bluuebunny/crossref_metadata_2025 huggingface dataset:
Dissociation in plasma renin and adrenal ANG II and aldosterone responses to sodium restriction in rats
Authors: Menachery A. , Braley L. M. , Kifor I. , Gleason R. , Williams G. H.
Published: December 2017

In rats, plasma renin activity (PRA) increases sharply, reaching a plateau within hours of sodium restriction. Plasma aldosterone increases gradually, not reaching a plateau for 1-2 days. To determine whether this dissociation is secondary to the time needed to modify adrenal sensitivity to angiotensin II (ANG II) and to assess the role of locally produced ANG II in this process, rats were salt restricted for 0-120 h. Plasma hormone levels were assessed, adrenal ANG II was measured, and basal and ANG II (1 x 10(-8) M)-stimulated steroidogenesis were determined in vitro. Although PRA attained an elevated plateau within 8 h, plasma aldosterone did not peak until after 48 h of sodium depletion. The in vitro aldosterone sensitivity to exogenous ANG II was not apparent until rats had been salt restricted for 16 h. A plateau (4-fold increase above the ANG II response on high salt) was achieved between 24 and 48 h. Adrenal ANG II also exhibited a similar delayed response that correlates significantly with changes in aldosterone biosynthesis and late pathway activity. Thus the dissociation between PRA and plasma aldosterone may be secondary to a lag in the zona glomerulosa's (ZG) steroidogenic response to ANG II as well as a parallel lag in tissue ANG II production, suggesting that changes in tissue ANG II may mediate ZG sensitivity to ANG II during sodium deprivation.
Increased hepatic insulin proreceptor-to-receptor ratio in diabetes: a possible processing defect
Authors: Dardevet D. , Komori K. , Grunfeld C. , Rosenzweig S. A. , Buse M. G.
Published: December 2017

Hepatic insulin proreceptors and receptors were studied in control and in ketotic diabetic rats 2-4 wk after streptozotocin treatment. Solubilized preparations were partially purified by wheat germ agglutinin-agarose (WGA) and lentil lectin agarose (LLA) chromatography to enrich eluates in insulin receptors and proreceptors, respectively. After phosphorylation with [gamma-32P]ATP, an approximately 190-kDa glycoprotein was identified in LLA eluates as the insulin proreceptor, based on insulin dose-dependent tyrosine autophosphorylation, immunoprecipitation with insulin receptor-specific antibodies, and high-mannose glycosylation. Mature approximately 95 kDa phosphorylated beta-subunits were present in both LLA and WGA eluates. LLA also showed phosphorylated partially processed beta-subunits (approximately 85 kDa) and proreceptors (approximately 190 kDa). Proreceptors comprised less than 1% of the total yield of hepatic insulin receptors. The incorporation of 32P into proreceptors (per gram liver or DNA) was 4.7- or 4.5-fold greater in diabetic vs. control rats, whereas receptor labeling increased only 1.8- or 1.5-fold in diabetic rats. beta-Subunit autophosphorylation per receptor was identical in control and diabetic rats. The phosphorylation data suggested a diabetes-associated 2.6-fold increase in proreceptor-to-receptor ratios. When assessed by cross-linking with 125I-labeled insulin or by immunoblotting, proreceptor-to-receptor ratios were increased 1.5- and 3.1-fold, respectively, in diabetic rats. The data suggest that uncontrolled diabetes may alter insulin receptor processing.
Adenosine A1-receptor coupling to phosphoinositide metabolism in pregnant guinea pig myometrium
Authors: Schiemann W. P. , Buxton I. L.
Published: December 2017

Adenosine contracts pregnant and nonpregnant guinea pig myometrial smooth muscle (MSM). We have 1) described dissociation of A1-adenosine receptors from adenylate cyclase inhibition in nonpregnant MSM (M. A. Smith, J. L. Silverstein, D. P. Westfall, and I. L. O. Buxton, Cell. Signal. 1: 357–365, 1989); 2) described appearance of such inhibitory coupling in pregnant MSM [W. P. Schiemann, D. P. Westfall, and I. L. O. Buxton, Am. J. Physiol. 261 (Endocrinol. Metab. 24): E141-E150, 1991]; and 3) demonstrated a role for myometrial A1 receptors in the rapid formation of D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] in nonpregnant MSM and the cyclooxygenase dependence of this effect (W. P. Schiemann, K. O. Doggwiller, and I. L. O. Buxton. J. Pharmacol. Exp. Ther. 258: 429–437, 1991). To further characterize adenosine action in pregnant tissue, we explored A1 coupling to increased phosphoinositide hydrolysis in near-term pregnant MSM. The A1-receptor agonist (+)-N6-(2-phenylisopropyl)adenosine stimulates the rapid dose-dependent formation of Ins(1,4,5)P3 and stimulates rapid degradation of uterine inositol monophosphates (InsP) in a manner paralleling increases in inositol polyphosphates. Both A1-mediated responses were blocked by the A1 antagonist 8-(p-sulfophenyl)theophylline, and, unlike the effect observed in nonpregnant MSM, treatment of pregnant MSM with either meclofenamate or indomethacin failed to block A1-mediated increases in Ins(1,4,5)P3. Pretreatment of MSM with either Li+ or pertussis toxin failed to alter either Ins(1,4,5)P3 formation or InsP degradation. Furthermore, assay of inositol phosphomonoesterase (InsPase) activity in the presence or absence of Li+ confirmed the existence of an MSM Li(+)-insensitive InsPase enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)
Insulin resistance is associated with high sodium-lithium countertransport in essential hypertension
Authors: Doria A. , Fioretto P. , Avogaro A. , Carraro A. , Morocutti A. , Trevisan R. , Frigato F. , Crepaldi G. , Viberti G. , Nosadini R.
Published: December 2017

