bluuebunny/crossref_metadata_2025 huggingface dataset:
Marked reduction of acyl-CoA synthetase activity and mRNA in intra-abdominal visceral fat by physical exercise
Authors: Shimomura I. , Tokunaga K. , Kotani K. , Keno Y. , Yanase-Fujiwara M. , Kanosue K. , Jiao S. , Funahashi T. , Kobatake T. , Yamamoto T. , et al.
Published: December 2017

Several reports have suggested that the reduction of intra-abdominal visceral fat after physical exercise is more prominent than that of subcutaneous fat. We compared some parameters in mesenteric and subcutaneous fats between sedentary and exercised rats (treadmill running; 10–20 m/min, 60 min/day, 7 days). Tissue weight and cell volume were decreased in mesenteric fat by the exercise. The exercise reduced activity and mRNA levels of acyl-CoA synthetase (ACS; 67 and 26% of those of the sedentary group, respectively), mRNA levels of lipoprotein lipase (LPL; 49% of those of the sedentary group), and GLUT-4 (38% of those of the sedentary group) in the mesenteric fat. In contrast, all of these parameters did not change significantly in the subcutaneous fat. Gastrocnemius muscle was heavier in exercised rats. ACS activity was elevated in the gastrocnemius muscle of the exercised rats (137% of those of sedentary group), although mRNA levels of ACS, LPL, and GLUT-4 did not change in the muscle by the exercise. These observations suggest that mesenteric fat may contribute to switching of distribution of plasma energy flux, including lipid and glucose, from fat tissue to muscle in physical exercise.
Correction of acidosis in humans with CRF decreases protein degradation and amino acid oxidation
Authors: Reaich D. , Channon S. M. , Scrimgeour C. M. , Daley S. E. , Wilkinson R. , Goodship T. H.
Published: December 2017

The effect of correction of acidosis in chronic renal failure (CRF) was determined from the kinetics of infused L-[1-13C]leucine. Nine CRF patients were studied before (acid) and after two 4-wk treatment periods of sodium bicarbonate (NaHCO3) and sodium chloride (NaCl) (pH: acid 7.31 +/- 0.01, NaHCO3 7.38 +/- 0.01, NaCl 7.30 +/- 0.01). Leucine appearance from body protein (PD), leucine disappearance into body protein (PS) and leucine oxidation (O) decreased significantly with correction of acidosis (PD: acid 122.4 +/- 6.1, NaHCO3 88.3 +/- 6.9, NaCl 116.2 +/- 9.1 mumol.kg-1.h-1, acid vs. NaHCO3 P < 0.01, NaHCO3 vs. NaCl P < 0.01, acid vs. NaCl NS; PS: acid 109.4 +/- 5.6, NaHCO3 79.0 +/- 6.3, NaCl 101.3 +/- 7.7 mumol.kg-1.h-1, acid vs. NaHCO3 P < 0.01, NaHCO3 vs. NaCl P < 0.01, acid vs. NaCl NS; O: acid 13.0 +/- 1.2, NaHCO3 9.2 +/- 0.9, NaCl 15.0 +/- 1.9 mumol.kg-1.h-1, acid vs. NaHCO3 P < 0.05, NaHCO3 vs. NaCl P < 0.01, acid vs. NaCl NS). There were no significant changes in plasma amino acid concentrations. These results confirm that correction of acidosis in chronic renal failure removes a potential catabolic factor.
Impaired febrile responses of aging mice are mediated by endogenous lipocortin-1 (annexin-1)
Authors: Strijbos P. J. , Horan M. A. , Carey F. , Rothwell N. J.
Published: December 2017

