severe hypoxia, hearts perfused with 0.4 mM fumarate showed a better recovery of contractile function and released less lactate dehydrogenase (an index of cellular necrosis) than those perfused with 0.04 mM fumarate. The 13C data showed that, in hypoxic hearts, fumarate conversion to succinate occurred only through reduction, although it accounted for only 16% of total succinate release. Most of the succinate was formed through the oxidation of alpha-ketoglutarate or its precursors (50 +/- 5%) and by another yet-unidentified pathway (34 +/- 4%). These data show that, in a model of hypoxia-reoxygenation, the cardioprotective effects of fumarate were associated with its predominant metabolism to succinate through the reductive pathway.
Urinary biotin analogs increase in humans during chronic supplementation: the analogs are biotin metabolites
Authors: Mock D. M. , Heird G. M.
Published: December 2017

In human subjects, the metabolic origins of bisnorbiotin and biotin sulfoxide were determined by measuring the urinary excretion of each after chronic administration of large oral doses of biotin. For 2 wk, 14 adult volunteers consumed 1,200 micrograms of biotin per day, an amount approximately 20 times the daily dietary intake. With the use of a high-performance liquid chromatography/avidin-binding assay in untimed urine samples obtained before the first dose of biotin and after the 14th dose, concentrations of biotin, bisnorbiotin, and biotin sulfoxide were measured. Excretion was expressed as concentration ratios to urinary creatinine. Bisnorbiotin and biotin sulfoxide excretion increased 85-fold (P < 0.0001) and 114-fold (P < 0.0001), respectively. The molar percentages of bisnorbiotin and biotin sulfoxide decreased from 28 to 14% (P = 0.006) and from 9 to 5% (P = 0.017), respectively. These data provide evidence that the bisnorbiotin and biotin sulfoxide found in human urine are biotin metabolites. Furthermore, we infer that chronic consumption of large amounts of biotin does not substantially saturate the human biotin pathways of biotransformation.
Nutritional status affects branched-chain oxoacid dehydrogenase activity during exercise in humans
Authors: Jackman M. L. , Gibala M. J. , Hultman E. , Graham T. E.
Published: December 2017

We examined the effect of glycogen availability and branched-chain amino acid (BCAA) supplementation on branched-chain oxoacid dehydrogenase (BCOAD) activity during exercise. Six subjects cycled at approximately 75% of their maximal oxygen uptake to exhaustion on three occasions under different preexercise conditions: 1) low muscle glycogen (LOW), 2) low muscle glycogen plus BCAA supplementation (LOW+BCAA), and 3) high muscle glycogen (CON). The LOW trial was performed first, followed by the other two conditions in random order, and biopsies for all trials were obtained at rest, after 15 min of exercise (15 min), and at the point of exhaustion during the LOW trial (49 min). BCOAD activity was not different among the three conditions at rest; however, at 15 min BCOAD activity was higher (P < or = 0.05) for the LOW (31 +/- 5%) and LOW+BCAA (43 +/- 11%) conditions compared with CON (12 +/- 1%). BCOAD activity at 49 min was not different from respective values at 15 min for any condition. These data indicate that BCOAD is rapidly activated during submaximal exercise under conditions associated with low carbohydrate availability. However, there was no relationship between BCOAD activity and glycogen concentration or net glycogenolysis, which suggests that factors other than glycogen availability are important for BCOAD regulation during exercise in humans.
Phenotypic characterization and functional correlation of alpha-MSH binding to pituitary cells
Authors: Zheng T. , Villalobos C. , Nusser K. D. , Gettys T. W. , Faught W. J. , Castano J. P. , Frawley L. S.
Published: December 2017