The nature of the association between essential hypertension and insulin resistance remains unknown. We measured plasma glucose and insulin levels after an oral glucose tolerance test (OGTT), as well as insulin sensitivity (using a euglycemic hyperinsulinemic clamp), glucose turnover (Rd; using [6,6-2H2]- and [3-3H]glucose isotopic dilution), and forearm net balance of glucose (using arterial-venous difference) in 22 hypertensive patients with high (H2) red blood cell (RBC) sodium-lithium countertransport (Na(+)-Li+ CT; greater than 0.41 mmol.l RBC-1.h-1), 21 hypertensive patients with normal (H1) Na(+)-Li+ CT, and 22 normotensive controls (C). After OGTT, H2 patients had higher plasma glucose and insulin levels than H1 and C. During euglycemic hyperinsulinemia (approximately 100 microU/ml) Rd was lower in H2 [21.7 +/- 1.4 (SE) mumol.kg-1.min-1] than in H1 (44.3 +/- 2.9; P less than 0.01) and C (48.1 +/- 3.0; P less than 0.01), and an inverse correlation was found between rates of Na(+)-Li+ CT and Rd in H1 and H2 (rs = -0.76; P less than 0.01). Forearm glucose uptake was 40-50% lower in H2 compared with H1 and C (P less than 0.01). Lactate concentration increased more in C (from 511 +/- 24 to 1,207 +/- 69 microM) and in H1 (from 564 +/- 40 to 1,122 +/- 99) than in H2 (from 581 +/- 42 to 950 +/- 102, P less than 0.05 vs. both). Forearm blood flow increased more in C (31%, P less than 0.05) and H1 (22%, P less than 0.05) than in H2 (12%).(ABSTRACT TRUNCATED AT 250 WORDS)
31P-NMR study of different hypothyroid states in rat leg muscle
Authors: Kaminsky P. , Klein M. , Robin-Lherbier B. , Walker P. , Escanye J. M. , Brunotte F. , Robert J. , Duc M.
Published: December 2017

Using phosphorus nuclear magnetic resonance spectroscopy, this study was undertaken to determine the effects of experimental hypothyroidism on muscle bioenergetics. The peaks of phosphocreatine (PCr), Pi, phosphodiesters (PDE), sugar phosphomonoesters, and ATP were obtained at rest, during a 2-Hz hindleg muscle stimulation, and during a subsequent recovery period from four groups of anesthetized rats as follows: one control and three hypothyroid (HT) groups treated by propylthyouracil during 2, 4, and 6 wk, respectively. Resting spectra showed a significant rise in Pi by 30% and decreased intracellular pH and PCr/Pi in all three HT groups. PDE progressively increased to 200% of its initial value with hypothyroidism duration. Muscle stimulation did not lead to significant differences in PCr depletion. The percentage of PCr recovery is less in HT muscle than in control muscle. An abnormal H+ metabolism is obvious in all three HT groups. These results indicate abnormal bioenergetics in HT muscle and suggest an impairment of mitochondrial metabolism and of the H+ efflux. They also evoke a high sensitivity of cellular energetics to thyroid deficiency.
Contribution of adrenergic mechanisms to glucose counterregulation in humans
Authors: De Feo P. , Perriello G. , Torlone E. , Fanelli C. , Ventura M. M. , Santeusanio F. , Brunetti P. , Gerich J. E. , Bolli G. B.
Published: December 2017

To assess the role of adrenergic mechanisms during prolonged hypoglycemia, eight normal subjects were studied on six occasions. In study 1, insulin was infused subcutaneously (15 mU.m-2.min-1 for 12 h), and plasma glucose concentration (PG) decreased from 89 +/- 2 to 50 +/- 1 mg/dl. In study 2 (insulin as in study 1 + propranolol and phentolamine + variable glucose to maintain PG as in study 1), the rate of hepatic glucose production (HGO, [3-3H]glucose) was approximately 30% lower after 1.5 h, and the rate of peripheral glucose utilization (GU) was approximately 15% greater after 5 h. To quantitate the effects of adrenergic mechanisms on glucose counterregulation, in a control study (study 3), glucoregulatory hormone secretion was blocked, and the hormones were reinfused to reproduce study 1. When alpha- and beta-blockade plus variable glucose were superimposed to study 3 (study 4), HGO was approximately 25% lower (after 2 h), and GU was approximately 10% greater (after 6 h) vs. study 3. When glucose was not infused to match PG of study 3 (study 5), severe hypoglycemia developed (PG at 7 h 36 +/- 2 vs. 62 +/- 3 mg/dl). Finally, when glucose was not infused during alpha- and beta-blockade of study 2 (study 6), PG was 49 +/- 3 mg/dl at 7 h vs. 65 +/- 3 mg/dl of the control study (study 1), despite greater secretion of glucagon, growth hormone, and cortisol. It is concluded that adrenergic mechanisms play a key counterregulatory role, even in the presence of appropriate responses of glucagon and that greater increases in glucagon (and other counterregulatory hormones) cannot compensate fully for absent contribution of adrenergic mechanisms to counterregulation.
Glucose regulation of lipid metabolism in humans
Authors: Carlson M. G. , Snead W. L. , Hill J. O. , Nurjhan N. , Campbell P. J.
Published: December 2017

The role of plasma glucose in the regulation of lipid metabolism in humans, independent of associated changes in hormone concentrations, is controversial. Therefore we examined the role of glucose in the regulation of lipolysis and free fatty acid (FFA) reesterification in six healthy lean male volunteers. Blood glucose concentration was clamped at either 5 or 10 mM during 2-h pancreatic-pituitary clamps. Glycerol and palmitate turnover were measured by isotope dilution ([1-14C]palmitate and [2H5]-glycerol). All hormone concentrations were the same during the euglycemic and hyperglycemic studies. FFA turnover, which represents the difference between lipolysis and FFA reesterification, was reduced 30% by hyperglycemia (29 +/- 2 vs. 20 +/- 3 mumol.kg fat mass-1.min-1, P less than 0.05). Glycerol turnover, which represents lipolysis only, was reduced to a similar extent (9.4 +/- 0.9 vs. 6.2 +/- 0.7 mumol.kg fat mass-1.min-1, P less than 0.05). We conclude that glucose regulates lipid metabolism, independently of changes in hormone concentrations. The equivalent suppression of glycerol and FFA turnover indicates that the effect is mediated by suppression of lipolysis and not by stimulation of FFA reesterification.
Glucocorticoids in the CNS regulate BAT metabolism and plasma insulin in ob/ob mice
Authors: Walker H. C. , Romsos D. R.
Published: December 2017

