bluuebunny/crossref_metadata_2025 huggingface dataset:
Altered hepatic mitochondrial fatty acid oxidation and ketogenesis in endotoxic rats
Authors: Takeyama N. , Itoh Y. , Kitazawa Y. , Tanaka T.
Published: December 2017

Rat hepatic mitochondrial function, including oxidative phosphorylation, fatty acid oxidative capacity, kinetic parameters of carnitine palmitoyltransferase I (CPT I), and sensitivity of CPT I to malonyl-CoA inhibition were studied in vitro in isolated mitochondria following Escherichia coli lipopolysaccharide (LPS). The hepatic mitochondrial CPT I in LPS-treated rats showed a lower apparent maximum velocity (Vmax) for palmitoyl-CoA and Ki for malonyl-CoA without changes in apparent Km for palmitoyl-CoA. The rate of oxygen consumption or end-product formation of palmitoyl-L-carnitine and octanoate was not altered, but the rate of CPT I-dependent palmitoyl-CoA (plus L-carnitine) oxidation was reduced by LPS, when acetyl-CoA produced via beta-oxidation was directed toward citrate. When acetyl-CoA was directed to acetoacetate, the oxygen consumption rates of palmitoyl-L-carnitine and palmitoyl-CoA (plus L-carnitine) were decreased by LPS, although mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase activity was not altered. These results indicate that hepatic mitochondria isolated from LPS-treated rats show lower ketogenic and long-chain acyl-CoA oxidative capacity than those of fasted controls, and inhibition of ketogenesis is elicited at a site distal to CPT I in addition to reduction in CPT I activity.
Formation of gluconeogenic precursors in rat skeletal muscle during fasted-refed transition
Authors: Goodman M. N. , Dietrich R. , Luu P.
Published: December 2017

During the fasted-refed transition, hepatic glycogen repletion from glucose can occur by the direct and indirect pathway. In the indirect pathway, glucose is first metabolized to 3-carbon intermediates that then are converted in the liver to glucose 6-phosphate via the gluconeogenic pathway before conversion to glycogen. The present study evaluated whether skeletal muscle is a major source of 3-carbon intermediates (i.e., lactate, pyruvate, and alanine) during refeeding of 1-day fasted rats. Arteriovenous differences for lactate, pyruvate, and alanine across the anesthetized rat hindlimbs were used to evaluate muscle metabolism in the fed, fasted, and refed state. In the fasted state, liver glycogen was depleted, and muscle released 3-carbon intermediates. One hour after refeeding, hepatic glycogen was 30% repleted, and blood lactate, pyruvate, and alanine increased. Despite this, the release of alanine by muscle diminished at this time and lactate was removed. At 4 h after refeeding, 3-carbon intermediates were all released by hindlimb tissue but in an amount not greater than in the fasted state. Overall, these results suggest that skeletal muscle in the rat is not a major source of 3-carbon precursors for early postprandial hepatic glycogen repletion via the indirect pathway, nor is the rise in 3-carbon intermediates in blood during refeeding caused by their increased output by muscle.
Effect of endurance training on glucose transport capacity and glucose transporter expression in rat skeletal muscle
Authors: Ploug T. , Stallknecht B. M. , Pedersen O. , Kahn B. B. , Ohkuwa T. , Vinten J. , Galbo H.
Published: December 2017

The effect of 10 wk endurance swim training on 3-O-methylglucose (3-MG) uptake (at 40 mM 3-MG) in skeletal muscle was studied in the perfused rat hindquarter. Training resulted in an increase of approximately 33% for maximum insulin-stimulated 3-MG transport in fast-twitch red fibers and an increase of approximately 33% for contraction-stimulated transport in slow-twitch red fibers compared with nonexercised sedentary muscle. A fully additive effect of insulin and contractions was observed both in trained and untrained muscle. Compared with transport in control rats subjected to an almost exhaustive single exercise session the day before experiment both maximum insulin- and contraction-stimulated transport rates were increased in all muscle types in trained rats. Accordingly, the increased glucose transport capacity in trained muscle was not due to a residual effect of the last training session. Half-times for reversal of contraction-induced glucose transport were similar in trained and untrained muscles. The concentrations of mRNA for GLUT-1 (the erythrocyte-brain-Hep G2 glucose transporter) and GLUT-4 (the adipocyte-muscle glucose transporter) were increased approximately twofold by training in fast-twitch red muscle fibers. In parallel to this, Western blot demonstrated a approximately 47% increase in GLUT-1 protein and a approximately 31% increase in GLUT-4 protein. This indicates that the increases in maximum velocity for 3-MG transport in trained muscle is due to an increased number of glucose transporters.
Ca2+ release and InsP3 production in avian uterine cells: effects of PGF2 alpha and AVT
Authors: Molnar M. , Hertelendy F.
Published: December 2017

