bluuebunny/crossref_metadata_2025 huggingface dataset:
Acute metabolic effects of adrenergic agents in swine
Authors: Mersmann H. J.
Published: December 2017

A pig model in vivo was used to confirm the unique specificity for stimulation of porcine adipose tissue lipolysis by norepinephrine analogues in vitro. Plasma free fatty acid and blood glycerol concentrations were monitored as probable indicators of adipose tissue lipolysis. Plasma glucose and lactate concentrations, blood pressure, and heart rate were monitored also. Norepinephrine analogues were infused intravenously. Several compounds, classified as either beta 1- or beta 2-adrenergic agonists, that stimulated lipolysis in vitro also increased plasma free fatty acid and blood glycerol concentrations in vivo. Tazolol (beta 1) and quinterenol (beta 2) did not stimulate lipolysis in vitro and likewise did not elevate plasma free fatty acid or blood glycerol concentrations in vivo. Clenbuterol and zinterol did not stimulate lipolysis in vitro but elevated plasma free fatty acid concentrations in vivo, implying indirect effects. Isoproterenol stimulation of plasma free fatty acid and blood glycerol concentrations in vivo was antagonized by propranolol, implying the beta-adrenergic nature of the receptors. Infusion of purported beta 1- and beta 2-adrenergic antagonists suggested control of lipolysis in vivo predominantly by beta 1-adrenergic receptors; however, because the results in vitro do not indicate this specificity, differential pharmacodynamics of the antagonists are suggested rather than designation of receptor subtypes. There was no evidence for alpha-adrenergic mediated inhibition of adipose tissue lipolysis in vivo, confirming observations in vitro.
Relationship between plasma and platelet epinephrine concentrations in humans
Authors: Rosen S. G. , Sanfield J. A. , Morrow L. A. , Zweifler A. J.
Published: December 2017

Nine healthy humans received sequential 80-min infusions of saline and epinephrine (EPI) at 0.55 and 2.75 micrograms X min-1 X m-2 to determine whether increases in arterialized venous plasma EPI within the physiological range affect platelet EPI levels. The platelet EPI concentration was significantly higher than basal levels only during the 2.75 micrograms X min-1 X m-2 EPI infusion (P less than 0.02). To assess the role of platelets in EPI clearance from plasma, four human subjects underwent a 2.75 micrograms X min-1 X m-2 EPI infusion during beta-adrenergic blockade with propranolol. Propranolol decreased plasma EPI clearance (P less than 0.01) and increased both the plasma and platelet EPI concentrations (P less than 0.01) during the EPI infusion. To determine the changes in plasma and platelet EPI during and after EPI infusion, seven subjects received an 80-min EPI infusion at 2.75 micrograms X min-1 X m-2. After discontinuation of the EPI infusion, plasma EPI levels returned to base line within 30 min, whereas platelet EPI levels were significantly elevated above base line for 120 min (P less than 0.02 vs. base line). Platelets concentrate EPI during increases in plasma EPI, which are similar to the plasma levels seen in physiological and pathophysiological states in humans. The accumulation of catecholamines by platelets may be an additional mechanism in the regulation of plasma catecholamine levels in humans.
Protein synthesis during hypoxia in fetal lambs
Authors: Milley J. R.
Published: December 2017

The goal of this study was to test the hypothesis that the rate of fetal protein synthesis decreases during fetal hypoxia. Catheters were inserted into 13 sheep fetuses under maternal spinal and local fetal anesthesia. Five days after surgery, an infusion of L-[1-14C]tyrosine (0.05-0.25 muCi/min) was begun. Measurements were first made when tyrosine-specific activity reached steady state at 3 h and then again 3 h after fetal oxygen delivery was reduced by lowering the maternal inspired oxygen concentration. Fetal tyrosine uptake across the umbilical circulation was 1.34 +/- 0.20 mumol X kg-1 X min-1 during normoxia and decreased to 0.72 +/- 0.12 mumol X kg-1 X min-1 during hypoxia (P = 0.017). Tyrosine use from the plasma compartment was 1.25 +/- 0.18 mumol X kg-1 X min-1 during normoxia and decreased to 0.64 +/- 0.12 mumol X kg-1 X min-1 (P = 0.0005) during hypoxia. Fetal tyrosine use decreased during reduced oxygen delivery because the rate of tyrosine use for fetal protein synthesis decreased from 0.97 +/- 0.17 to 0.38 +/- 0.09 mumol X kg-1 X min-1 (P = 0.0004). Fetal oxygen consumption decreased by 40 mumol X kg-1 X min-1 during hypoxia. Decreased protein synthesis reduced the energy needed for protein synthesis and explained 28 mumol X kg-1 X min-1 of this reduction.
Type I receptors in parotid, colon, and pituitary are aldosterone selective in vivo
Authors: Sheppard K. , Funder J. W.
Published: December 2017

Previous in vivo studies have demonstrated that type I receptors in the rat kidney are aldosterone selective, whereas those in the hippocampus do not appear to discriminate between aldosterone and corticosterone. We have injected mature rats with [3H]aldosterone or [3H]corticosterone plus 100-fold excess of RU 28362, with or without unlabeled aldosterone or corticosterone, and compared type I receptor occupancy in two classic mineralocorticoid target tissues (parotid and colon) and in the pituitary. Mature rats were killed 10-180 min after tracer administration; [3H]aldosterone was well taken up and retained in all tissues, whereas [3H]corticosterone was significantly retained only in the pituitary 10 min after tracer administration. To assess a possible role for corticosterone-binding globulin (CBG) in conferring aldosterone specificity on type I receptors, 10-day-old rats (with very low levels of CBG) were similarly injected. In the colon and parotid, [3H]aldosterone binding was at least an order of magnitude higher than that of corticosterone; in the pituitary aldosterone binding was approximately three times that of corticosterone. We interpret these data as evidence that in the parotid and colon type I receptors are aldosterone selective by a non-CBG-requiring mechanism, whereas in the pituitary there appear to be both aldosterone-selective and nonselective type I sites.
Acute effects of ethanol and acetate on glucose kinetics in normal subjects
Authors: Yki-Jarvinen H. , Koivisto V. A. , Ylikahri R. , Taskinen M. R.
Published: December 2017