Adrenalectomy stimulates depressed brown adipose tissue (BAT) metabolism and decreases hyperinsulinemia in ob/ob mice, with minimal effects in lean mice. A single intracerebroventricular injection of dexamethasone (250 ng) into adrenalectomized ob/ob mice completely reversed the effects of adrenalectomy on BAT thermogenesis as assessed by mitochondrial GDP binding, approximately doubled plasma insulin, lowered whole body metabolic rates by 17%, and increased food intake by 19%. These responses were rapid in onset, with changes in BAT metabolism and plasma insulin occurring within 30 min of dexamethasone injection. Adrenalectomized lean mice were much less responsive to dexamethasone than their ob/ob counterparts. The dexamethasone-induced decrease in BAT thermogenesis in adrenalectomized ob/ob mice was associated with an organ-specific decrease in BAT sympathetic nerve activity as assessed by norepinephrine turnover, whereas the dexamethasone-induced increase in plasma insulin was blocked by atropine, suggesting involvement of the parasympathetic nervous system. Intracerebroventricular injection of corticotropin-releasing hormone did not affect BAT thermogenesis in dexamethasone-injected adrenalectomized ob/ob mice but markedly lowered plasma insulin concentrations, possibly by suppression of the parasympathetic nervous system. In conclusion, dexamethasone alters regulation of the autonomic nervous system in ob/ob mice.
Hepatic metabolism of glucose, galactose, and lactate after milk feeding in newborn lambs
Authors: Spedale S. B. , Battaglia F. C. , Sparks J. W.
Published: December 2017

The purpose of this study was to test the ability of the liver to efficiently clear substrates absorbed from the gastrointestinal tract after a feeding before entry into the systemic circulation. We placed a hepatic vein (HV) catheter in utero at 135-140 days gestation. The lamb was then allowed to deliver spontaneously, and additional catheters were placed in the portal vein (PV) and femoral artery (FA) at 1-3 days postnatal age. After at least 2 days recovery, lambs were fasted overnight at 4-10 days of age and then allowed to nurse ad libitum. There was significant hepatic glucose release during fasting and after 80 min postprandially. No net hepatic uptake of glucose was observed before or after feeding. PV galactose was significantly greater than FA and HV from 40 to 160 min after feeding (P less than 0.005). Hepatic glucose extraction was negligible in the fasted state (1 +/- 3%) and increased after feeding to 75 +/- 2% when PV galactose was less than 1.0 mM. There was a significant hepatic arteriovenous concentration difference of lactate (0.23 +/- 0.03 mM) and of oxygen (0.27 +/- 0.01 mM), which did not change significantly after feeding. The metabolic quotient for galactose increased significantly after feeding, such that galactose was the largest carbon contributor for postprandial hepatic carbon accretion. After a milk feeding, the newborn liver efficiently extracts galactose, lactate, and oxygen, but not glucose.
Protein kinase C differentially modulates PTH- and PGE2-sensitive adenylate cyclase in osteoblast-like cells
Authors: Freyaldenhoven A. M. , Gutierrez G. E. , Lifschitz M. D. , Katz M. S.
Published: December 2017

The effects of phorbol 12-myristate 13-acetate (PMA), a known activator of protein kinase C, on receptor-mediated stimulation of adenylate cyclase were evaluated in a rat osteosarcoma cell line (UMR-106) with the osteoblast phenotype. Pretreatment of UMR-106 cells with PMA increased parathyroid hormone (PTH)-stimulated adenylate cyclase activity and inhibited prostaglandin E2 (PGE2)-responsive enzyme activity. In addition, PMA enhanced enzyme activation by forskolin, which is thought to exert a direct stimulatory action on the catalytic subunit of adenylate cyclase. The regulatory effects of PMA were concentration dependent and of rapid onset (less than or equal to 1 min). Treatment with PMA also resulted in translocation of protein kinase C activity from the cytosol to the particulate cell fraction. Pertussis toxin, which attenuates inhibition of adenylate cyclase mediated by the inhibitory guanine nucleotide-binding regulatory protein (Gi), augmented PTH-sensitive adenylate cyclase activity and reduced the incremental increase in PTH response produced by PMA. The results suggest that activation of protein kinase C increases PTH-stimulated adenylate cyclase activity by actions on Gi and/or the catalytic subunit and decreases PGE2 responsiveness by a mechanism involving the PGE2 receptor.
Impact of uncoupling glucose stimulus from secretion on B-cell release and biosynthesis
Authors: Sako Y. , Eizirik D. , Grill V.
Published: December 2017