Primary cultures of smooth muscle cells isolated from the shell gland ("uterus") of the domestic hen were permeabilized with digitonin and loaded with 45Ca2+ in the presence of ATP. When these cells were stimulated with prostaglandin F2 alpha (PGF2 alpha), arginine vasotocin (AVT), or D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3], there was a rapid, biphasic, and dose-related release of 45Ca2+ from nonmitochondrial pools. 2-Nitro-4-carboxyphenyl-N,N-diphenylcarbamate, an inhibitor of phospholipase C, had no effect on PGF2 alpha - and Ins(1,4,5)P3-promoted 45Ca2+ efflux, whereas it significantly inhibited AVT-stimulated and a stable analogue of GTP-stimulated 45Ca2+ release. In fura-2-loaded intact cells, both PGF2 alpha and AVT increased intracellular Ca2+ levels [( Ca2+]i) in a dose-related manner in the presence of extracellular Ca2+. However, omission of extracellular Ca2+ prevented a PGF2 alpha, but not AVT-induced, rise in [Ca2+]i In D-myo-[3H]inositol-labeled cells, 10 nM AVT caused a rapid, two- to threefold increase in [3H]-Insp3, whereas PGF2 alpha up to 1 microM was infective. Raising PGF2 alpha to 10 microM increased total inositol phosphates by 22% over controls (P less than 0.05). These results point to marked differences in the mechanisms by which AVT and PGF2 alpha regulate [Ca2+]i in uterine smooth muscle cells. It is suggested that the two agonists act in concert to initiate oviposition.
Distinct but nonadditive effects of epinephrine and cortisol on determinants of glucose tolerance in dogs
Authors: Martin I. K. , Christopher M. J. , Alford F. P. , Best J. D.
Published: December 2017

Effects of physiological increments of epinephrine (Epi) and cortisol (F) on glucose metabolism were assessed in dogs just before and during an intravenous glucose tolerance test performed in the last 3 h of an acute (short F + Epi, 4 h F and 3.5 h Epi) or prolonged (long F + Epi, 75 h F and Epi) infusion period. Comparison of the F + Epi effects with those of F and Epi alone enabled us to describe interactions between these hormones. The increase in plasma glucose after long F + Epi [from control (saline, Sal) of 5.2 +/- 0.1 to 5.8 +/- 0.1 mmol/l; n = 8; P less than 0.01] was not greater than the sum of the glucose increments after long F and long Epi individually. Long Epi and long F both reduced glucose tolerance (KGlc) significantly, but the decline during long F + Epi (from Sal 3.6 +/- 0.7 to 2.9 +/- 0.5%/min; P greater than 0.1) was less than during either individual infusion. Minimal model analysis showed that F attenuated the inhibitory effects of long Epi on glucose-mediated glucose disposal (SGlc), so that it was not reduced from 3.8 +/- 0.8 (Sal) during long F + Epi compared with the fall to 1.3 +/- 0.7 x 10(-2) min-1 (n = 6; P less than 0.05) during long Epi alone. F had the dominant influence on insulin sensitivity (SI) during infusion of F + Epi. The reduction of SI from 8.4 +/- 1.1 (Sal) to 6.6 +/- 1.2 (short F + Epi) and 5.1 +/- 1.1 x 10(-4) min-1 per mU/l (long F + Epi; P less than 0.05) paralleled that seen with F alone but contrasted with the acute reduction of SI during short Epi (4.8 +/- 1.5; P less than 0.02 vs. Sal) and its restoration to control values of 9.0 +/- 2.1 x 10(-4) min-1 per mU/l during long Epi. We conclude that Epi and F have distinct but nonadditive effects on determinants of glucose tolerance.
Body composition and metabolic rate in rat during a continuous infusion of cachectin
Authors: Hoshino E. , Pichard C. , Greenwood C. E. , Kuo G. C. , Cameron R. G. , Kurian R. , Kearns J. P. , Allard J. P. , Jeejeebhoy K. N.
Published: December 2017

Changes in brain metabolites, energy balance, resting metabolic rate, body composition, and organ histology were studied over 10 days in control (C), cachectin-infused (CIR), and pair-fed (PFR) (in relation to CIR) rats. The cachectin was continuously infused for the 10 days into the superior vena cava at the rate of approximately 100 micrograms.kg-1.day-1. The brain of the CIR had significantly more tryptophan and 5-hydroxyindole-3-acetic acid than C and PFR. The CIR rats were anorexic, hypermetabolic, relatively hyperglycemic, and had raised blood urea nitrogen with comparable creatinine levels when compared with similarly wasted PFR. They had significant loss of muscle mass, especially in muscles with a predominance of type II fibers. However they gained liver, heart, and lung mass. The loss of muscle mass could be ascribed to dietary deficiency, but the gain in visceral mass was associated with an increase in organ DNA and protein. Histology showed that there was cell proliferation in the liver, heart, and kidneys. The data are consistent with centrally mediated anorexia with nutritionally mediated muscle wasting but with visceral hypermetabolism, protein accumulation, and cell proliferation.
Energy equivalents of CO2 and their importance in assessing energy expenditure when using tracer techniques
Authors: Elia M.
Published: December 2017

Carbon dioxide production in free living animals and humans can be measured using tracer techniques, but the prediction of energy expenditure also requires an estimate of the energy equivalents of CO2 (energy expended/CO2 produced; EeqCO2). This work is concerned with assessing the variation in EeqCO2 with the use of dietary information, indirect calorimetry, and theoretical concepts. The EeqCO2 for diets (EeqCO2 diet) ingested by 63 individuals living in a Cambridgeshire village, UK, was found to vary by less than 10%. The EeqCO2 diet for different populations varied by greater than 10% and for artificial enteral feeds by approximately 20%. Alcohol increases this variability because it has a particularly high EeqCO2. Variation in the nitrogenous end products of metabolism may also have a substantial effect on the EeqCO2 for a subject (EeqCO2 body), especially when a large proportion of energy expenditure is derived from protein oxidation, as in strict carnivores. Nutrient/energy imbalances such as those associated with growth, hypercaloric feeding, or starvation may also have major effects on EeqCO2 body. It is concluded that the calculation of energy expenditure from CO2 production should not employ a universal value for EeqCO2 body. The value should take into account the physiological and clinical state under investigation. Practical recommendations are suggested.
Functional and metabolic consequences of skeletal muscle remodeling in hypothyroidism
Authors: McAllister R. M. , Ogilvie R. W. , Terjung R. L.
Published: December 2017