We compared the effects of two ethanol doses on glucose kinetics and assessed the role of acetate as a mediator of ethanol-induced insulin resistance. Ten normal males were studied on four occasions, during which either a low (blood ethanol 4 +/- 1 mmol/l) or moderate (14 +/- 1 mmol/l) ethanol, acetate, or saline dose was administered. Both ethanol doses similarly inhibited (0.4-0.5 mg.kg-1.min-1, P less than 0.01) basal glucose production. The decrease in Ra was matched by a comparable decrease in glucose utilization (Rd), resulting in maintenance of normoglycemia. During hyperinsulinemia (approximately 70 microU/ml), glucose disposal was lower (1.2-1.7 mg.kg-1.min-1, P less than 0.01) in the moderate than the low-dose ethanol or saline studies. During acetate infusion, the blood acetate level was comparable with those in the ethanol studies. Acetate had no effect on glucose kinetics. In conclusion, 1) in overnight fasted subjects, ethanol does not cause hypoglycemia because its inhibitory effect on Ra is counterbalanced by equal inhibition of Rd;2) basal Ra and Rd are maximally inhibited already by small ethanol doses, whereas inhibition of insulin-stimulated glucose disposal requires a moderate ethanol dose; and 3) acetate is not the mediator of ethanol-induced insulin resistance.
Evidence that diminished pituitary responsivity to GHRF is secondary to intracellular GH pool depletion
Authors: Richardson S. B. , Twente S.
Published: December 2017

In a perifused dispersed rat anterior pituitary cell system, growth hormone (GH) secretion became attenuated in response to repeated pulsatile or prolonged exposure to submaximal stimulatory concentrations of rat growth hormone-releasing factor (GHRF). However, persistent intracellular GH stores could be released upon subsequent challenge with the membrane depolarizing agent KCl, forskolin, or the phorbol ester, tetradecanoylphorbol acetate (TPA). The GH secretory response to repeated pulsatile administration of either KCl or forskolin also became attenuated. In these experiments, persistent intracellular GH stores could be released upon subsequent GHRF stimulation. Repeated challenge with pulses of TPA failed to elicit any GH release after the initial stimulatory response, although a subsequent GHRF pulse was stimulatory, indicating persistence of intracellular GH stores. These data are compatible with the hypothesis that the decreased GH secretory responsivity to GHRF, which was observed in the course of these experiments, is caused by the functional depletion of specific secretagogue-sensitive pools of intracellular GH, rather than by receptor-mediated desensitization.
Regional differences in adipocyte lactate production from glucose
Authors: Newby F. D. , Sykes M. N. , DiGirolamo M.
Published: December 2017

Having shown that lactate is an important product of glucose metabolism by rat epididymal adipocytes, we investigated possible regional differences in adipocyte lactate production and the role of the animals' nutritional state and stage of development. [U-14C]glucose metabolism, lactate production, and response to insulin were measured in fat cells isolated from four adipose regions (subcutaneous, epididymal, retroperitoneal, and mesenteric) from young lean and older fatter rats, killed either in the fed state or after fasting for 48 h. In the absence of insulin, mesenteric fat cells from either age group metabolized significantly more glucose per cell and converted more glucose to lactate than cells from other depots, regardless of nutritional state. Adipocytes from fasted lean rats showed a significant increase (approximately 30%) in the relative glucose conversion to lactate in all depots when compared with cells from fed lean rats. Fasting of older fatter rats, however, had limited effects on the relative adipocyte glucose conversion to lactate since lactate production was already high (40-50% of glucose metabolized). Mesenteric fat cells had the lowest relative response to insulin, possibly due to the high basal rate of glucose metabolism. These findings indicate that differences exist among adipose regions in the rates of glucose metabolism, lactate production and response to insulin. The anatomical location of the mesenteric adipose depot, coupled with a high metabolic rate and blood perfusion, suggests that mesenteric adipocytes may provide a unique and more direct contribution of metabolic substrates for hepatic metabolism than adipocytes from other depots.
Exercise-induced fall in insulin and hepatic carbohydrate metabolism during muscular work
Authors: Wasserman D. H. , Williams P. E. , Lacy D. B. , Goldstein R. E. , Cherrington A. D.
Published: December 2017

To examine the role of the exercise-induced fall in insulin, dogs were studied during 150 min of treadmill exercise alone (C) or with insulin clamped at basal levels by an intraportal infusion so as to prevent the normal fall in its concentration (IC). To counteract the suppressive effect of insulin on glucagon release, glucagon was replaced intraportally in a separate group of dogs in which insulin levels were clamped (IC + G). In all dogs, catheters were placed in an artery and in the portal and hepatic veins for sampling and in the vena cava and the portal vein for infusion purposes. Glucose production (Ra) and gluconeogenesis were assessed with isotope and arteriovenous difference techniques. In C, insulin fell 5 +/- 2 microU/ml by the end of exercise and was unchanged in IC (delta 0 +/- 2 microU/ml) and IC + G (delta 0 +/- 1 microU/ml). Glucagon rose 54 +/- 11 pg/ml with exercise in C and was unchanged in IC (delta - 4 +/- 11 pg/ml), and normal increments were restored in IC + G (delta 55 +/- 10 pg/ml). Catecholamines and cortisol rose similarly in all groups. Ra increased by an average of 4.0 +/- 0.4, 0.9 +/- 0.3, and 1.8 +/- 0.4 mg.kg-1.min-1 during exercise in C, IC, and IC + G, respectively. Gluconeogenesis from alanine rose by 212 +/- 34, 91 +/- 39, and 184 +/- 47% with exercise in C, IC, and IC + G.(ABSTRACT TRUNCATED AT 250 WORDS)
Effect of galanin on glucose-, arginine-, or potassium-stimulated insulin release
Authors: Yoshimura T. , Ishizuka J. , Greeley G. H. , Thompson J. C.
Published: December 2017