We studied the impact of a defined degree of long-term hyperglycemia with or without blockade of attendant insulin release on subsequent B-cell secretory responsiveness and biosynthesis. Nondiabetic rats were infused for 48 h with glucose to produce marked hyperglycemia (21.3 +/- 0.5 mmol/l). Comparable levels of hyperglycemia were upheld when additions were made to this protocol. Hyperglycemia increased plasma insulin 12-fold but depressed glucose (27 mmol/l)-induced insulin secretion in vitro (isolated islets) by 67% compared with saline-infused rats. Addition of diazoxide infusion during hyperglycemia completely inhibited the hyperglycemia-induced rise in plasma insulin but enhanced glucose-induced insulin release in vitro eightfold compared with islets from rats infused with glucose alone. Addition of insulin (2 U/day) to the diazoxide plus hyperglycemia protocol inhibited the secretory response to glucose in vitro by 46% (P less than 0.05). Proinsulin biosynthesis was enhanced by 67% in islets from rats infused with glucose alone; this effect was paralleled by a similar increase in preproinsulin mRNA. Diazoxide in vivo did not affect these stimulatory effects of hyperglycemia on insulin biosynthesis; however, insulin infusion in vivo abolished the hyperglycemia-induced increase in proinsulin biosynthesis. We conclude that impairment by hyperglycemia of glucose-induced insulin secretion occurs concomitant with stimulation of biosynthesis. Uncoupling of glucose stimulus from secretion crucially affects subsequent secretory responsiveness but not biosynthesis. Insulin biosynthesis is depressed by direct or indirect effects of circulating insulin.
Insulin receptor kinase activity in muscles and white adipose tissue during course of VMH obesity
Authors: Pujol A. , Cousin B. , Burnol A. F. , Loizeau M. , Picon L. , Girard J. , Penicaud L.
Published: December 2017

Early after lesion of the ventromedial hypothalamus nuclei (VMH), insulin-induced glucose utilization is increased in white adipose tissue (WAT), whereas oxidative and glycolytic muscles are, respectively, normoresponsive or resistant to insulin. Five weeks later, all of the muscles are resistant, whereas WAT returns to normal responsiveness. The aim of this study was to characterize the insulin receptor kinase activity in WAT and muscles 1 and 6 wk after lesion. The number and affinity of insulin receptors were not modified in any of the tissues studied. Autophosphorylation and phosphorylation of an exogenous substrate were similar in oxidative and glycolytic muscles of VMH and control rats both 1 and 6 wk after the lesion. Insulin receptors from WAT of 1-wk VMH rats exhibited a 2.5-fold increase in insulin-stimulated autophosphorylation and phosphorylation. Six weeks after the lesion, both autophosphorylation and phosphorylation returned to normal values. This suggests that insulin receptor tyrosine kinase activity does not play a significant role in the insulin resistance of skeletal muscles but has a crucial role in mediating the variations of insulin action on WAT observed during the development of VMH obesity.
Xanthine oxidase and dehydrogenase activities in rat ovarian tissues
Authors: Margolin Y. , Behrman H. R.
Published: December 2017

The production of reactive oxygen species in the ovary is rapidly inducible, but the nature of the generator is unknown. One possibility is xanthine oxidase (XO), an enzyme that produces superoxide in the presence of hypoxanthine (or xanthine) and oxygen. The objective of the present studies was to measure levels of XO in follicular and luteal tissue to determine whether XO may be a source of reactive oxygen species in the rat ovary. Ovarian levels of XO were about one-fifth of that seen in the liver and adrenal, and XO levels were about one-third of xanthine dehydrogenase (XDH). Preovulatory ovarian levels of XO activity were unchanged after induction of ovulation with gonadotropin and in follicles incubated with gonadotropin. Luteal XO activity was not changed during natural or prostaglandin F2 alpha (PGF2 alpha)-induced luteolysis. Allopurinol, an inhibitor of XO, did not inhibit ovulation or PGF2 alpha-induced luteal regression. Finally, neither catalase and superoxide dismutase nor oxypurinol altered luteal cell function in the presence of hypoxanthine. Thus, while XO is present in the ovary, it does not appear that it is a major source of reactive oxygen species in this organ.
Different roles of IL-1 and TNF on hemodynamics and interorgan amino acid metabolism in awake dogs
Authors: Fukushima R. , Saito H. , Taniwaka K. , Hiramatsu T. , Morioka Y. , Muto T. , Abumrad N. N.
Published: December 2017

The present study examines the effects of interleukin 1 (IL-1) and tumor necrosis factor (TNF) on various hemodynamic parameters and interorgan fluxes of amino acids, glucose, and lactate in chronically catheterized awake dogs. The dogs received 5 micrograms.kg-1.h-1 of either human recombinant IL-1 beta or TNF intravenously for 2 h. Hemodynamic parameters and substrate fluxes across the gut and liver were determined during and 2 h after discontinuation of the cytokine infusions. Substrate fluxes were calculated by blood flows and arteriovenous differences. Both IL-1 and TNF enhanced the uptake of alanine, uptake of lactate, and output of glucose by the liver. These changes were associated with elevated arterial levels of alanine and lactate while arterial levels of glucose decreased. Uptake of glutamine by the liver was reduced by either IL-1 or TNF infusions. The effects of IL-1 and TNF on the hemodynamic parameters and on gut amino acid metabolism varied with the cytokine infused. IL-1 produced hyperdynamic state, increased splanchnic blood flow, and enhanced glutamine uptake by the gut. TNF infusion did not cause a hyperdynamic state, nor did it alter the gut handling of amino acids. We conclude that IL-1 and TNF exert distinct different effects on the systemic and hemodynamic parameters and on interorgan balances of amino acids, glucose, and lactate across the gut and the liver.
24-hour energy expenditure and sympathetic activity in postobese women consuming a high-carbohydrate diet
Authors: Astrup A. , Buemann B. , Christensen N. J. , Madsen J.
Published: December 2017