It is clear that alpha-melanocyte-stimulating hormone (alpha-MSH), released by the hypophysial neurointermediate lobe, is a mediator of suckling-induced prolactin release, but several questions surrounding its role remain unresolved. Accordingly, the objectives of the present study were 1) to establish whether alpha-MSH could bind in a reversible manner to a specific secretory type cell within the adenohypophysis (AP), 2) to resolve the issue of whether the peptide could compete with dopamine for the same receptor binding site, and 3) to seek a functional signaling correlate for alpha-MSH binding. In pursuit of these objectives, we subjected pituitary cells from lactating rats to alpha-MSH receptor autoradiography, AP hormone immunocytochemistry, or digital imaging fluorescence microscopy with fura 2 as a calcium-sensitive probe. Our results show that alpha-MSH binding is restricted to mammotropes and that a specific subpopulation of these express functional alpha-MSH receptors that are coupled to a Ca2+ signaling pathway. Moreover, alpha-MSH does not compete with dopamine antagonists/agonists for the same binding site.
Interactions between thyroid hormone and tryptophan transport in rat liver are modulated by thyroid status
Authors: Kemp H. F. , Taylor P. M.
Published: December 2017

We have identified both N-ethylmaleimide (NEM)-resistant (system T) and NEM-sensitive (system L1) L-[3H]tryptophan transporters in sinusoidal membrane vesicles (SMVs) from euthyroid, hypothyroid (propylthiouracil-treated), and hyperthyroid [L-3,5,3'-triiodothyronine (L-T3)-injected] rats. L-[125I]T3 associates with SMVs largely by surface binding. Kinetic characteristics of tryptophan uptake and T3 binding (transporter or receptor abundance and substrate affinity) are not significantly affected by thyroid status. T3 and thyroxine (T4) inhibit NEM-resistant tryptophan uptake in SMVs to an extent dependent on the thyroid status of the donor rat, increasing in the order hypothyroid < euthyroid < hyperthyroid; the inhibitor constant for this inhibition (0.3 microM T3) is equal to the dissociation constant for T3 binding. Both T3 binding and T3 inhibition of tryptophan transport in SMVs are markedly reduced by treatments (Triton X-100 or trypsin) that do not significantly affect vesicle integrity or transport of tryptophan and glucose. T3 and/or T4 transport at the liver-plasma interface may be facilitated by direct interactions between hormone receptors and system T transporter proteins. Modulation of such interactions may be important for control of hepatic T4 and/or T3 turnover and aromatic amino acid metabolism during altered thyroid status.
Progesterone-induced changes in sleep in male subjects
Authors: Friess E. , Tagaya H. , Trachsel L. , Holsboer F. , Rupprecht R.
Published: December 2017

Progesterone administration induces a reduction of the vigilance state in humans during wakefulness. It has been been suggested that this effect is mediated via neuroactive metabolites that interact with the gamma-aminobutyric, acidA (GABAA) receptor complex. To investigate the effects of progesterone administration on the sleep electroencephalogram (EEG) in humans we made polysomnographic recordings, including sleep stage-specific spectral analysis, and concomitantly measured plasma concentrations of progesterone and its GABA-active metabolites 3 alpha-hydroxy-5 alpha-dihydroprogesterone (allopregnanolone) and 3 alpha-hydroxy-5 beta-dihydroprogesterone (pregnanolone) in nine healthy male subjects in a double-blind placebo-controlled crossover study. Progesterone administration at 9:30 PM induced a significant increase in the amount of non-rapid eye movement (REM) sleep. The EEG spectral power during non-REM sleep showed a significant decrease in the slow wave frequency range (0.4-4.3 Hz), whereas the spectral power in the higher frequency range (> 15 Hz) tended to be elevated. Some of the observed changes in sleep architecture and sleep-EEG power spectra are similar to those induced by agonistic modulators of the GABAA receptor complex and appear to be mediated in part via the conversion of progesterone into its GABA-active metabolites.
An evaluation of the cross-linking model for the interaction of insulin with its receptor
Authors: Hammond B. J. , Tikerpae J. , Smith G. D.
Published: December 2017