We have examined the effect of galanin infusion on glucose-stimulated release of insulin from the isolated perfused pancreas of the rat to better characterize the effect of galanin on the first and second phases of insulin release. The effects of galanin on insulin release stimulated by L-arginine or high concentrations of potassium were also examined. When perfusion of galanin was started 4 min before the start of perfusion of high glucose (16.7 mM), galanin (10(-8)-10(-11) M) inhibited both the first and second phases of insulin release in a dose-dependent manner. When perfusion of galanin (10(-8) or 10(-9) M) was started simultaneously with high glucose (16.7 mM), only the second phase of insulin release was suppressed (P less than 0.05). Galanin (10(-9) M) failed to inhibit insulin release stimulated by L-arginine (10 and 5 mM) or potassium (25 and 20 mM). These findings suggest that the inhibitory action of galanin on glucose-stimulated insulin release is exerted on early intracellular events that occur during the stimulation of insulin release and that are common to both phases. Because galanin does not inhibit insulin release stimulated by L-arginine or potassium, galanin may inhibit glucose-stimulated closure of potassium channels.
Testing of the assumptions made in estimating the extent of futile cycling
Authors: Wajngot A. , Chandramouli V. , Schumann W. C. , Kumaran K. , Efendic S. , Landau B. R.
Published: December 2017

In estimating glucose and fructose 6-phosphate futile cycling in vivo, complete detritiation of [2-3H]glucose is assumed at the glucose 6-phosphate level, [3-3H]glucose at triose phosphate formation, and [6-3H]glucose in its conversion to glucose via pyruvate. [3-3H]glucose detritiation via the pentose cycle is assumed to be negligible. Normal and non-insulin-dependent diabetic subjects, in the basal state and infused with glucose, were given [2-3H,2-14C]galactose, and 3H-to-14C ratios in blood glucose were determined. [2-3H,2-14C]glucose was given with acetaminophen, and 3H/14C in urinary glucuronide was determined. Detritiation at glucose 6-phosphate was approximately 80%. [3-3H,1-14C]fructose was infused, and 3H/14C was determined in blood glucose and urinary glucuronide. At triose phosphate, 75-90% of the 3H was removed. The pentose cycle contribution was only a few percent. [6-3H,6-14C]glucose was infused, and 3H/14C in blood lactate was determined. [3-3H,3-14C]lactate was infused, and ratios in blood glucose were determined. Maximally, 10% of 3H from [6-3H]glucose was retained. If glucose and galactose are metabolized in the same hepatic site(s), glucose conversion to three-carbon intermediates in the indirect pathway of glycogen formation occurs in extrahepatic tissue(s). Reported estimates of futile cycling, although qualitatively correct, quantitatively require correction.
Changes of sympathetic nerve activity induced by 2-deoxy-D-glucose infusion in humans
Authors: Fagius J. , Berne C.
Published: December 2017

Microelectrode recording of sympathetic signals in the peroneal nerve was performed in 14 healthy volunteers following infusion of 2-deoxy-D-glucose (50 ml/kg body wt). Heart rate, blood pressure, body temperature, hematocrit, and blood levels of glucose, insulin, and catecholamines were monitored. Muscle nerve sympathetic activity (MSA), which is involved in cardiovascular homeostasis, increased significantly from a base-line level of 19.9 +/- 4.5 (mean +/- SE) bursts/min to a peak 30 min after the start of the infusion of 33.1 +/- 5.1 bursts/min. Skin nerve sympathetic activity (SSA), which is a mixture of sudomotor and vasoconstrictor signals, also increased to a peak at 30 min. The impulse pattern of SSA suggested that the increase involved mainly sudomotor activity, with simultaneous inhibition of vasoconstrictor signals. The time courses of MSA and the circulatory responses suggested that the increase in MSA was not a baroreceptor-induced counteraction of the cardiovascular changes during glucopenia. The responses of MSA and SSA were remarkably similar to those observed previously during insulin-induced hypoglycemia. The relationship between changes of sympathetic outflow, glucose levels, and insulin levels in the present study indicates that the effects observed are consequences of central nervous system glucopenia, with insulin playing a minor role. It is concluded that the sympathoadrenal system responds in a markedly differentiated way to glucopenia.
Insulin inhibition of overnight glucose production and gluconeogenesis from lactate in NIDDM
Authors: Yki-Jarvinen H. , Helve E. , Sane T. , Nurjhan N. , Taskinen M. R.
Published: December 2017

Increased gluconeogenesis contributes to fasting hyperglycemia in non-insulin-dependent diabetes mellitus (NIDDM). We examined whether insulin inhibits gluconeogenesis from lactate by altering the fate of lactate and/or by reducing lactate flux. Seven patients with NIDDM (age 51 +/- 4 yr, body mass index 28 +/- 2 kg/m2) were studied before and 3 wk after achieving normoglycemia with evening insulin therapy. Basal glucose production (Ra) and utilization were measured overnight [( 3-3H]glucose infusion from 9 P.M. to 8 A.M.) and lactate turnover and conversion to glucose between 4 and 8 A.M. [( U-14C]lactate infusion) before and after insulin therapy. During insulin therapy, fasting plasma glucose decreased from 188 +/- 13 to 99 +/- 7 mg/dl (P less than 0.001) due to inhibition of glucose Ra from 3.0 +/- 0.1 to 2.2 +/- 0.1 mumol.kg-1.min-1 (P less than 0.005). Plasma free insulin increased from 6 +/- 1 to 11 +/- 1 microU/ml (P less than 0.005). Plasma lactate concentrations (1.1 +/- 0.2 vs. 1.0 +/- 0.1 mmol/l before vs. after insulin therapy) and the lactate turnover rate (15.6 +/- 0.9 vs. 14.2 +/- 0.8 mumol.kg.min) remained unchanged, whereas the amount of glucose formed from lactate decreased from 2.0 +/- 0.1 to 1.4 +/- 0.2 mumol.kg-1.min-1 (P less than 0.02) and the percent of lactate turnover converted to glucose decreased from 26 +/- 1 to 20 +/- 2% (P less than 0.05). We conclude that insulin inhibits overnight glucose Ra from lactate by decreasing the proportion of lactate diverted towards gluconeogenesis rather than by altering lactate availability or total flux.
Insulin, growth hormone, glucose, and fatty acids in gilts selected for rapid vs. slow growth rate
Authors: Norton S. A. , Zavy M. T. , Maxwell C. V. , Buchanan D. S. , Breazile J. E.
Published: December 2017