It has been suggested that the energy expenditure (EE) of formerly obese subjects (postobese) is highly susceptible to the dietary carbohydrate content and that a high dietary carbohydrate-to-fat ratio may increase their 24-h EE. We studied eight obese women before and after weight normalization (postobese state) and compared them with eight matched controls. Twenty-four hour EE, substrate oxidations, and 24-h heart rate were measured in respiratory chambers on a fixed physical program, while the postobese and controls were in macronutrient balance on a high-carbohydrate diet. Mean 24-h EE decreased from the obese to the postobese state (P less than 0.01), but it remained higher in the postobese women than in the controls (8,292 +/- 153 vs. 7,646 +/- 190 kJ/day, P = 0.01). The higher EE in the postobese group was entirely covered by a 22% higher carbohydrate oxidation (P = 0.006). The mean 24-h heart rate, as measured by telemetry, was also higher in the postobese group than in the control group (74 vs. 66 beats/min, P less than 0.03). Plasma norepinephrine (NE) concentrations were greater by 50% in the postobese as compared with the controls (P = 0.004), and differences in plasma NE concentrations entirely accounted for the group difference in 24-h EE and heart rate between postobese and controls, as analyzed by stepwise regression analysis. We conclude that postobese women on a high-carbohydrate-low-fat diet have an enhanced sympathetic nervous system activity, which is responsible for the higher 24-h EE and heart rate. These findings may have implications for understanding the pathophysiology and for the treatment of obesity.
Thermogenesis in obese women: effect of fructose vs. glucose added to a meal
Authors: Schwarz J. M. , Schutz Y. , Piolino V. , Schneider H. , Felber J. P. , Jequier E.
Published: December 2017

To assess the effect of a fructose meal on resting energy expenditure (EE), indirect calorimetry was used in 23 women (10 lean and 13 obese) for 30 min before and 6 h after the ingestion of a mixed meal containing 20% protein, 33% fat, and either 75 g glucose or 75 g fructose as carbohydrate source (47%). Expressed as a percentage of the energy content of the meal, the thermogenic response to the fructose meal was significantly greater (10.2 +/- 0.5%) than that of the glucose meal (8.4 +/- 0.4%, P less than 0.01). This difference was still apparent when the lean and obese women were considered separately. The mean respiratory quotient during the 6-h postprandial period was significantly greater (P less than 0.01) for the fructose (0.85 +/- 0.01) than for the glucose meal (0.83 +/- 0.01) in the combined subjects. In addition, cumulative carbohydrate oxidation was significantly greater after the fructose than after the glucose meal (51.1 +/- 2.3 vs. 40.9 +/- 2.0 g/6 h, respectively, P less than 0.01). Only small changes were observed in postprandial plasma levels of glucose and insulin after the fructose meal, but the plasma levels of lactate increased more with fructose than with the glucose meal. These results suggest that there might be some advantages (higher thermogenesis and carbohydrate oxidations) in using fructose as part of the carbohydrate source in diet of people with obesity and/or insulin resistance.
Effects of glucose and insulin on development of impaired insulin action in muscle
Authors: Hansen B. F. , Hansen S. A. , Ploug T. , Bak J. F. , Richter E. A.
Published: December 2017

Rat hindquarters were perfused for 2 h with either 0, 5, or 25 mM glucose in combination with either 0, 50, or 20,000 microU insulin/ml, whereupon responsiveness of glucose uptake to 20,000 microU insulin/ml and 25 mM glucose was measured. Perfusion with 25 mM glucose and 20,000 microU insulin/ml resulted in an initial glucose uptake of 43.6 +/- 3.9 mumol.g-1.h-1, which decreased to 18.7 +/- 1.6 mumol.g-1.h-1 after 2 h (P less than 0.001). Omission of glucose from the perfusate prevented the decrease in responsiveness, whereas 5 mM glucose caused a lesser decrease (to 28.3 +/- 2.2 mumol.g-1.h-1). At 0 and 50 microU insulin/ml the effects of glucose were present but were less pronounced. The decrease in insulin responsiveness of glucose uptake (55%) was accompanied by a lesser decrease (29%) in muscle glucose transport, whereas glucose transport in muscle membrane vesicles, muscle insulin binding, and insulin receptor tyrosine kinase activity were unchanged. Muscle glycogen synthase activity decreased (P less than 0.005) during perfusion with 25 mM glucose and 20,000 microU insulin/ml but did not decrease during perfusion with no glucose and 20,000 microU insulin/ml. It is concluded that insulin responsiveness of glucose uptake in muscle is decreased by exposure to glucose in a dose-dependent manner and the inhibitory effect of glucose is enhanced by simultaneous insulin exposure. The mechanism behind this insulin resistance could partly be explained by a decrease in muscle membrane glucose transport, possibly caused by changes in intracellular milieu.
Monitoring thromboxane in body fluids: a specific ELISA for 11-dehydrothromboxane B2 using a monoclonal antibody
Authors: Reinke M.
Published: December 2017

11-Dehydrothromboxane B2 (11-DHTxB2) concentrations are believed to reflect levels of the in vivo synthesis and release of thromboxane A2. In a specific enzyme-linked immunosorbent assay (ELISA) a monoclonal antibody (MAB) against 11-DHTxB2 (MAB-1E-DHTBR1) recognizes the acyclic form of 11-DHTxB2, as found in the basic range of pH, with a detection limit of 4.6 pg/sample and a binding affinity of 6.1 x 10(9) l/mol. Negligible cross-reactivity was found for thromboxane B2 (0.05%), 2,3-dinor-thromboxane B2 (0.06%), and prostaglandin D2 (0.08%). Validity of the assay was confirmed by a good correlation between radioimmunoassay and ELISA (r = 0.972). Recovery experiments showed an accuracy of r = 0.982. Measurements of 11-DHTxB2 in human serum and urine samples demonstrated the practical applicability of the MAB in ELISA. With the use of different clotting times, the serum level of 11-DHTxB2 ranged from 0.8-1.3 ng/ml (30 min) to 24.1-47.9 ng/ml (4 h). After administration of aspirin the 11-DHTxB2 level of human urine declined from 3.9-5.4 ng/ml to 0.4-1.6 ng/ml after 6 h.
Kinetics of glucose transport in rat skeletal muscle membrane vesicles: effects of insulin and contractions
Authors: Ploug T. , Galbo H. , Ohkuwa T. , Tranum-Jensen J. , Vinten J.
Published: December 2017