Functional and metabolic responses of hypothyroid skeletal muscle were evaluated during steady-state isometric contraction conditions, using an isolated perfused rat hindlimb preparation. Treating rats with propylthiouracil (PTU) for 4-5 mo resulted in a 55% decrease (P less than 0.001) in citrate synthase activity in plantaris muscle and phenotypic remodeling of the plantaris, evident by a threefold increase in type I fiber area and a 13% decrease in type II fiber area. Perfusion of PTU (n = 9) and control (n = 9) rat hindlimbs of similar size, with similar inflow (approximately 10 ml/min) and oxygen content (approximately 20 g/100 ml), resulted in similar oxygen deliveries to the contracting muscles (PTU 11.4 +/- 0.58, control 9.54 +/- 0.75 mumol.min-1.g-1; P greater than 0.05). Ten-minute tetanic contraction (100 ms at 100 Hz) periods at 4, 8, 15, 30, and 45 tetani/min were elicited in consecutive ascending order. Oxygen consumption (VO2) was lower in the PTU group at all contraction frequencies (P less than 0.005), with a decrease in peak VO2 of 44% (PTU 3.01 +/- 0.29, control 5.35 +/- 0.42 mumol.min-1.g-1; P less than 0.001). Oxygen extraction by the PTU muscle was only approximately 25% of that delivered. Developed tension was initially less (15%; P less than 0.05) in the PTU group but declined in a similar manner, as a percent of initial, to that of the control group. The slightly lower absolute tension development of the PTU muscle could not account for the large reduction in VO2.(ABSTRACT TRUNCATED AT 250 WORDS)
Description of a direct-indirect room-sized calorimeter
Authors: Seale J. L. , Rumpler W. V. , Moe P. W.
Published: December 2017

A calorimeter suitable for measuring human energy expenditure has been assembled by the US Department of Agriculture in Beltsville, Maryland. The room-sized calorimeter is 3.05 X 2.74 X 2.44 m (20.39 m3). Direct and indirect calorimetry methods are used to simultaneously measure heat emission and energy expenditure. A water-cooled gradient layer chamber is used to measure human heat production directly. Indirect calorimetry is ascertained by measuring the changes in gas composition of the air entering and existing the chamber. The inlet and outlet air are each sampled three times every 100 s with a multiple gas analyzer to determine carbon dioxide and methane production and oxygen consumption within the chamber. A total of 30 measurements, which include temperature, pressure, gas fractions, flow rates, direct heat transfer, electrical power, and motion within the chamber, are converted from electrical to digital signals and recorded on magnetic disk nine times each minute. Real-time calculations for directly and indirectly measured energy expenditure are continuously updated and displayed. The performance of the calorimeter is assessed by the combustion of alcohol within the chamber. Results of these tests indicate that the direct heat recovery is 99.7 +/- 2.6 (SD) % and the indirect heat recovery is 100.3 +/- 1.3 (SD) %.
Energy expenditure and substrate metabolism in ethanol-induced liver cirrhosis
Authors: Muller M. J. , Fenk A. , Lautz H. U. , Selberg O. , Canzler H. , Balks H. J. , von zur Muhlen A. , Schmidt E. , Schmidt F. W.
Published: December 2017

Energy expenditure and substrate metabolism were investigated in 10 patients with alcoholic liver cirrhosis (EtOH-Ci) and 10 healthy controls (C). Resting metabolic rate (RMR) varied from 1,269 to 2,467 kcal/day in C and from 1,228 to 2,098 kcal/day in EtOH-Ci. RMR was significantly related to fat-free mass (FFM) in both groups, but EtOH-Ci decreased FFM and increased RMR when expressed per kilogram FFM (+33%). Glucose intolerance, hyperinsulinemia, and a decreased C-peptide-to-insulin ratio were observed in EtOH-Ci after a test meal. Concomitantly, nonoxidative glucose metabolism was reduced in association with normal increases in glucose oxidation. EtOH-Ci reduced insulin sensitivity (-59%) and maximal insulin-dependent glucose disposal (-40%) during a sequential two-step glucose clamp protocol (phase 1: 1 mU.kg body wt-1.min-1 insulin infusion rate + euglycemia; phase 2: 4 mU.kg body wt-1.min-1 insulin infusion rate + 165 mg/dl plasma glucose concentration). This was explained by reduced glucose storage (-99%, -51%) in association with normal responses in glucose oxidation rate, plasma lactate concentration, lipid oxidation rate, and rate of lipogenesis. Defective glucose storage was independent of reduced FFM. EtOH-Ci increased glucose-induced thermogenesis by 57%. We conclude that increased resting metabolic rate, enhanced thermogenesis, defective glucose storage, and normal glucose oxidation together result in increased energy needs and favor negative energy balance in patients with alcoholic cirrhosis.
Dual mechanism of insulin action on human skeletal muscle: identification of an indirect component not mediated by FFA
Authors: Capaldo B. , Napoli R. , Di Bonito P. , Albano G. , Sacca L.
Published: December 2017