Twelve-hour plasma profiles of growth hormone (GH), insulin, glucose, and nonesterified free fatty acids (NEFA), as well as GH, insulin, and glucose responses to saline, glucose, arginine, and human pancreatic growth hormone-releasing factor (GRF) infusion were examined in 15 crossbred gilts, selected for rapid vs. slow growth. For experiment 1 GH and insulin patterns differed (P less than 0.05) between the rapid growth line (RGL) and slow growth line (SGL). Mean GH concentrations in SGL and RGL gilts were 4.1 and 3.2 ng/ml, respectively (P less than 0.05). Plasma GH profile area was greater (P less than 0.08) for SGL than RGL gilts. Glucose level, glucose area, and insulin level were higher (P less than 0.05) for RGL than SGL gilts. Plasma NEFA levels and area were greater (P less than 0.05) for SGL gilts. In experiment 2 responses of GH, insulin, and glucose to saline, glucose, arginine, and GRF infusions varied (P less than 0.05) between and within lines relative to control values. These results indicate that selection for growth rate results in concomitant changes in endocrine and metabolic status.
Effects of central and peripheral dopamine antagonism on aldosterone secretion: evidence for adrenal mechanism
Authors: Stern N. , Eggena P. , Chandler W. , Tuck M. L.
Published: December 2017

Both domperidone (DOMP) and metoclopramide (MCP) are D2 receptor antagonists, MCP being a central and peripheral dopamine antagonist, whereas DOMP is exclusively a peripheral antagonist. MCP, but not DOMP, has been shown to stimulate aldosterone production. To elucidate whether aldosterone stimulation by dopamine antagonism is centrally mediated, we injected DOMP (28 micrograms/kg body wt) via a cannula into the third ventricle in Sprague-Dawley rats. Plasma aldosterone and renin concentration were measured before and 15 min after the injection. Centrally administered DOMP resulted in an increment in plasma aldosterone (23.8 +/- 7.4 ng/dl) that was not significantly greater than that induced by vehicle alone (15.8 +/- 4.5 ng/dl). This increase was inhibited by pretreatment with dexamethasone (100 micrograms three times daily) and attenuated by captopril (1 mg/kg ip) but not by L-beta-3,4-dihydroxyphenylalanine (30 mg/kg), thus reflecting a stress effect. Similarly, central administration of MCP (21 micrograms/kg) resulted in a significant rise in plasma aldosterone. This increase, however, was eliminated by pretreatment with dexamethasone and attenuated by captopril. Peripherally administered DOMP (280 micrograms/kg) had no effect on plasma aldosterone. The effect of DOMP and MCP on aldosterone secretion by freshly obtained adrenal capsules was also tested. Angiotensin II and MCP, but not DOMP, induced a dose-dependent increase in aldosterone secretion, with a maximal increment (15.7 +/- 5.8 ng.mg capsular protein-1.10 min-1; 50% increase) with MCP at 10(-7) M (P less than 0.01 compared with controls). Dopamine completely inhibited this MCP-induced rise in aldosterone release.(ABSTRACT TRUNCATED AT 250 WORDS)
Ion microprobe analysis of bone surface elements: effects of 1,25(OH)2D3
Authors: Bushinsky D. A. , Chabala J. M. , Levi-Setti R.
Published: December 2017

When neonatal mouse calvariae are incubated with 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] there is net calcium efflux from the bone into the medium. The effect of this enhanced cell-mediated Ca efflux on the relative concentrations of mineral 23Na, 39K, and 40Ca has not previously been studied. We used an imaging scanning ion microprobe, utilizing secondary ion mass spectrometry, to compare the relative ion concentrations of Na, K, and Ca on the surface, subsurface, and cross-section of cultured bone incubated in the presence of 1,25(OH)2D3 with the ion concentrations in similar regions of bone incubated in unaltered control medium. Changes in mineral ion concentration were correlated with net fluxes of Na, K, and Ca relative to bone. Calvariae incubated in control medium (24 h at pH approximately 7.40) have abundant surface Na and K relative to Ca (Na/Ca, 85 and K/Ca, 68), whereas the subsurface has less Na/Ca (21) and K/Ca (23), and on cross section the ratios of both Na/Ca (2.0) and K/Ca (1.9) decrease further. After incubation with 10(-8) M 1,25(OH)2D3, there is a significant increase in bone surface Na/Ca (154) and K/Ca (141) without a change in these ratios on the subsurface and a small fall in both ratios on cross section. The linear relationship between Na/Ca and K/Ca across the three regions of bone observed in control calvariae did not change with 1,25(OH)2D3 treatment. As determined by flux measurements there is a net efflux of Ca but not Na or K from bone.(ABSTRACT TRUNCATED AT 250 WORDS)
Cholecystokinin receptors and vagal nerves in control of food intake in rats
Authors: Garlicki J. , Konturek P. K. , Majka J. , Kwiecien N. , Konturek S. J.
Published: December 2017