The mechanisms underlying age-related impairments in febrile responses were investigated in female C57Bl/lcrf-a(t) mice. Injection of norepinephrine, to assess total thermogenic capacity, significantly increased oxygen consumption (VO2) in all age groups, although the responses of the aged mice were significantly reduced. Injection of lipopolysaccharide or murine interleukin-1 beta (mIL-1 beta) significantly increased body temperature and VO2 in the young and adult mice but not in the aged mice. The impaired responses to mIL-1 beta in the aged mice were normalized by either injection of the glucocorticoid receptor antagonist RU-38486 or by injection of an antiserum to lipocortin-1 or its purified immunoglobulin G fraction. Injection of prostaglandin E2 significantly increased VO2 and body temperature in all age groups. Resting plasma corticosterone concentrations were significantly elevated in the aged and adult mice, whereas injection of mIL-1 beta significantly raised plasma corticosterone concentrations in all animals. These findings indicate that the impaired febrile response of aged female C57Bl/lcrf-a(t) mice may be caused by increased concentrations and/or sensitivity to endogenous glucocorticoids. The impaired febrile responses of aged mice appear to be mediated by endogenous lipocortin-1.
Stimulation of the renin-angiotensin system by growth hormone in Lewis dwarf rats
Authors: Wyse B. , Waters M. , Sernia C.
Published: December 2017

A genetically growth hormone (GH)-deficient strain of Lewis rats was used to test the hypothesis that the actions of GH on electrolyte and fluid homeostasis are mediated by the renin-angiotensin-aldosterone system (RAAS). Dwarf rats injected with recombinant bGH (2 mg.kg-1 x day-1) for 7 days (group GH1+) and 28 days (group GH4+), respectively, were compared with saline-injected dwarf (group GH-) and normal (group N) Lewis rats. GH decreased Na+ excretion and increased renal glomerular filtration rate in dwarf rats. The dietary intake and plasma concentrations of Na+ and K+ remained unchanged. GH increased plasma insulin-like growth factor I (IGF-I) concentrations in dwarf rats (GH - = 109 +/- 9, GH1+ = 184 +/- 5, GH4+ = 189 +/- 28, N = 477 +/- 29 ng/ml plasma). Plasma angiotensinogen increased towards the levels found in normal Lewis rats (GH- = 859 +/- 38, GH1+ = 906 +/- 18, GH4+ = 1,027 +/- 19, N = 1497 +/- 80 ng angiotensin I/ml plasma); plasma renin activity increased above that of the normal Lewis (GH- = 10.2 +/- 0.6, GH1+ = 11.7 +/- 0.7, GH4+ = 16.7 +/- 2.4, N = 10.6 +/- 0.8 ng angiotensin I.ml plasma-1 x h-1). Plasma aldosterone, corticosterone, and triodothyronine concentrations were unchanged by GH treatment. Angiotensin II receptor densities in GH- rats (liver = 356 +/- 23, kidney = 228 +/- 28, adrenal = 478 +/- 58 fmol/mg protein) were upregulated by GH (GH4+ rats; liver = 573 +/- 27, kidney = 360 +/- 86, adrenal = 721 +/- 78 fmol/mg protein).(ABSTRACT TRUNCATED AT 250 WORDS)
Correlation of diacylglycerol level and protein kinase C activity in rat retina to retinal circulation
Authors: Shiba T. , Inoguchi T. , Sportsman J. R. , Heath W. F. , Bursell S. , King G. L.
Published: December 2017