To determine whether insulin action on human skeletal muscle is entirely accounted for by a direct effect of insulin per se, we quantitated forearm glucose uptake (FGU) in two groups of normal subjects under conditions of identical forearm tissue insulinization (approximately 90 microU/ml) induced by means of systemic (group I) or intrabrachial insulin infusion (group II). With this approach, a difference of FGU between the two groups is demonstrative of the operation of indirect mechanisms in insulin action. During insulin infusion in group I, euglycemia was maintained by a variable glucose infusion, whereas arterial free fatty acid (FFA) concentration fell below 0.1 mM. In group II, arterial concentrations of both glucose and FFA remained unchanged, indicating that insulin effect was indeed restricted to the forearm tissues. With local insulin administration, FGU reached levels approximately 40% lower than those achieved with systemic insulin infusion (P less than 0.01-0.005). To determine whether this difference could be attributed to the fall in FFA concentration associated with systemic insulinization, FGU was also quantitated during a systemic insulin plus heparin infusion (group III) to maintain FFA at their basal levels. In this condition, insulin-stimulated FGU was not different from that observed in group I. These data demonstrate that 1) insulin action on skeletal muscle glucose disposal is mediated by a direct as well as an indirect component, and 2) the maintenance of basal FFA does not affect insulin-stimulated FGU, indicating that the indirect component of insulin action is mediated by a different mechanism.
Interrelationship between hepatic ureagenesis and gluconeogenesis in early sepsis
Authors: Ohtake Y. , Clemens M. G.
Published: December 2017

This study was performed to investigate the interrelationship between gluconeogenesis and ureagenesis during sepsis. In isolated perfused livers, gluconeogenesis was assessed using either lactate or a combination of lactate, glutamine, and alanine as substrate. Ureagenesis was assessed using either NH4Cl or glutamine plus alanine as substrate. NH4Cl stimulated urea production in livers from both septic and sham-operated control rats. Urea release was approximately 1.2 and 2.0 mg urea nitrogen.g-1.h-1 for 1 and 5 mM NH4Cl, respectively, and was equal for both groups. With amino acids as substrate, urea production was significantly greater in livers from septic animals compared with controls. Phenylephrine stimulated urea production in the sham-operated group by about twofold, whereas in the septic group urea release was slightly inhibited. Gluconeogenesis from lactate was inhibited by NH4Cl (1 and 5 mM) in both groups, with no difference between groups. In contrast to enhanced ureagenesis from amino acids in septic rats, gluconeogenesis was decreased by approximately 24% (P less than 0.5). Similarly, phenylephrine (1 microM) stimulated gluconeogenesis by 13 +/- 1 mumol.g-1.h-1 in sham-operated rats but only by 9 +/- 1 mumol.g-1.h-1 in septic rats (P less than 0.02). These results suggest that hepatic gluconeogenic and ureagenic pathways are intact in sepsis but that altered substrate preference and hormone sensitivity may result in decreased gluconeogenesis in the presence of elevated amino acid levels.
Increased insulin action in rats with mild insulin deficiency induced by neonatal streptozotocin
Authors: Kergoat M. , Guerre-Millo M. , Lauva M. , Portha B.
Published: December 2017

The effect of chronic moderate insulin deficiency on in vivo insulin action was studied in young rats after neonatal (day of birth) streptozotocin (n0-STZ model) at a time (4 wk) when basal hyperglycemia is not yet established. The insulin action was quantified in vivo during insulin-glucose clamps performed on anesthetized rats while in the postabsorptive state; under basal or hyperinsulinemic conditions, total glucose production and utilization were assessed with a [3- 3H]-glucose perfusion, and local glucose utilization was estimated by measuring 2-deoxy-D-[1-3H]glucose 6-phosphate accumulation in various tissues. Compared with controls, the 4-wk-old n0-STZ rats were normoglycemic and hypoinsulinemic (P less than 0.05). The basal glucose production rate in the diabetics was significantly greater (P less than 0.05). During the clamp studies, suppression of endogenous glucose production by submaximal (1 nmol/l) or maximal (30 nmol/l) hyperinsulinemia was similar in both groups. Overall glucose utilization in the diabetics was significantly higher in the basal state and, after submaximal or maximal hyperinsulinemia, it remained significantly greater (P less than 0.001) than the corresponding utilization in the controls. This was correlated with an increased stimulation of glucose utilization in soleus muscle, diaphragm, white adipose tissue, and brown adipose tissue. In vitro studies using inguinal adipocytes showed that the glucose oxidative pathway retained normal sensitivity to insulin in the n0-STZ rats while the glucose conversion into lipids was significantly higher at sub-maximal insulin concentration compared with the control group.(ABSTRACT TRUNCATED AT 250 WORDS)
Effects of high salt intake on dopamine production in rat kidney
Authors: Hayashi M. , Yamaji Y. , Kitajima W. , Saruta T.
Published: December 2017

To examine the origin of increased urinary dopamine excretion (UDAV) during high salt intake, we measured UDAV from the innervated (INN) or the chronically denervated (DEN) kidney in rats fed either a high-salt (HS) or low-salt (LS) diet. UDAV of DEN [3.50 +/- 0.46 ng. min-1.inulin clearance (CIN)-1] and INN (4.00 +/- 0.59 ng. min-1.CIN-1) kidneys in the HS group showed a significant increase compared with that of the respective kidney in the LS group (DEN 1.42 +/- 0.12, INN 1.44 +/- 0.09 ng.min-1.CIN-1), whereas the effect of denervation on UDAV was not significantly different between two groups. We determined aromatic L-amino acid decarboxylase (L-AADC) activity and dopamine degradation rate of microdissected proximal convoluted tubules to study the changes in dopamine metabolism. L-AADC activity in the HS group showed a significant increase compared with that in the LS group, although there was no significant change in dopamine degradation rate. We conclude that the increase in UDAV during high salt intake was mainly caused by the enhancement of extraneural dopamine production by the kidney in rats. Dopamine-producing enzyme, but not its degradation in the tubular cells, plays a role in the regulation of extraneural dopamine production.
Catecholamines in prevention of hypoglycemia during exercise in humans
Authors: Marker J. C. , Hirsch I. B. , Smith L. J. , Parvin C. A. , Holloszy J. O. , Cryer P. E.
Published: December 2017