This study was designed to determine the specificity and physiological nature of short-term satiety effects of cholecystokinin (CCK) in rats with intact and transected vagal nerves. Rats with-the gastric fistulas, closed or open, were used for normal feeding or sham feeding of liquid meal offered for 30 min. CCK-8 (0.5-10 nmol/kg) injected intraperitoneally (ip) 15 min before feeding inhibited food intake dose dependently in both normal-fed and sham-fed rats at a minimal inhibitory dose of 1 nmol/kg. CCK-8 at the same doses caused a potent stimulation of pancreatic protein secretion, reaching maximum at a dose of approximately 0.5 nmol/kg. Pretreatment with a potent CCK receptor antagonist, L-364,718 (2.5 mg/kg ip), increased food intake during normal feeding (but not sham feeding) and almost completely blocked the satiety and pancreatic stimulatory effects of CCK. When feeding was preceded by intragastric administration of proteinase inhibitor (Foy-305, 200 mg/kg), food preload, or diversion of bile-pancreatic secretion to the exterior, there was a significant increase in the plasma level of CCK and an inhibition of food intake by about 36, 78, and 25%, respectively. Pretreatment with L-364,718 completely abolished this inhibition by Foy-305 and bile-pancreatic diversion and reduced that caused by food preload. Among other gut peptides given ip (10 nmol/kg) only bombesin reduced food intake, whereas gastrin, secretin, gastric inhibitory polypeptide (GIP), pancreatic polypeptide (PP), and peptide YY (PYY) were ineffective.(ABSTRACT TRUNCATED AT 250 WORDS)
Defective regulation of thyroxine 5'-deiodinase in brown adipose tissue of ob/ob mice
Authors: Kates A. L. , Himms-Hagen J.
Published: December 2017

Genetically obese (ob/ob) and lean (+/?) mice were exposed to a cold (14 degrees C) environment for 1, 3, 6, 12, 16, or 24 h or remained in a warm (28 degrees C) environment. In ob/ob mice the increase in brown adipose tissue (BAT) thermogenesis (mitochondrial GDP binding) was low and the increase in thyroxine 5'-deiodinase (T5'D) was delayed; there was a reduced increase in serum 3,5,3'-triiodothyronine (T3) level and these mice became hypothermic. Content of uncoupling protein (UCP) was low in BAT of obese mice and no cold-induced increase occurred. Adrenalectomy of obese mice before exposure to cold (14 degrees C) improved defective thermogenic response of BAT and thermoregulation, restored to normal the increases in T5'D activity and serum T3 level, and promoted an exaggerated increase in UCP content, detectable after only 6 h. Adrenalectomy of cold-exposed lean mice did not alter thermoregulation, the increase in BAT T5'D activity, or the increase in serum T3 but enhanced the thermogenic response and allowed a higher UCP content in BAT of cold-exposed mice. We conclude that suppression of the cold-induced increase in T5'D activity in BAT can be added to the other known corticosterone-dependent anomalies of the ob/ob mouse. We speculate that lack of the cold-induced increase in T5'D in BAT of the ob/ob mouse prevents the normal participation of T3 in the trophic response of BAT to cold and underlies the abnormality in this response.
Glucose is required to maintain ATP/ADP ratio of isolated bovine cerebral microvessels
Authors: Gaposchkin C. G. , Tornheim K. , Sussman I. , Ruderman N. B. , McCall A. L.
Published: December 2017

Isolated bovine cerebral microvessels (ICMV) were incubated with different metabolic fuels to determine the effect of each of them on microvessel energy state. With no fuel added to the medium, the ATP/ADP generally decreased from initial values of 1.5-3 down to 1-1.5 over 4 h; the ATP content also declined approximately 50%. In contrast, with glucose present, the ATP/ADP increased, and the ATP content was maintained. Pyruvate, beta-hydroxybutyrate, glutamate, and oleate were ineffective; oleate added together with carnitine gave some improvement but less than with glucose. Oxygen consumption by ICMV did not differ appreciably in fuel-free or glucose-containing medium. Addition of an inhibitor of fatty acid oxidation, 2-tetradecylglycidate, depressed the ATP/ADP. These results suggest that ICMV require glycolysis to maintain both their content of ATP and their ATP/ADP. They also suggest that endogenous lipid is an important fuel for isolated microvessels.
Increased growth and protein deposition in rats treated with antibodies to adipocytes
Authors: Panton D. , Futter C. , Kestin S. , Flint D.
Published: December 2017

Young growing rats treated with an antiserum to adipocytes showed marked reductions in several adipose tissue depots but surprisingly demonstrated increased body weight gain. During the first 3 wk after treatment body weight gain increased by 17% with no effect on food intake, whereas during weeks 3-7 body weight gain increased by 40-50% and was accompanied by a 15% increase in food intake. These animals thus exhibited increased food conversion efficiencies (intake/gain) of approximately 15-20% for almost 2 mo. Subsequently, food intake and body weight gain returned to normal (although body weight remained elevated) up to 6 mo. By this time several fat depots were still reduced in size, although total (chemical) fat was identical in treated and control groups. These results suggest that 1) reduction of body fat depots can be achieved using antibodies to adipocytes, 2) compensatory increases in lean body mass can occur, and 3) total fat mass may be regulated.
Endothelin increases the synthesis and secretion of atrial natriuretic peptide in neonatal rat cardiocytes
Authors: Gardner D. G. , Newman E. D. , Nakamura K. K. , Nguyen K. P.
Published: December 2017