The cross-linking model for insulin receptor interactions, in which a single insulin molecule may form a cross-link between an insulin receptor's alpha-subunits, has been expressed as a formal compartmental model and subjected to a systematic analysis, examining a number of predictions that have been made for this model. The kinetic parameters for the model were obtained by matching data from insulin receptor equilibrium binding studies and rates of formation of the insulin receptor complex. This analytical study has allowed a clear description of the kinetics of the ligand receptor complexes involved in such a mechanism. We conclude that the cross-linking model accounts for the anomaly of the 10-fold concentration difference in high- and low-affinity binding sites found when insulin binding is analyzed by conventional means. However, the phenomenon of acceleration of dissociation of labeled ligand by unlabeled ligand cannot be accounted for as an intrinsic part of the model. We suggest that this phenomenon arises from the destabilization of cross-link formation when a second insulin molecule binds.
Age-related increase in muscle sympathetic nerve activity is associated with abdominal adiposity
Authors: Jones P. P. , Davy K. P. , Alexander S. , Seals D. R.
Published: December 2017

Tonic sympathetic nerve activity (SNA) increases with age, but the mechanisms are unknown. There is evidence that SNA is positively related to total and abdominal body fat, which also increase with age. We tested the hypotheses that 1) the elevation in SNA with age is partially accounted for by higher abdominal and/or total body fat and 2) skeletal muscle is a target of the adiposity-related sympathetic effects. Direct microneurographic recordings of skeletal muscle SNA (MSNA) were obtained during supine rest in 16 older (64 +/- 1 yr, means +/- SE) and 16 young (24 +/- 1 yr) adult males. Central body fat was estimated by waist circumference (WC) and fat mass (FM) by hydrostatic weight. MSNA, WC, and FM were higher in the older vs. young males (44 +/- 2 vs. 22 +/- 2 bursts/min, 91 +/- 2 vs. 79 +/- 1 cm, and 19 +/- 2 vs. 9 +/- 1 kg, respectively; all P < 0.0001). Although univariate correlations were high for MSNA and both WC (r = 0.77) and FM (r = 0.75), stepwise multiple regression analysis revealed WC to be the best predictor of MSNA (R2 = 0.60, P < 0.0001), with FM explaining only an additional 2% of the variance (not significant). Statistically covarying for WC reduced but did not eliminate the difference in adjusted age-group means for MSNA (39 +/- 3 vs. 26 +/- 2 bursts/min, P = 0.003). We conclude that 1) the elevated SNA in older adults is partially related to higher body fat, particularly in the abdominal region, and 2) skeletal muscle is a target of the adiposity-related sympathetic effects observed with aging.
Specific adaptations in muscle and adipose tissue in response to chronic systemic glucose oversupply in rats
Authors: Laybutt D. R. , Chisholm D. J. , Kraegen E. W.
Published: December 2017

Rats minimize hyperglycemia during chronic glucose infusion, but the metabolic processes are unclear. We investigated the tissues involved and the role of altered insulin sensitivity. Cannulated rats were infused with glucose (40 mg.kg-1.min-1) for 1 or 4 days or with saline (control). Hyperglycemia at 1 day (15.3 +/- 1.0 mM) was absent at 4 days (7.5 +/- 0.3 mM), but hyperinsulinemia persisted. Whole body glucose disposal was similarly elevated at 1 and 4 days, implying increased glucose clearance at 4 days (2-fold, P < 0.001). Muscle glucose uptake and glycogen content declined in glucose-infused rats from 1 to 4 days, whereas white adipose tissue (WAT) glucose uptake (6-fold, P < 0.001) and lipogenesis (3-fold, P < 0.001) increased. Muscle and liver triglyceride were doubled at both 1 and 4 days (P < 0.05 vs. control). Insulin sensitivity (assessed during euglycemic clamps) decreased in muscle to 34% of control at 1 and 4 days (P < 0.001 vs. control) and increased fivefold in WAT from 1 to 4 days (P < 0.05). Thus chronic glucose infusion results in a slow increase in efficiency of glucose clearance with enhanced WAT glucose uptake, lipogenesis, and insulin action. In contrast, the adaptation reduces glucose oversupply to muscle. Muscle shows sustained insulin resistance, with lipid accumulation a possible contributing factor.
Prolonged submaximal eccentric exercise is associated with increased levels of plasma IL-6
Authors: Rohde T. , MacLean D. A. , Richter E. A. , Kiens B. , Pedersen B. K.
Published: December 2017