To study the mechanism of acceleration of glucose transport in skeletal muscle after stimulation with insulin and contractions, we isolated a subcellular vesicular membrane fraction, highly enriched in the plasma membrane enzyme K(+)-stimulated p-nitrophenylphosphatase and also enriched in some intracellular membranes. Protein recovery, morphology, lipid content, marker enzyme activities, total intravesicular volume, Western blot quantitation of GLUT-1, and glucose-inhibitable cytochalasin B binding were identical in membrane fractions from control, insulin-stimulated, contraction-stimulated, and insulin- and contraction-stimulated muscle. Time course of D-[3H]glucose entry in membrane vesicles at equilibrium exchange conditions showed that initial rate of transport at 30 mM of glucose was increased 19-fold and that equilibrium distribution space was increased 4-fold in vesicles from maximum stimulated muscle. The effects of insulin and contractions on initial rate of transport as well as on equilibrium distribution space were additive, and stimulation increased the substrate saturability of glucose transport. Furthermore, cytochalasin B binding to membranes prepared by using less centrifugation time than usual showed that, after stimulation with insulin and contractions, at least 35% of the total number of glucose transporters were redistributed from one kind of vesicles to a more slowly sedimenting kind of vesicles, probably reflecting translocation within the membrane preparation from intracellular vesicles to the plasma membrane upon stimulation. In the present membrane preparation the effects of insulin and/or contractions on glucose transport resemble those seen in intact muscle, and the effects are thus not dependent on cellular integrity.(ABSTRACT TRUNCATED AT 250 WORDS)
Increased islet blood flow in obese rats: role of the autonomic nervous system
Authors: Atef N. , Ktorza A. , Picon L. , Penicaud L.
Published: December 2017

Hyperinsulinemia, a main feature of both human and animal obesity, has been demonstrated to be due to both an increased sensitivity to nutrient secretagogues and an impairment of the nervous regulation of insulin secretion. Recent studies have shown that pancreatic islet blood flow increases under conditions associated with an enhanced insulin secretion. The aim of this study was to determine whether or not changes in islet blood flow are present in hyperinsulinemic obese rats. Using the nonradioactive microsphere technique, we were able to show a significantly higher islet blood flow in obese rats either of the Zucker strain or Wistar rats after lesion of the ventromedial hypothalamus than in their respective lean controls. Subdiaphragmatic vagotomy had no significant effect on basal islet blood flow of lean rats, whereas it decreased significantly that of obese Zucker rats. Conversely, clonidine, an alpha 2-adrenergic agonist, induced a higher decrease of islet blood flow in obese than in lean Zucker rats. The injection of an intravenous bolus of glucose (375 mg/kg iv) increased significantly more islet blood flow in obese than in lean Zucker rats. It is concluded that obese rats present an increased pancreatic islet blood flow, which may result, at least in part, from exaggerated parasympathetic activity and lower than normal sympathetic activity. This could participate in the hyperinsulinemia observed in these rats.
Evaluation of four mathematical models of glucose and insulin dynamics with analysis of effects of age and obesity
Authors: Walton C. , Godsland I. F. , Proudler A. J. , Felton C. , Wynn V.
Published: December 2017

Despite there being a number of mathematical models of glucose and insulin dynamics, there have been no evaluations of their operation in large groups of subjects. We have carried out intravenous glucose tolerance tests on a group of 182 healthy males, with determination of plasma glucose, insulin, and C-peptide concentrations. Parameters of glucose and insulin dynamics were determined using the minimal model of glucose disappearance, a minimal model of peripheral insulin delivery, and two different models of pancreatic insulin secretion (models I and II). Successful identifications were obtained in 96, 95, 76, and 100% of cases, respectively. The models were evaluated in terms of their ability to recover effects of obesity and aging on carbohydrate metabolism. The glucose disappearance model successfully detected the insulin resistance of both obesity and aging, whereas the peripheral insulin delivery model indicated an increased responsiveness of insulin delivery to glucose in obesity but detected no significant change associated with age. No parameter of pancreatic secretion model I exhibited a significant association with either age or obesity. Insulin secretion model II indicated that the hyperinsulinemia accompanying obesity resulted from both increased pancreatic secretion and decreased hepatic insulin uptake.
Insulin receptor function is preserved in a physiological state of hypoinsulinemia and insulin resistance
Authors: Maury J. , Burnol A. F. , Loizeau M. , Issad T. , Girard J. , Ferre P.
Published: December 2017

The suckling period in the rat is characterized by a continuously low plasma insulin concentration and a physiological insulin resistance, particularly in the adipose tissue. This insulin resistance disappears after weaning on the high-carbohydrate adult diet. We have studied the number, structure, and function of adipose tissue insulin receptors during the suckling-weaning transition. The insulin receptor number determined either on intact adipocytes or after partial purification was higher during suckling (15 days), whereas the affinity was similar when compared with weaned rats (30 days). The molecular weight of the alpha- and beta-subunits were identical in both groups and, when analyzed in nonreducing conditions, the alpha 2 beta 2-form was the unique detectable form of the receptor. Neither the basal and insulin-stimulated autophosphorylation of the insulin receptor beta-subunit nor the tyrosine kinase activity toward a synthetic substrate was decreased during the suckling period. Thus, in the adipose tissue of the suckling rat, a marked insulin resistance is concomitant with a normal insulin receptor number and function.
Effects of infused amino acids on glucose production and utilization in healthy human subjects
Authors: Tappy L. , Acheson K. , Normand S. , Schneeberger D. , Thelin A. , Pachiaudi C. , Riou J. P. , Jequier E.
Published: December 2017