The increases in diacylglycerol (DAG) level and protein kinase C (PKC) activity have been characterized biochemically and functionally in the retina and the brain of diabetic rats as well as in cultured vascular cells. PKC specific activities were increased in the membraneous fraction of retina from streptozotocin (STZ)-induced diabetic rats and the genetically determined diabetic BB rats, respectively, after 1 or 2 wk of diabetes, compared with control. The ratio of total PKC activities from membraneous and cytosol fractions was also increased in the retina of diabetic rats. With diabetes, all the isoenzymes and the total DAG level were increased in the rat retina, whereas no changes were found in the rat brain. Insulin treatment normalized plasma glucose levels and partially prevented the increases in the membraneous PKC activity and all the isoenzymes in the retina. In the retinal endothelial cells, the total DAG level and PKC specific activities are increased by 36 and 22%, respectively, in the membraneous pool when the glucose levels are changed from 5.5 to 22 mM. Activation of PKC activity and isoform beta II by the vitreal injection of phorbol dibutyrate mimicked the abnormal retinal blood circulation observed in diabetic rats (2.22 +/- 0.24 vs. 1.83 +/- 0.40 s). Thus diabetes and elevated glucose levels will increase DAG level and PKC activities and its isoenzyme specifically in vascular cells and may affect retinal hemodynamics.
Glucocorticoid receptors in PVN: interactions with NE, NPY, and Gal in relation to feeding
Authors: Tempel D. L. , Leibowitz S. F.
Published: December 2017

Norepinephrine (NE) and neuropeptide Y (NPY) potentiate carbohydrate ingestion after injection into the paraventricular nucleus (PVN), whereas injection of galanin (Gal) potentiates fat intake. The present study examines the relation between these neurochemically induced feeding behaviors and the adrenal steroids acting locally within the PVN. Results demonstrate that PVN NE- and NPY-induced carbohydrate intake is abolished by adrenalectomy surgery (ADX) and by local PVN implants of the type II receptor antagonist RU-486. Carbohydrate intake in response to PVN NE or NPY injection is unaffected by the type I antagonist RU-28318. In contrast, the stimulatory effect of PVN Gal injection on fat intake is unchanged by surgical ADX or by PVN administration of RU-486 or RU-28318, suggesting that the stimulatory action of Gal on fat ingestion occurs independently of corticosterone (Cort) and of PVN type I or type II steroid receptors. It is concluded that endogenous Cort has a permissive effect on the carbohydrate feeding responses elicited by NE and NPY in the PVN and that this interaction is mediated by type II glucocorticoid receptors within this nucleus.
Insulin resistance in the GK rat: decreased receptor number but normal kinase activity in liver
Authors: Bisbis S. , Bailbe D. , Tormo M. A. , Picarel-Blanchot F. , Derouet M. , Simon J. , Portha B.
Published: December 2017

We have previously shown that the glucose intolerance and the hyperglycemic state in the GK rat, a new spontaneous model of non-insulin-dependent (type II) diabetes without obesity, are partly accounted for by an alteration of the pancreatic B cell response. On the other hand, the hyperglycemic-hyperinsulinemic pattern in these rats suggests a decrease of response to insulin in the basal state. In the present study, in vivo insulin action was assessed in 8-wk-old GK females at basal and submaximal (euglycemic clamp) insulin levels. Overall glucose utilization (OGU), individual tissue glucose utilization (ITGU, in vivo uptake of the glucose analogue 2-deoxy-D-glucose as the relative index of glucose metabolism), as well as hepatic glucose production (GP) and liver insulin receptor properties were determined under these two conditions. The basal OGU was significantly higher in the GK females, compared with that in control Wistar females. The hyperinsulinemic-euglycemic clamp experiments indicated that peripheral insulin resistance was installed at 8 wk of age in the GK females because 1) OGU was significantly lower and 2) in some peripheral tissues (epitrochlearis muscle, periovarian, and inguinal white adipose tissues), but not all, ITGU was significantly lower compared with corresponding ITGU in control rats. In the basal state GP was significantly higher in the GK rats. At submaximal hyperinsulinemia (and euglycemia), it was less effectively suppressed than in the controls, thus demonstrating liver insulin resistance. Under both basal state and clamp condition, binding of 125I-A14-insulin to liver membranes of GK rats was significantly decreased by 20-30%.(ABSTRACT TRUNCATED AT 250 WORDS)
Growth hormone treatment of obese women for 5 wk: effect on body composition and adipose tissue LPL activity
Authors: Richelsen B. , Pedersen S. B. , Borglum J. D. , Moller-Pedersen T. , Jorgensen J. , Jorgensen J. O.
Published: December 2017