Endothelin (ET) effected a dose-dependent increment in atrial natriuretic peptide (ANP) secretion and ANP mRNA accumulation in neonatal rat atrial and ventricular cardiocytes but had no effect on the processing of the ANP prohormone to the mature ANP product. The secretagogue effect was not limited by cell density. Both basal and ET-dependent secretory activity were abrogated by the calmodulin antagonist calmidazolium but were unaffected by meclophenamate or pertussis toxin. The magnitude of the ET-dependent increment in ANP secretion was amplified by culturing the cells in a dynamically pulsating (vs. static) environment, implying an interaction between mechanical and agonist-mediated secretory stimuli in this system. ET also promoted immunoreactive ANP release from primary cultures of fetal rat hypothalamic cultures, suggesting that this regulatory function may be generally employed in ANP gene-expressing cells. These findings demonstrate that ET has parallel effects on ANP synthesis and secretion and support a role for this peptide in the regulation of local and circulating levels of the natriuretic hormone.
Postnatal changes in lambs of two pathways for thyroxine 5'-monodeiodination in brown adipose tissue
Authors: Wu S. Y. , Kim J. K. , Chopra I. J. , Murata Y. , Fisher D. A.
Published: December 2017

We have recently shown that ovine fetal brown adipose tissue (BAT) contains two distinct iodothyronine 5'-monodeiodinase (5'MDI) activities, one with a high Km (type I) and another with a low Km (type II). Both activities increased to maximum levels near term (150 days gestation). BAT plays a major role in neonatal temperature regulation in lambs, and available evidence suggests that BAT 5'MDI activity is closely linked to thermogenic capacity. To better characterize the changes in 5'MDI after birth, we studied both type I and type II 5'MDI in lamb BAT from the time of birth to 30 days of postnatal age. Type I 5'MDI activity [pmol 3,5,3'-triiodothyronine (T3).mg protein-1.h-1] showed no significant changes during the first 11 days after birth [newborn (NB), 95 +/- 16; 1 day, 83 +/- 20; 3-4 days, 80 +/- 11; 10-11 days, 92 +/- 28]. Activity decreased significantly at 30 days (24 +/- 8.9, P less than 0.05). On the other hand, the type II 5'MDI activity (fmol I- released.mg protein-1.h-1) increased significantly (P less than 0.01) during the first 4 days, (NB, 348 +/- 23; 1 day, 679 +/- 37; 3-4 days, 785 +/- 199), decreased toward NB values (401 +/- 87) at 10-11 days of age, and fell to 66 +/- 31 at 30 days (P less than 0.05 vs. NB). Kinetic analysis of BAT type II thyroxine 5'MDI revealed a rise in maximum velocity from NB to 1 and 3-4 days of age without a change in the enzymatic activity Km.(ABSTRACT TRUNCATED AT 250 WORDS)
Homologous amino-terminal radioimmunoassay for rat parathyroid hormone
Authors: Calvo M. S. , Gundberg C. M. , Heath H. , Fox J.
Published: December 2017

Existing radioimmunoassays for parathyroid hormone (PTH) in rat plasma are based on cross-reactivity of rat PTH (rPTH) with heterologous antisera. We used the synthetic NH2-terminal fragment of rPTH [rPTH-(1-34)] to develop a homologous radioimmunoassay for circulating PTH. An antiserum to rPTH-(1-34) was raised in a goat (G-813), and the same peptide was used as radioligand (125I) and standard. Purification of the label by high-performance liquid chromatography (HPLC) increased specific binding greater than twofold and sensitivity by 50-100%. With a final antiserum dilution of 1:70,000, maximum specific binding of 30-33%, nonspecific binding of 1-5%, and 50-microliters sample additions, the assay detection limit was 1.8-2.5 pmol/l. A midregional fragment of human PTH did not displace 125I-labeled rPTH-(1-34). HPLC of extracts of rat parathyroid glands and hyperparathyroid plasma showed only a single peak of immunoreactivity that eluted 2 min after rPTH-(1-34). Dose dilution curves for rat parathyroid gland extracts, rPTH-(1-34) added to rat plasma, and endogenous rat plasma PTH all paralleled the standard curve. Immunoreactive PTH (irPTH) was detectable in greater than 90% of fasting normal rat plasma and changed appropriately in response to hyper- and hypocalcemia induced by low-calcium and vitamin D-deficient diets, injections of calcium and EDTA, and after thyroparathyroidectomy. The normal range for rat plasma irPTH was less than 2.0-12 pmol/l, in general agreement with bioassay results of others. Thus rPTH-(1-34) is an excellent immunogen for raising antisera to rPTH, and assays incorporating it may be of great value in studying rat parathyroid physiology.
Arterial catecholamine responses during exercise with acute and chronic high-altitude exposure
Authors: Mazzeo R. S. , Bender P. R. , Brooks G. A. , Butterfield G. E. , Groves B. M. , Sutton J. R. , Wolfel E. E. , Reeves J. T.
Published: December 2017

Exercise at high altitude is a stress that activates the sympathoadrenal systems, which could affect responses to acute altitude exposure and promote adaptations during chronic altitude exposure. However, catecholamine levels are not clearly described over time at high altitude. In seven male volunteers (23 yr, 72 kg), resting arterial norepinephrine concentrations (ng/ml) on arrival at Pikes Peak (0.338 +/- 0.041) decreased compared with sea-level values (0.525 +/- 0.034) but increased to above sea-level values after 21 days at 4,300 m (0.798 +/- 0.052). Furthermore, during 45 min of constant submaximal exercise, values were similar at sea level (1.670 +/- 0.221) and on acute exposure to 4,300 m (2.123 +/- 0.086) but increased after 21 days of chronic exposure (2.693 +/- 0.216). By contrast, resting arterial epinephrine values (ng/ml) during acute and chronic exposure (0.708 +/- 0.033 vs. 0.448 +/- 0.026) both exceeded those of sea level (0.356 +/- 0.020). During exercise values on arrival were greater than at sea level (0.921 +/- 0.024 vs. 0.397 +/- 0.035) but fell to 0.612 +/- 0.025 ng/ml after 21 days. Exercise norepinephrine levels were related to systemic vascular resistance measurements (r = 0.93), whereas epinephrine levels were related to circulating lactate (r = 0.95). We conclude that during exercise at altitude there is a dissociation between norepinephrine, an indicator of sympathetic neural activity, and epinephrine, an indicator of adrenal medullary response. These actions may account for different metabolic and physiological responses to acute vs. chronic altitude exposure.
Use of mass isotopomer distributions in secreted lipids to sample lipogenic acetyl-CoA pool in vivo in humans
Authors: Hellerstein M. K. , Kletke C. , Kaempfer S. , Wu K. , Shackleton C. H.
Published: December 2017