To assess the role of catecholamines in the prevention of hypoglycemia during moderate exercise (approximately 60% peak O2 consumption for 60 min), normal humans were studied with combined alpha- and beta-adrenergic blockade and with adrenergic blockade while changes in insulin and glucagon were prevented with the islet clamp technique (somatostatin infusion with insulin and glucagon infused at fixed rates). The results were compared with those from an islet clamp alone study. In contrast to a comparison study (saline infusion), adrenergic blockade resulted in a small initial decrease in plasma glucose during exercise, from 5.0 +/- 0.2 to 4.4 +/- 0.2 mmol/l (P less than 0.01), but the level then plateaued. There was a substantial exercise-associated decrement in plasma glucose when insulin and glucagon were held constant, i.e., from 5.5 +/- 0.2 to 3.4 +/- 0.2 mmol/l (P less than 0.0001), but the level again plateaued. However, when insulin and glucagon were held constant and catecholamine actions were blocked simultaneously, progressive hypoglycemia, to 2.6 +/- 0.6 mmol/l (P less than 0.001), developed during exercise. Hypoglycemia was the result of an absent increase in glucose production and an exaggerated initial increase in glucose utilization. Thus we conclude that sympathochromaffin activation plays a minor role when insulin and glucagon are operative, but a catecholamine, probably epinephrine, becomes critical to the prevention of hypoglycemia during exercise when changes in insulin and glucagon do not occur.
Progesterone inhibits K conductance in plasma membrane of cultured renal epitheloid MDCK cells
Authors: Steidl M. , Pinggera G. , Ritter M. , Lang F.
Published: December 2017

Progesterone causes natriuresis, an effect largely attributed to displacement of aldosterone from its receptor. The present study, however, demonstrates that progesterone (0.1, 1, and 10 mumol/1, respectively) also causes a rapid, fully reversible depolarization of Madin-Darby canine kidney (MDCK) cells (by 1.3 +/- 0.5, 4.1 +/- 0.7 and 12.3 +/- 1.5 mV, respectively). 17 alpha-Hydroxyprogesterone and dihydroxytestosterone are, by two orders of magnitude, less effective, whereas cholesterol, aldosterone, hydrocortisone, and estradiol (each up to 10 mumol/l) did not significantly alter the potential difference across the cell membrane. The effect of progesterone is blunted by antiprogestogen RU 486 (5 mumol/l). The progesterone-induced depolarization is paralleled by a decrease of potassium selectivity and an increase of cell membrane resistance and is abolished in the presence of the potassium channel blocker barium (10 mmol/l), as well as in the presence of 40 mmol/l potassium in the extracellular fluid. Neither removal of extracellular chloride or bicarbonate nor amiloride, ouabain, or pretreatment with pertussis toxin abolish the depolarizing effect of 5 mumol/l progesterone. In conclusion, acute administration of progesterone depolarizes MDCK cells by decreasing the potassium conductance of the cell membrane.
Fluoride selectively stimulates Na-dependent phosphate transport in osteoblast-like cells
Authors: Selz T. , Caverzasio J. , Bonjour J. P.
Published: December 2017

The influence of fluoride (F) on the transport of Pi was investigated in the osteoblast-like cell line UMR-106. Exposure of cells to F induced a dose-related stimulation of the Na-coupled Pi transport. Pi transport was significantly increased 6 h after 1 mM F incubation, with maximal response observed at 24 h (F 38.0 +/- 2.3, vehicle 19.8 +/- 1.2 pmol.micrograms DNA-1.4 min-1; P less than 0.001). Na-dependent alanine transport was not changed by F. The selective effect of F on Pi transport was not associated with changes in adenosine 3',5'-cyclic monophosphate, cell proliferation, or alkaline phosphatase activity. However, it was completely blunted by inhibiting translational processes with cycloheximide. Furthermore, F enhanced the stimulatory effect on Pi transport of various mitogens such as fetal calf serum, insulin, and insulin-like growth factor I. In conclusion, F can selectively enhance the activity of the Pi transport system present in the plasma membrane of UMR 106 osteoblast-like cells by a mechanism that probably involves newly synthetized proteins.
Hormone secretion stimulated by ethanol-induced cell swelling in normal rat adenohypophysial cells
Authors: Sato N. , Wang X. B. , Greer M. A.
Published: December 2017

Ethanol has been reported to affect endocrine functions, but its mechanism of action is unclear. To evaluate the hypothesis that cell swelling induced by ethanol permeation through the plasmalemma triggers hormone secretion, we studied the effect of ethanol on both hormone secretion and cell volume in acutely dispersed rat adenohypophysial cells under isotonic and hypertonic conditions. Isotonic ethanol caused a prompt cell swelling and an explosive secretory burst of prolactin and thyrotropin, which were proportional to the concentration of ethanol between 10 and 120 mM. The lowest effective dose of isotonic ethanol was 10 mM, which is below the plasma levels of legal intoxication (16 mM). Removal of medium Ca2+ enhanced the isotonic ethanol-induced increases in both cell volume and secretion. Hypertonic ethanol was ineffective in these effects. These data indicate that, in normal rat adenohypophysial cells, cell swelling caused by the rapid passage of ethanol through the plasmalemma is a potent mechanism for stimulating hormone secretion and this induced secretion is negatively modulated by extracellular Ca2+.
Smooth muscle adenosine A1 receptors couple to disparate effectors by distinct G proteins in pregnant myometrium
Authors: Schiemann W. P. , Westfall D. P. , Buxton I. L.
Published: December 2017