Because it has been found that growth hormone (GH) treatment of GH-deficient adults is able to reduce the total fat mass, the present study was undertaken to investigate the effect of GH treatment in obese subjects. The investigation was a double-blind placebo-controlled crossover study in which nine obese females were treated with GH (0.03 mg.kg ideal body wt-1.day-1) and placebo for 5 wk. Body composition was determined by dual-energy X-ray absorptiometry, and the quantity of intra-abdominal adipose tissue was determined by CT scan. Lipoprotein lipase (LPL) activity was determined in fat biopsies taken from the subcutaneous abdominal and gluteal region. GH treatment significantly reduced the total fat mass from 40.5 to 38.4 kg (i.e., 5% reduction of the total fat mass; P < 0.01), whereas the fat-free mass increased from 50.5 to 53.5 kg (P < 0.01). In addition, GH treatment significantly reduced the intra-abdominal adipose tissue determined by CT scan (reduction by 7 +/- 0.3%; P < 0.02). CT scan performed at the level of the femur showed a 7% reduction in adipose tissue and a 5% increase in muscle volume in the GH group (P < 0.05). Thus no clear regional differences in the GH-mediated reduction of the adipose tissue mass were observed. GH reduced the LPL activity by approximately 50% (P < 0.01) in the adipose tissue. Finally, GH treatment significantly increased the level of plasma free fatty acids (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
Activation of G protein-coupled K+ channels by dopamine in human GH-producing cells
Authors: Takano K. , Asano S. , Yamashita N.
Published: December 2017

Dopamine (DA) inhibited the secretion of growth hormone (GH) from cultured human GH-secreting adenoma cells. The mechanism of this DA effect on these cultured cells was investigated with electrophysiological techniques. Under current clamp, DA (10(-6) M) hyperpolarized the membrane and arrested Ca(2+)-dependent action potentials. Voltage clamp experiments revealed that this membrane hyperpolarization was the result of a K+ conductance increase caused by DA. The current-voltage relationship of the DA-induced K+ current showed an inward-going rectification. Application of sulpiride (10(-6) M) abolished the DA-induced K+ current, indicating that the hyperpolarization was caused by the activation of D2-like receptors. Pertussis toxin (PTX) treatment eliminated the DA-induced K+ current as well as the DA-induced inhibition of GH secretion. An intracellular application of guanosine-5'-O-(3-thiotriphosphate) (100 microM) evoked a spontaneous increase in the K+ current in the absence of an agonist, and the application of DA did not further increase conductance. Intracellular application of guanosine-5'-O-(2-thiodiphosphate) (2 mM) inhibited the DA-induced K+ current. These results indicate that the DA-induced K+ channel is coupled to a G protein. When adenosine 3',5'-cyclic monophosphate (cAMP, 100 microM) was added to the patch-pipette solution, the DA-induced K+ current was still observed, indicating that the DA-induced K+ current was not caused by an inhibition of cAMP production.(ABSTRACT TRUNCATED AT 250 WORDS)
Muscle bioenergetics in obese Zucker rats
Authors: Klein M. , Kaminsky P. , Walker P. M. , Straczek J. , Barbe F. , Duc M. , Burlet C.
Published: December 2017