Measurement of hepatic fatty acid (FA) and cholesterol synthesis has been limited by lack of access to the precursor pool, cytosolic acetyl-CoA. We present a method for inferring the enrichment of the true hepatic lipogenic precursor pool in humans using the frequency distribution of mass isotopomers within enriched circulating polymers of acetyl-CoA [very low-density lipoprotein (VLDL)-palmitate, VLDL-stearate]. Human subjects were infused intravenously (n = 16) with [1-13C]- or [2-13C]acetate. Oral sulfamethoxazole (SMX) was administered concurrently, and the acetylated conjugate (SMX acetate) was used to estimate independently the hepatic cytosolic acetyl-CoA enrichment. Isotopomer frequencies in VLDL-FA were determined by gas chromatography-mass spectrometry, whereas high-performance liquid chromatography-mass spectrometry was used to measure enrichments in SMX acetate. Based on the excess M2/excess M1 ratio in VLDL-FA, calculated acetyl-CoA enrichments were 5.59 +/- 0.33 molar percent excess (MPE), whereas SMX acetate enrichments were 5.38 +/- 0.31 MPE (the 2 methods were not significantly different). Mass isotopomer-calculated and SMX acetate-measured estimates of acetyl-CoA enrichments correlated very closely in individual subjects (r2 = 0.93; P less than 0.0001). De novo hepatic lipogenesis can be measured using isotopomer-calculated precursor enrichments compared with measured incorporation in specific isotopomers of VLDL-FA. In summary, excess isotopomer frequencies in secreted lipids provide a non-invasive technique for estimating hepatic cytosolic acetyl-CoA enrichments in humans in vivo and correlate closely with enrichments observed using the xenobiotic probe technique. Isotopomeric distributions represent a new strategy for accurate measurement of macromolecule synthesis that may be applicable to other classes of molecules besides lipids.
Effect of short-term fasting on the lipolytic response to theophylline
Authors: Peters E. J. , Klein S. , Wolfe R. R.
Published: December 2017

We investigated the hypothesis that the increase in lipolysis that occurs in short-term (86-h) fasting is due to a decreased inhibitory influence of adenosine. In normal volunteers who fasted for 14 and 86 h, the response to adenosine receptor blockade was assessed by the infusion of theophylline at a rate sufficient to produce plasma concentrations (30 microM) that blocked adenosine receptors but that were well below the threshold for inhibition of phosphodiesterase. Lipolysis was assessed by determining the rate of appearance of glycerol using D-5-glycerol infusion. Fatty acid flux was also determined by means of [1-13C]palmitate infusion, and total fatty acid oxidation was determined by indirect calorimetry. There was a mild stimulatory effect of theophylline on lipolysis at 14 h. After the subjects fasted for 86 h, theophylline infusion caused a much greater increase in both lipolysis and fatty acid oxidation. These results suggest that the inhibitory effect of adenosine on lipolysis is increased during short-term fasting.
Hypertension without peripheral insulin resistance in spontaneously hypertensive rats
Authors: Buchanan T. A. , Sipos G. F. , Madrilejo N. , Liu C. , Campese V. M.
Published: December 2017

We performed euglycemic clamp studies with labeled glucose to measure insulin's effect on hepatic glucose output (HGO) and peripheral glucose clearance in eight conscious mobile spontaneously hypertensive rats (SHR) and eleven normotensive Wistar-Kyoto (WKY) rats age 9-10 wk. Systolic blood pressure was elevated in the SHR (P less than 0.001), whereas means of 12-h-fasted plasma insulin (P greater than 0.4), glucose (P greater than 0.07), HGO (P greater than 0.25), and glucose clearance (P greater than 0.2) did not differ significantly between groups. Infusions of human insulin into SHR and WKY rats (1 and 1.5 mU.min-1.kg-1, respectively) during concomitant somatostatin administration reestablished basal insulinemia in both groups. Neither HGO (P greater than 0.15) nor glucose clearance (P greater than 0.3) differed significantly between SHR and WKY rats under those conditions. Somatostatin plus higher-dose insulin infusions (4 mU.min-1.kg-1 in SHR and 3 or 6 mU.min-1.kg-1 in WKY rats) resulted in physiological hyperinsulinemia in all rats. Insulin sensitivity, calculated as the increase in glucose clearance effected by an increase in circulating insulin during higher-dose insulin infusions, did not differ significantly between SHR and WKY groups (P greater than 0.3). HGO was completely suppressed in SHR and WKY rats during the higher-dose insulin infusions. Our data indicate that hypertension is present in SHR at an age when insulin-mediated glucose disposal is not different from age-matched WKY rats. These findings do not support a role for peripheral insulin resistance in the genesis of hypertension in SHR.
Oxidation of leucine and alpha-ketoisocaproate to beta-hydroxy-beta-methylbutyrate in vivo
Authors: Van Koevering M. , Nissen S.
Published: December 2017