We have previously shown that adenosine, acting at an A1 receptor, contracts the smooth muscle of virgin guinea pig uterus (M. A. Smith, I. L. O. Buxton, and D. P. Westfall. J. Pharmacol, Exp. Ther. 247: 1059–1063, 1988) and is not coupled to the expected inhibition of adenylate cyclase (M. A. Smith, J. L. Silverstein, D. P. Westfall, and I. L. O. Buxton. Cell. Signal. 1: 357–365, 1989). To probe the importance of contractile actions of adenosine in uterine smooth muscle and to further characterize the signal transduction pathway involved in A1-receptor action, we have studied the adenosine receptor and its coupling in pregnant guinea pig myometrium. Adenosine agonist and antagonist radioligands bind to saturable sites of the A1 subtype homogeneously distributed in the smooth muscle of pregnant guinea pig uterus. Agonist competition of antagonist radioligand binding in both the absence and presence of guanine nucleotide reveals high and low agonist affinity states of the receptor. Pretreatment of tissues with pertussis toxin (PTx) shifts the high-affinity sites to a lower affinity but does not affect low-affinity sites, whereas agonist competition in the presence of guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) is indistinguishable from the control, which is consistent with coupling of A1 receptors to both PTx-sensitive and PTx-insensitive GTP-binding proteins. Adenosine receptor inhibition of adenylate cyclase activity is prevented after pretreatment of the tissue with PTx, whereas increased inositol phosphate production is not. The data presented here are consistent with coupling of the A1 receptor to dual effectors in the pregnant state of the smooth muscle. The unique action of an A1 receptor to contract mammalian smooth muscle and the appearance, only in the pregnant state, of coupling to adenylate cyclase inhibition suggest a role for adenosine in parturition biology.
Thermogenesis after surgery: effect of perioperative heat conservation and epidural anesthesia
Authors: Carli F. , Webster J. , Nandi P. , MacDonald I. A. , Pearson J. , Mehta R.
Published: December 2017

Body temperature, respiratory gas exchange, and plasma catecholamines were determined before and after surgery in three groups [control (C), warmed (W), and epidural (E) who received local anesthetic at T4-S5 dermatomes during and for 24 h after surgery] of patients undergoing colonic surgery under general anesthesia. At the end of surgery, group W were nursed in an ambient temperature of 28-30 degrees C, whereas the others were at 20-23 degrees C for a period of 24 h. Core (Tc) and dorsal hand temperature decreased during surgery in both C and E (P less than 0.05) but not in W. After surgery, Tc increased similarly in C and E and by a smaller amount in W. Plasma catecholamine concentrations increased significantly in C and W but not in E (P less than 0.001), with the greatest response occurring in C. Postoperative oxygen consumption and carbon dioxide production exceeded preoperative values (P less than 0.01) in C but not in W or E. After surgery, plasma albumin fell and C-reactive protein increased similarly in all three groups. Thus body heat conservation or epidural blockade attenuates or abolishes the rise in plasma catecholamines and oxygen consumption postoperatively but does not prevent the increase in Tc or the acute phase protein response.
On the mechanism of stimulation of ureagenesis by gluconeogenic substrates: role of pyruvate carboxylase
Authors: Martin-Requero A. , Cipres G. , Rodriguez A. , Ayuso M. S. , Parrilla R.
Published: December 2017

Gluconeogenic substrates, lactate or pyruvate, or ornithine produced 100% increase of urea synthesis from NH4Cl. The combined administration of ornithine and lactate (or pyruvate) produced more than additive effects, indicating that they acted at different steps in a potentiating manner. The uptake of ornithine was enhanced by gluconeogenic substrates. This finding may explain, at least in part, the stimulating effect of these substrates on ureagenesis from NH4Cl and ornithine. The gluconeogenic substrate-induced stimulation of ureagenesis from NH4Cl was still observed under conditions of reduced flux through pyruvate carboxylase, ruling out that their action was exclusively mediated by the anaplerotic effect of this enzyme. Pyruvate was a more potent stimulator of ureagenesis than lactate and its effect less sensitive to pyruvate carboxylase inhibition. These observations indicate that a correlation exists between stimulation of ureagenesis by gluconeogenic substrates and flux through pyruvate dehydrogenase. It is concluded that gluconeogenic substrates may stimulate ureagenesis from NH4Cl by 1) increasing intracellular ornithine availability and/or 2) enhancing flux through pyruvate dehydrogenase and consequently the tricarboxylic acid cycle activity.
Thyroid hormone effects on cardiac gene expression independent of cardiac growth and protein synthesis
Authors: Ojamaa K. , Samarel A. M. , Kupfer J. M. , Hong C. , Klein I.
Published: December 2017

Prior studies have demonstrated the importance of hemodynamic loading in mediating thyroxine (T4)-induced cardiac hypertrophy. Direct cellular effects of thyroid hormone have been implicated in modulating the expression of the myosin heavy chain (MHC) genes and the slow sarcoplasmic reticulum calcium adenosine triphosphatase (SR Ca(2+)-ATPase) gene. In the present report, administration of T4 for 72 h did not stimulate growth of the hemodynamically unloaded heterotopic isograft. The synthetic rates of total cardiac proteins and MHC in the isograft remained significantly lower at 64 and 53% of the respective rates measured simultaneously in the in situ working heart. Although total left ventricle RNA content in the isograft was unchanged by T4, alpha-MHC and SR Ca(2+)-ATPase mRNA concentrations were increased 181 and 208%, respectively, and the previously observed beta-MHC expression was completely prevented. These data indicate that, although T4 requires an increased hemodynamic load to stimulate cardiac protein synthesis, it is capable of directly altering the expression of at least two myocyte-specific genes. Therefore some of the phenotypic alterations observed with thyroid hormone treatment are the result of direct effects of the hormones on specific cardiac genes and independent of changes in cardiac growth.
Stable isotope tracer analysis by GC-MS, including quantification of isotopomer effects
Authors: Rosenblatt J. , Chinkes D. , Wolfe M. , Wolfe R. R.
Published: December 2017