Amino acids have been reported to increase endogenous glucose production in normal human subjects during hyperinsulinemia: however, controversy exists as to whether insulin-mediated glucose disposal is inhibited under these conditions. The effect of an amino acid infusion on glucose oxidation rate has so far not been determined. Substrate oxidation rates, endogenous glucose production, and [13C]glucose synthesis from [13C]bicarbonate were measured in six normal human subjects during sequential infusions of exogenous glucose and exogenous glucose with (n = 5) or without (n = 5) exogenous amino acids. Amino acids increased endogenous glucose production by 84% and [13C]glucose synthesis by 235%. Glucose oxidation estimated from indirect calorimetry decreased slightly after amino acids, but glucose oxidation estimated from [13C]glucose-13CO2 data was increased by 14%. It is concluded that gluconeogenesis is the major pathway of amino acid degradation. During amino acid administration, indirect calorimetry underestimates the true rate of glucose oxidation, whereas glucose oxidation calculated from the 13C enrichment of expired CO2 during [U-13C]glucose infusion does not. A slight stimulation of glucose oxidation during amino acid infusion, concomitant with an increased plasma insulin concentration, indicates that amino acids do not inhibit glucose oxidation.
Regulation of glycogen metabolism in canine myocardium: effects of insulin and epinephrine in vivo
Authors: Laughlin M. R. , Taylor J. F. , Chesnick A. S. , Balaban R. S.
Published: December 2017

Myocardial glycogen synthesis and glucose, lactate, and oxygen extraction were measured in the hearts of anesthetized dogs during infusions of insulin and epinephrine. Glycogen was monitored in vivo using 13C-nuclear magnetic resonance during an infusion of [1-13C]glucose into the left anterior descending artery. Glycogen synthesis was observed during a venous infusion of insulin (1.8 microU.min-1.kg-1), and this newly synthesized glycogen was neither broken down nor was more glycogen synthesized during a subsequent epinephrine infusion (0.5 micrograms.min-1.kg-1). During recovery from epinephrine, glycogen synthesis occurred at 2.1 times the rate seen in the control period. Glycogen synthesis was not stimulated in the absence of epinephrine by control infusions of saline. Glucose uptake was increased by insulin during the control period (from 0.09 to 0.39 mumol.min-1.g-1), so that the combined extraction of glucose and lactate exceeded the requirement for oxidizable substrate calculated from oxygen consumption. The "excess" glucose (0.15 mumol.min-1.g wet wt-1) is presumably available for glycogen synthesis. During recovery from epinephrine, lactate uptake was increased over threefold. Because this additional lactate supplies most of the fuel required for oxidation, the excess glucose available for glycogen synthesis during this period was two times that seen before epinephrine (an average of 0.32 mumol.min-1.g wet wt-1 between 20 and 40 min postepinephrine). These data are consistent with the notion that glycogen synthesis can be activated in the heart without an accompanying increase in glucose uptake by providing an alternative substrate (i.e., lactate) for oxidation.
Thermogenic and hormonal responses to amino acid infusion in septic humans
Authors: Arnold J. , Leinhardt D. , Carlson G. , Gray P. , Little R. A. , Irving M. H.
Published: December 2017

Metabolic effects of a commercially available amino acid infusate were investigated in five preoperative patients with abdominal sepsis and five healthy subjects. Oxygen consumption (VO2) was measured continuously during the 3-h study, and blood samples were taken regularly for hormone and metabolite analyses. During 1 h of preinfusion measurements, VO2 was 15% higher (P less than 0.05) in the septic patients. Preinfusion plasma cortisol, glucagon, and catecholamines were also significantly elevated in the septic group. The amino acid solution (9 g nitrogen; 950 kJ; 227 kcal) was infused into each subject through their central venous catheter during the 2nd and 3rd h of the study. VO2 increased similarly in both groups by approximately 21% during the infusion (P less than 0.05), whereas respiratory quotient increased significantly in only the controls (P less than 0.05). Plasma insulin and glucagon concentrations rose significantly in both groups during the infusion, despite little change in glucose levels. Plasma norepinephrine increased in both groups, although the response was significant in only the control subjects. In summary, the amino acid infusate stimulated metabolic rate similarly in the septic and nonseptic subjects.
Alloxan, but not streptozotocin, increases blood perfusion of pancreatic islets in rats
Authors: Jansson L. , Sandler S.
Published: December 2017

It has recently been shown that selective B-cell toxins alloxan and streptozotocin (STZ) possess marked effects also on the vascular system. To evaluate to what extent changes in blood perfusion of islets induced by alloxan or STZ could be of importance for diabetogenic action of these compounds, we first investigated acute effects of alloxan (75 mg/kg body wt iv) and STZ (40 mg/kg body wt iv) on both whole pancreatic blood flow (PBF) and islet blood flow (IBF) in adult rats. Alloxan caused a marked increase in IBF, which was most pronounced 3 min after administration and remained for 30 min. PBF, however, was decreased 3 min after alloxan administration but was similar to that of control animals from 10 min and onward. These two opposite effects on IBF and PBF caused the fraction of whole PBF diverted through islets to increase from approximately 10 to 50%. Pretreatment with glucose (2 g/kg body wt iv), indomethacin (3.5 mg/kg body wt iv), dimethyl sulfoxide (10 ml/kg body wt ip of a 33% solution), superoxide dismutase (SOD, 1,000 kU/kg body wt iv), NG-methyl-L-arginine (30 mg/kg body wt iv), theophylline (7 mg/kg body wt iv), or terbutaline (1 mg/kg body wt iv) failed to affect stimulation of IBF by alloxan observed at 3 min. SOD was found to exert a marked stimulation of IBF both when given alone and together with alloxan. Alloxan increased IBF and decreased PBF also in a syngeneic pancreaticoduodenal graft in rats but did not affect flow distribution in a perfused pancreas-duodenum preparation.(ABSTRACT TRUNCATED AT 250 WORDS)
Effects of fish and safflower oil feeding on subcellular glucose transporter distributions in rat adipocytes
Authors: Ezaki O. , Tsuji E. , Momomura K. , Kasuga M. , Itakura H.
Published: December 2017