The oxidation of alpha-ketoisocaproate (KIC) to beta-hydroxy-beta-methylbutyrate (HMB) by the enzyme KIC dioxygenase has been previously described in rat and human liver; however, the importance of this pathway in normal leucine metabolism has not yet been assessed. A series of experiments was conducted in young lambs and pigs to determine whether HMB is produced from KIC in vivo and to estimate the importance of this pathway in leucine metabolism. In the first study, lambs were fed a bolus of KIC, and the change in plasma HMB concentration was monitored over a 24-h period. Administration of KIC increased plasma HMB from basal concentrations of 2 to approximately 7 microM 4 h after the supplementation. In the second experiment, lambs were infused with [6,6,6-2H3]KIC into the duodenum, and the appearance of labeled [2H3]HMB was measured. Under basal conditions, a minimum of 18% of the HMB was derived from KIC, but when unlabeled KIC was infused into the duodenum at a rate of 1.6 mumol.kg-1.min-1, plasma HMB concentration doubled, and essentially 100% of the HMB present was derived from KIC. In a third experiment, young pigs were infused with [6,6,6-2H3]leucine. At steady state, [2H3]-leucine and HMB enrichments were nearly identical, indicating that plasma HMB is derived solely from leucine. In a fourth experiment, both lambs and pigs were injected intravenously with 600 mg of HMB daily, and urinary HMB excretion was quantitated.(ABSTRACT TRUNCATED AT 250 WORDS)
Termination of the peripubertal FSH increase in male rats
Authors: Nazian S. J. , Cameron D. F.
Published: December 2017

To determine if the pubertal testosterone rise plays a role in the termination of the peripubertal follicle-stimulating hormone (FSH) increase, male rats were injected with ethylene dimethanesulfonate (EDS) at 40 days of age to eradicate the Leydig cells just before the onset of the testosterone rise. Rats were decapitated at weekly intervals from age 26 to 96 days. Compared with vehicle-injected controls, EDS treatment resulted in a delay in the peripubertal increase in the relative weights of prostates and seminal vesicles of approximately 2 wk. Serum testosterone remained at pretreatment levels for 1 wk postinjection. Testicular interstitial fluid testosterone remained at pretreatment concentrations for considerably longer and was significantly lower than controls for 2 wk postinjection. EDS treatment resulted in serum FSH levels that were elevated by 1 wk postinjection. They remained significantly higher than controls until 96 days of age. Compared with controls, serum alpha-inhibin was elevated after EDS as was serum luteinizing hormone. These results suggest that the pubertal testosterone increase plays an important role in terminating the peripubertal FSH rise.
Effect of hypercorticism on regulation of skeletal muscle glycogen metabolism by epinephrine
Authors: Coderre L. , Srivastava A. K. , Chiasson J. L.
Published: December 2017

The effect of hypercorticism on the regulation of glycogen metabolism by epinephrine was examined in skeletal muscles using a hindlimb perfusion technique. Rats were injected with either saline or dexamethasone (0.4 mg.kg-1.day-1) for 14 days and were studied in the fed and fasted (24 h) states under saline or epinephrine (10(-7) M) treatment. In the fed state, dexamethasone administration did not affect basal glycogen concentration but decreased glycogen synthase activity ratio in white and red gastrocnemius muscles. Epinephrine failed to decrease glycogen content despite the expected activation of glycogen phosphorylase in the fed dexamethasone-treated rats. Dexamethasone treatment resulted in a threefold increase in the level of muscle adenosine, a phosphorylase a inhibitor. In control rats, fasting was associated with a decrease in muscle glycogen concentration (P less than 0.01) and with an increase in the glycogen synthase activity ratio. Dexamethasone treatment, however, totally abolished both the decreased muscle glycogen content and glycogen synthase activation observed in fasting controls. In the dexamethasone-treated group, fasting restored the glycogenolytic effect of epinephrine. Interestingly, it was associated with decreased muscle adenosine concentrations. These data indicate that, in the fed state, dexamethasone treatment inhibits skeletal muscle glycogenolysis in response to epinephrine despite phosphorylase activation and glycogen synthase inactivation. It is suggested that this abnormality could be due to the inhibition of phosphorylase a by increased muscle adenosine levels.
Leucine and phenylalanine kinetics during mixed meal ingestion: a multiple tracer approach
Authors: Biolo G. , Tessari P. , Inchiostro S. , Bruttomesso D. , Fongher C. , Sabadin L. , Fratton M. G. , Valerio A. , Tiengo A.
Published: December 2017

To estimate whole body and splanchnic metabolism of dietary amino acids, phenylalanine and leucine kinetics were determined simultaneously in six normal volunteers before and during the constant administration of a complete mixed meal, employing multiple tracers of these amino acids. L-[5,5,5-2H]leucine and L-[2,6-3H]-phenylalanine were infused intravenously; L-[1-13C]leucine and L-[1-14C]phenylalanine were administered orally with the meal. During the meal, steady-state leucine concentration rose from 136 +/- 6 to 190 +/- 14 mumol/l (P less than 0.01), phenylalanine from 44 +/- 4 to 61 +/- 6 mumol/l (P less than 0.01), total leucine rate of appearance (Ra) from 1.29 +/- 0.03 to 1.77 +/- 0.07 (P less than 0.01, +37 +/- 3%), and phenylalanine Ra from 0.73 +/- 0.05 to 0.80 +/- 0.07 mumol.kg-1.min-1 (P less than 0.05, +8 +/- 3%). Splanchnic uptake of dietary phenylalanine was greater (P less than 0.001) than that of leucine (58 +/- 4 vs. 25 +/- 4%, respectively), 44 +/- 3% of circulating leucine derived from the diet vs. 20 +/- 2% of circulating phenylalanine (P less than 0.01). Endogenous leucine and phenylalanine Ra were significantly suppressed (P less than 0.05). In summary: 1) splanchnic uptake of dietary phenylalanine is onefold greater than that of leucine; 2) dietary contribution to systemic phenylalanine Ra is about half of that to leucine Ra; and 3) endogenous appearance of both leucine and phenylalanine after the meal is suppressed. In conclusion, splanchnic metabolism of dietary leucine and phenylalanine differs markedly and can be quantitated in vivo without catheterization.