In metabolic tracer studies it is frequently useful to infuse tracers that are differently labeled variants of the same molecule. These tracers are known as isotopomers. Analysis of the enrichment of each isotopic analogue can be accomplished by gas chromatography-mass spectrometry (GC-MS). However, the raw GC-MS data must be corrected to give the information required. This paper addresses how to transform the raw GC-MS data, consisting of relative abundance ratios at specific ion masses, into relative molar ratios of tracer and tracee molecules. Several correction factors are necessary. First, the background must be measured and corrected for, since it is always present in the sample. Second, the abundances in the spectrum of the labeled molecule are different from those in the unlabeled molecule, and this proportionality "skew" is corrected. A third correction factor accounts for the overlapping spectra of two or more isotopomers that cannot be measured independently. The final correction removes the "double vision" effect that may appear in some spectra due to the presence of (M - H)+ species.
Comparison of lactotrope subtypes of neonatal and adult male rats: plaque assays and patch-clamp studies
Authors: Felix R. , Horta J. , Cota G.
Published: December 2017

We examined the differences in lactotrope number and function between pituitary cultures from neonatal (10-day-old) and adult male rats. Basal hormone release was measured with the reverse hemolytic plaque assay. Whole cell Ba2+ currents through Ca2+ channels were recorded from identified prolactin (PRL) secretors with the patch-clamp technique. Lactotropes were classified in two groups according to the relative amount of PRL released: small-plaque (SP) secretors accounted for 6% of all cells in both neonatal and adult pituitary cultures, whereas large-plaque (LP) secretors comprised 13% of the adult pituitary cells but were scarce in cultures from neonates. Simultaneous plaque assays for PRL and growth hormone (GH) showed that in adults as well as in neonates the number of SP and LP secretors was similar to the number of lactosomatotropes (PRL cells that also release GH) and classical lactotropes (PRL-only cells), respectively. Ba2+ current density at positive membrane potentials was markedly higher in adult LP secretors than in neonatal or adult SP lactotropes. We conclude that the appearance of LP secretors constitutes a major postnatal change within the rat lactotrope population. These cells present a large activity of high-threshold Ca2+ channels in the plasma membrane, release PRL at high basal rates, and may correspond to classical lactotropes. The results further suggest that neonatal lactotrope-like cells persist during development and give place to adult SP secretors.
Gender differences in endocrine responses to posture and 7 days of -6 degrees head-down bed rest
Authors: Vernikos J. , Dallman M. F. , Keil L. C. , O'Hara D. , Convertino V. A.
Published: December 2017

Endocrine regulation of fluids and electrolytes during 7 days of -6 degrees head-down bed rest (HDBR) was compared in male (n = 8) and, for the first time, female (n = 8) volunteers. The subjects' responses to quiet standing for 2 h before and after HDBR were also tested. In both sexes, diuresis and natriuresis were evident during the first 2-3 days of HDBR, resulting in a marked increase in the urinary Na(+)-to-K+ ratio and significant Na+ retention on re-ambulation. After the 1st day of HDBR, plasma renin activity (PRA) was increased relative to aldosterone (Aldo), plasma volume was decreased, and the renal response to Aldo appeared to be appropriate. Circulating levels of arginine vasopressin, cortisol, and ACTH were unchanged during HDBR. Plasma testosterone decreased slightly on day 2 of HDBR in males. The ratio of early morning ACTH to cortisol was lower in females than in males because ACTH was lower in females. Urinary cortisol increased and remained elevated throughout the HDBR in males only. There were no gender differences in the responses to 7 days of HDBR, except those in the pituitary-adrenal system; those differences appeared unrelated to the postural change. The provocative cardiovascular test of quiet standing before and after HDBR revealed both sex differences and effects of HDBR. There were significant sex differences in cardiovascular responses to standing before and after HDBR. Females had greater PRA and Aldo responses to standing before HDBR and larger Aldo responses to standing after HDBR than males.(ABSTRACT TRUNCATED AT 250 WORDS)
Mechanisms of ET-1 potentiation of angiotensin II stimulation of aldosterone production
Authors: Cozza E. N. , Gomez-Sanchez C. E.
Published: December 2017

Endothelin-1 (ET-1) exerts the following two types of aldosterone-stimulating actions on glomerulosa cells: ET-1-mediated direct stimulation of aldosterone secretion (per se effect) and potentiation of the aldosterone secretion to angiotensin II (ANG II; potentiation effect). The role of Ca2+ and protein kinase C (PKC) systems in these two effects was investigated. Incubations of calf cultured adrenal zona glomerulosa cells in low-Ca2+ media or in the presence of the Ca2+ channel antagonist verapamil reduced the aldosterone secretion to ET-1. When cells were preincubated with ET-1 in a low-Ca2+ media or in the presence of the Ca2+ channel antagonist verapamil, washed, and incubated in media with normal Ca2+, ANG II showed potentiation of ANG II-stimulated aldosterone secretion. The PKC inhibitors H-7 and staurosporine did not decrease ET-1-stimulated aldosterone secretion, but they inhibited the potentiation effect of ET-1 on ANG II-mediated aldosterone secretion. Adrenocorticotropic hormone desensitization or prolonged phorbol ester stimulation of PKC resulting in desensitization also resulted in the abolition of the ET-1-mediated ANG II potentiation of aldosterone secretion. The PKC inhibitors did not affect ANG II-stimulated aldosterone secretion. We conclude that ET-1 exerts a direct stimulation of aldosterone secretion through a mechanism dependent on Ca2+ and potentiates ANG II-mediated aldosterone stimulation through a mechanism involving PKC.
Intravenous glucose tolerance test-derived glucose effectiveness in physically trained humans
Authors: Tokuyama K. , Higaki Y. , Fujitani J. , Kiyonaga A. , Tanaka H. , Shindo M. , Fukushima M. , Nakai Y. , Imura H. , Nagata I. , et al.
Published: December 2017