Effects of fish oil feeding on glucose transport systems and cell size in rat adipocytes were examined and compared with those of safflower oil or carbohydrate feeding under isoenergy intake conditions. Glucose transport activity was assessed by measuring 3-O-methyl-D-glucose transport. The concentration of erythrocyte type glucose transporter (GLUT-1) and muscle/fat type transporter (GLUT-4) was measured by immunoblotting. The amount of each transporter in intact cells was estimated by the amount of transporter and protein of each membrane fraction and by the recovery of marker enzymes. In cells from safflower-fed rats compared with those from carbohydrate-fed rats, insulin-stimulated glucose transport activity per cell decreased to 51% after a 1-wk feeding, and cell size increase became larger with these effects and continued for at least 4 wk. At 1 wk of feeding, GLUT-1 and GLUT-4 per cell in plasma membrane from insulin-treated cells decreased to 62 and 35%, respectively, with concomitant transporter decreases in the low-density microsome fraction. In cells from high-fish oil-fed rats in which two-thirds of safflower oil was replaced by fish oil, when compared with those from safflower oil-fed rats, insulin-stimulated glucose transport activity increased 1.7-fold after 1 wk of feeding with concomitant cellular GLUT-1 and GLUT-4 increases, but its effect declined thereafter. Parallel with this time course, cell size increase was smaller after 1 wk, but this effect also declined thereafter.(ABSTRACT TRUNCATED AT 250 WORDS)
Amylin and insulin in rat soleus muscle: dose responses for cosecreted noncompetitive antagonists
Authors: Young A. A. , Gedulin B. , Wolfe-Lopez D. , Greene H. E. , Rink T. J. , Cooper G. J.
Published: December 2017

Increasing concentrations of amylin progressively depressed the maximal insulin-stimulated radioglucose incorporation into soleus muscle glycogen, but did not substantively change the EC50 (range 0.78 to 1.52 nM); these findings show noncompetitive, insurmountable antagonism of insulin action by amylin. The results from 36 combinations of different insulin and amylin concentrations were used to construct a response surface that can be used to predict the response for any combination of insulin and amylin concentration. The predicted response to a constant ratio of insulin and amylin concentration is a bell-shaped curve. The experimentally determined response to increasing amounts of an amylin-insulin mixture (molar ratio of 0.14:1, within the range measured for pancreatic secretion and plasma levels) gave a bell-shaped response rather than the sigmoidal response seen with insulin alone. The amylin dose-response relation in the soleus system provides a useful bioassay for amylin agonists. The dose response for highly purified, synthetic human amylin obtained by measuring amylin concentrations by radioimmunoassay in the incubation medium gave an EC50 of 456 pM (+/- 0.18 log units). Human amylin had a potency greater than or equal to that of human insulin in this highly insulin-sensitive preparation.
Differential effects of interleukin-1 and tumor necrosis factor on ketogenesis
Authors: Memon R. A. , Feingold K. R. , Moser A. H. , Doerrler W. , Adi S. , Dinarello C. A. , Grunfeld C.
Published: December 2017

To determine the role of cytokines in mediating the decrease in ketones associated with infection, we studied the effect of endotoxin (LPS), interleukin-1 (IL-1), and tumor necrosis factor (TNF) on serum and hepatic ketone body levels (KB), serum free fatty acids (FFA), and hepatic malonyl-CoA levels. LPS decreased serum and hepatic KB in C57Bl/6 (LPS sensitive) mice, whereas it had little effect in C3H/HeJ (LPS resistant) mice, whose macrophages lack the ability to produce IL-1 and TNF in response to LPS, suggesting that IL-1 and TNF may mediate this effect. IL-1 and TNF decreased serum KB in both strains of mice. As seen with LPS, IL-1 decreased hepatic KB, whereas TNF had no such effect. LPS, IL-1, and TNF increased hepatic malonyl-CoA levels. TNF acutely raised serum FFA, whereas LPS and IL-1 did not. Postulating that the TNF-induced increase in FFA overrides the inhibitory effect of malonyl-CoA on fatty acid oxidation and ketogenesis, we used R-2-phenylisopropyladenosine to block TNF-induced lipolysis and demonstrated that in the absence of increased fatty acid flux, TNF inhibits KB formation. As seen with LPS, IL-1, but not TNF, decreased KB in the fasting state. These data suggest that IL-1 and TNF may mediate the antiketogenic effect of infection and that IL-1 has properties closest to that of LPS.
Insulin-regulated sorting of glucose transporters in 3T3-L1 adipocytes
Authors: Robinson L. J. , James D. E.
Published: December 2017

Two glucose transporters (GLUT-4 and GLUT-1) move from within the cell to the plasma membrane (PM) when 3T3-L1 adipocytes are stimulated with insulin. To study the sorting of these two molecules, vesicles containing GLUT-4 and GLUT-1 were immunoadsorbed from basal and insulin-treated cells. Two different vesicle populations were isolated as follows: 1) a compartment that contained the majority of intracellular GLUT-4 and GLUT-1 and 2) a subpopulation of vesicles containing 43% of the intracellular GLUT-4 that was highly insulin regulatable and that contained relatively low levels of GLUT-1. After incubation at 19 degrees C, basal glucose transport was slightly increased, whereas insulin-dependent transport was blocked. Consistent with these observations, cell surface GLUT-1 levels were increased in the basal state, whereas insulin-dependent translocation of GLUT-4 to the PM was blocked at 19 degrees C. However, insulin-dependent sorting of GLUT-4 within the intracellular compartment was still evident at 19 degrees C. These data indicate that GLUT-1 and GLUT-4 are heterogeneously distributed throughout the same intracellular compartment in 3T3-L1 adipocytes. Furthermore, we have uncoupled two distinct steps in the insulin-dependent movement of GLUT-4 to the cell surface. These include movement of GLUT-4 out of its storage compartment and accumulation of GLUT-4 at the cell surface. Only the former step occurs in cells preincubated at 19 degrees C.