The purpose of this study was to investigate the energetic metabolism in obese Zucker rats, using phosphorus nuclear magnetic resonance spectroscopy at rest and during a 2-Hz muscle stimulation and subsequent recovery. Animals were anesthetized with ketamine (150 mg/kg ip). Fed obese rats and 2-day-fasted obese rats were compared with their normally fed and 2-day-fasted lean litter mates. No differences were found between the two groups for ATP, total creatine, phosphocreatine (PCr), and intracellular pH. Starvation in lean rats resulted in a significant fall in inorganic phosphate (Pi), increased resting ADP level, and decreased PCr and ADP recovery after stimulation. The obese rats exhibited a decreased PCr/Pi and increased ADP at rest and a decreased PCr resynthesis and ADP metabolization rate after stimulation. Muscle stimulation in fasted obese rats induced higher PCr depletion and more pronounced acidosis. These results suggest an in vivo mitochondrial metabolism dysfunction in fasted lean as well as in fed and fasted obese rats.
Activation of PI 3-kinase in 3T3-L1 adipocytes by association with insulin receptor substrate-1
Authors: Lamphere L. , Carpenter C. L. , Sheng Z. F. , Kallen R. G. , Lienhard G. E.
Published: December 2017

Insulin treatment of adipocytes causes the rapid phosphorylation of the insulin receptor substrate-1 (IRS-1) on tyrosine. The phosphotyrosine [Tyr(P)] form of IRS-1 then complexes with the enzyme phosphatidylinositol (PI) 3-kinase. In this study, we have investigated the effect of this association on PI 3-kinase activity in 3T3-L1 adipocytes. Insulin stimulated cytosolic PI 3-kinase activity about sevenfold. This stimulation was maximal after 1 min of exposure of cells to insulin, persisted for at least 1 h, and occurred over the range of insulin concentrations that saturate its receptor. By means of immunoprecipitation of IRS-1, it was shown that virtually all of the enhanced activity was due to PI 3-kinase complexed with IRS-1. Moreover, the purified Tyr(P) form of IRS-1, either isolated from 3T3-L1 adipocytes or obtained by phosphorylation of the recombinant protein with the insulin receptor, markedly stimulated the activity of purified rat liver PI 3-kinase. These results show that the association of Tyr(P) IRS-1 with PI 3-kinase activates the enzyme and thereby can explain the elevation of PI 3,4-bisphosphate and PI 3,4,5-trisphosphate in vivo observed upon treatment of adipocytes with insulin.
Experimental reliability of the doubly labeled water technique
Authors: Goran M. I. , Poehlman E. T. , Danforth E.
Published: December 2017

The experimental reliability of measuring CO2 production rates (rCO2) with the doubly labeled water (DLW) technique was assessed in five young healthy men (23 (DLW) technique was assessed in five young healthy men (23 +/- 4 yr; 66.1 +/- 4.6 kg). To minimize the confounding effects of fluctuations in physical activity and eating patterns on variation in energy expenditure, the subjects lived under sedentary living conditions by confinement to their own room at a Clinical Research Center and were maintained on a fixed and known level of energy intake. rCO2 was determined in duplicate over two identical 9-day study periods after separate loading doses of deuterium and oxygen-18. Turnover rates were determined from multipoint sampling to reduce error from analytical uncertainty. Dilution spaces were determined by both the intercept and plateau methods. The average experimental variation for rCO2 estimates was approximately +/- 8.5% and was not significantly different among three published calculation models that differ in their assumptions regarding the relationship between the dilution spaces of deuterium and oxygen-18. The experimental reliability of +/- 8.5% exceeds theoretical values generated from calculations based on propagation of error from analytical uncertainty. Between subjects, the experimental variation ranged from 1 to 21%, and the half-width of the 95% confidence interval for the precision of rCO2 estimates was high (+/- 12 mol/day) relative to the mean reported value of approximately 16 mol/day.(ABSTRACT TRUNCATED AT 250 WORDS)
Coordinated regulation of hepatic glycogen formation in perfused rat liver by glucose and lactate
Authors: Zhang Z. , Radziuk J.
Published: December 2017