Glucose effectiveness (SG) and insulin sensitivity of sedentary and physically trained males were estimated by the minimal model approach. Trained subjects, who ran 86 +/- 10 km/wk and had 37% higher maximal oxygen consumption than that of sedentary subjects (56.2 +/- 1.2 vs. 40.9 +/- 1.4 ml.kg-1 x min-1, P < 0.01), were studied 16 h and 1 wk after their last training session. After overnight fasting, glucose was administered intravenously (300 mg/kg body wt) within 2 min, and insulin was infused (approximately 13-20 mU/kg given over 5 min) from 20 to 25 min after administration of glucose. Glucose disappearance constant values as an estimate of glucose tolerance were significantly higher in trained subjects after 16 h and 1 wk of their training session (3.29 +/- 0.48 and 3.60 +/- 0.64%/min) than in sedentary subjects (1.92 +/- 0.30%/min, P < 0.05). Insulin sensitivity in trained subjects measured after 16 h and 1 wk of their last training session (26.2 +/- 4.4 and 24.3 +/- 6.0 x 10(-5) min-1 x pM-1) was also higher than that of sedentary subjects (10.3 +/- 1.2 x 10(-5) min-1 x pM-1, P < 0.05). SG, the ability of glucose itself to increase peripheral glucose uptake and suppress hepatic glucose output, was significantly higher in trained subjects after 16 h and 1 wk of their last training session (0.028 +/- 0.003 and 0.030 +/- 0.004/min) than in sedentary subjects (0.017 +/- 0.002/min, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
Regulation of glucose transporter and hexokinase II expression in tissues of diabetic rats
Authors: Burcelin R. , Printz R. L. , Kande J. , Assan R. , Granner D. K. , Girard J.
Published: December 2017

Glucose transport and phosphorylation are decreased in muscle and adipose tissue in diabetes mellitus. The glucose transporter GLUT-4 and hexokinase II (HK II) are the main isoforms of proteins involved in glucose transport and phosphorylation in insulin-sensitive tissues, adipose tissue, skeletal muscle, and heart. The molecular mechanisms responsible for the decrease of glucose transport and phosphorylation have been studied during the first 3 days after streptozotocin (STZ) administration in adult male Wistar rats. GLUT-4 mRNA and protein and HK II mRNA and enzyme activity were measured. After the injection of STZ (30 h), GLUT-4 and HK II mRNAs were decreased to 10 +/- 1 and 20 +/- 3% that found in nondiabetic rats, respectively; they remained at these low levels for 72 h. Normalization of the blood glucose level by phlorizin infusion did not restore GLUT-4 and HK II mRNA concentrations to normal. In contrast, normalization of the blood glucose level by physiological infusion of insulin resulted in a total normalization of GLUT-4 and HK II mRNA concentrations. When insulin therapy was stopped, GLUT-4 and HK II mRNA and protein concentrations fell in 6 h to 40 and 20% of control levels, respectively. Minimal changes of GLUT-4 and HK II mRNA, and of HK II activity, were observed in skeletal muscle and heart of diabetic rats. We conclude that GLUT-4 and HK II mRNA are coordinately expressed in white adipose tissue. They are rapidly affected by an acute decrease of the plasma insulin concentrations but are not modified by hyperglycemia. In contrast, skeletal muscle and heart GLUT-4 and HK II mRNA are not greatly affected by short-term diabetes.
Angiotensin II is the mediator of the increase in hepatic angiotensinogen synthesis after bilateral nephrectomy
Authors: Hilgenfeldt U. , Schwind S.
Published: December 2017

The present study investigated the hypothesis that the increase in plasma angiotensinogen after nephrectomy is mediated by endogenous renin and angiotensin (ANG) II. Rats were divided into control, nephrectomy, or nephrectomy plus adrenalectomy groups. In addition, similar cohorts were divided as just mentioned and then given either atenolol (selective beta 1-adrenoceptor inhibitor that prevents renin release) or Dup-753 [ANG II (AT1) receptor antagonist]. The plasma angiotensinogen levels increase approximately fivefold after 24 h in nephrectomized rats. Pretreatment with atenolol blunted this increase. A significant reduction was observed 4 h (P < 0.05) and 8 h (P < 0.005) after surgery. Dup-753 nearly abolished the increase in angiotensinogen plasma levels. After pretreatment with Dup-753, significantly higher angiotensinogen levels (P < 0.005) were found only after 24 h. Nephrectomy plus adrenalectomy also blunted the rise in plasma angiotensinogen. A significant increase in angiotensinogen plasma levels could only be observed after 8 h (P < 0.005) and 12 h (P < 0.005) but not after 24 h. Atenolol further reduced this increase. After atenolol pretreatment, significantly higher angiotensinogen levels could only be observed after 12 h (P < 0.05). Dup-753 completely abolished the increase of plasma angiotensinogen after nephrectomy plus adrenalectomy. In anesthetized control rats at time 0 the plasma ANG I levels were 0.7 nM. Pretreatment with atenolol decreased the ANG I values by 30%, whereas Dup-753 caused a sixfold increase in plasma ANG I levels in the control rats at time 0.(ABSTRACT TRUNCATED AT 250 WORDS)