To study the relationship between exercise-related muscle proteolysis and the cytokine response, a prolonged eccentric exercise model of one leg was used. Subjects performed two trials [a branched-chain amino acid (BCAA) supplementation and a control trial]. The release of amino acids from muscle during and after the eccentric exercise was decreased in the BCAA trial, suggesting a suppression of net muscle protein degradation. The plasma concentrations of interleukin (IL)-6 increased from 0.75 +/- 0.19 (preexercise) to 5.02 +/- 0.96 pg/ml (2 h postexercise) in the control trial and in the BCAA supplementation trial from 1.07 +/- 0.41 to 4.15 +/- 1.21 pg/ml. Eccentric exercise had no effect on the concentrations of neutrophils, lymphocytes, CD16+/CD56+, CD4+, CD8+, CD14+/CD38+, lymphocyte proliferative response, or cytotoxic activities. BCAA supplementation reduced the concentration of CD14+/CD38+ cells. This study shows that the concentration of IL-6 in plasma is increased after prolonged eccentric exercise and suggests that the cytokine response is independent of the muscle proteolysis that occur during exercise.
Glucose turnover in humans in the basal state and after intravenous glucose: a comparison of two models
Authors: Overkamp D. , Gautier J. F. , Renn W. , Pickert A. , Scheen A. J. , Schmulling R. M. , Eggstein M. , Lefebvre P. J.
Published: December 2017

This study investigated the ability of two models to represent glucose kinetics in the basal steady state and during an intravenous glucose tolerance test (IVGTT). Six young nonobese male subjects were studied after an overnight fast. Two bolus injections of [U-13C]glucose were given 150 min apart, the first without and the second together with concomitant injection of unlabeled glucose. [3-3H]glucose was constantly infused throughout the study and served to provide an independent means for evaluation of system responses. A linear time-invariant three-compartmental model and the two-compartment time-variant model proposed by Caumo and Cobelli were used to interpret measured time courses of [U-13C]glucose and to reconstruct endogenous glucose production and glucose removal. The ability of the two models to describe the glucose tracer time course was comparable. Simulation studies showed that the two-compartmental time-variant system better predicted measured [3-3H]glucose concentration profiles than did the three-compartmental time-invariant model. However, endogenous glucose production and the integral of excess glucose removal over basal during the IVGTT derived from the two models were almost identical.
Enteral glutamate is the preferential source for mucosal glutathione synthesis in fed piglets
Authors: Reeds P. J. , Burrin D. G. , Stoll B. , Jahoor F. , Wykes L. , Henry J. , Frazer M. E.
Published: December 2017

To measure the source and rate of mucosal glutathione (GSH) synthesis, fed piglets (28 days old; 7.7 kg) received a 6-h infusion of intragastric [U-13C]glutamate (n = 11) either with (n = 5) or without (n = 6) an intragastric infusion of [1-13C]glycine (0-6 h) and [1,2-13C2(U-13C)]glycine (3-6 h). Eighty-four percent of the labeled mucosal GSH-glutamate and 86% of the luminal GSH-glutamate was 13C5. The tracer-to-tracee ratio of GSH-[U-13C]glutamate was 75% of that of mucosal glutamate. Sixty percent of the labeled mucosal glutamate was 13C1, 13C2, or 13C3, but the tracer-to-tracee ratios of these isotopomers in GSH-glutamate were not significantly different from zero. After 3 h of infusion, the tracer-to-tracee ratio of GSH-[U-13C]glycine was 46%, and after 6 h of infusion GSH-[13C1]glycine was 82% of that of mucosal glycine. This suggested that the half-life of mucosal GSH was 2.7 +/- 0.1 h. We concluded that, in fed piglets, mucosal GSH-glutamate derived largely from the direct metabolism of enteral glutamate rather than from glutamate that was metabolized within the mucosa.