Lactate has been found to enhance the formation of glycogen from both glucose and lactate as substrate (Z. Zhang and J. Radziuk. Biochem. J. 280: 415–419, 1991). To evaluate the relative importance of its role as substrate and regulatory factor, a dual dose-response evaluation was done by adding variable amounts of glucose and lactate to the medium in a recirculating perfused rat liver preparation. Nine groups of perfusions were performed utilizing three different levels of carbon infusion into the system: 0.25, 1.0, and 2.0 mg/min. These levels of carbon infusion were further subdivided into different relative amounts of glucose and lactate. Lactate uptake by the perfused liver was linearly related with net glucose output, regardless of the glucose concentrations. In contrast to this, the effect of lactate uptake on the rate of glycogen synthesis is saturable. Moreover, the rate of glycogen formation at which this saturation occurs is dependent only on the mean perfusate glucose concentration. The highest amount of glycogen formed in a 2-h period was 50 +/- 7 mg and the lowest 3.4 +/- 0.3 mg. A family of dose-response curves was generated describing this dual dependence of glycogen formation (both direct and gluconeogenetic pathways) on lactate and glucose.
Effects of chronic hypercortisolemia on carbohydrate metabolism during insulin deficiency
Authors: Goldstein R. E. , Cherrington A. D. , Reed G. W. , Lacy D. B. , Wasserman D. H. , Abumrad N. N.
Published: December 2017

This study was undertaken to further investigate the effect of acute selective insulin deficiency on glycogenolysis and gluconeogenesis occurring during chronic physiological hypercortisolemia in conscious overnight fasted dogs. After an 80-min tracer and dye equilibration period and a 40-min basal period, selective insulin deficiency was created during the 180-min experimental period by infusing somatostatin peripherally (0.8 micrograms.kg-1.min-1) with basal replacement of glucagon intraportally (0.65 ng.kg-1.min-1). In the cortisol group (n = 5), a continuous infusion of hydrocortisone (3.5 micrograms.kg-1.min-1) was begun 5 days before the experiment. In the saline group (n = 5), there was no infusion of cortisol. [3-3H]glucose, [U-14C]alanine, and indocyanine green dye were used to assess glucose production and gluconeogenesis using tracer and arteriovenous difference techniques. During selective insulin deficiency in the saline group, the arterial plasma glucose level (Glc) increased from 109 +/- 2 to 285 +/- 19 mg/dl; glucose production increased from 2.7 +/- 0.2 to 4.5 +/- 0.3 mg.kg-1.min-1. Gluconeogenic efficiency and conversion of alanine to glucose (Conv) increased by 300 +/- 55 and 356 +/- 67%. During selective insulin deficiency in the cortisol group, Glc increased from 117 +/- 3 to 373 +/- 50 mg/dl; glucose production increased from 3.3 +/- 0.5 to 6.9 +/- 0.7 mg.kg-1.min-1. Gluconeogenic efficiency and Conv increased by 268 +/- 41 and 393 +/- 75%, respectively. The maximal glycogenolytic rate increased significantly more in the cortisol group than in the saline group, accounting for the difference in glucose production. These results suggest that, even during chronic hypercortisolemia, acute insulin deficiency has more pronounced effects on glycogenolysis than gluconeogenesis.
In vivo measurement of fatty acids and cholesterol synthesis using D2O and mass isotopomer analysis
Authors: Lee W. N. , Bassilian S. , Ajie H. O. , Schoeller D. A. , Edmond J. , Bergner E. A. , Byerley L. O.
Published: December 2017

The synthesis of palmitate, stearate, and cholesterol in liver and nervous tissues (brain, cord, and nerve) of Sprague-Dawley rats was determined using deuterated water (D2O) and mass isotopomer analysis. Rats were given 4% deuterium in their drinking water after each receiving an intraperitoneal priming dose. Animals were killed at 1, 2, 4, and 8 wk for deuterium enrichment in body water and determination of mass isotopomer distribution in lipids from various tissues. In 1 wk, the enrichment in the body water reached a plateau of 2.6%, which is 65% of that in the drinking water. We observed the maximum incorporation number (N) in all lipids to be higher than those previously observed, being 22, 24, and 30 for liver palmitate, stearate, and cholesterol, respectively, and N may vary among tissues. Using a single exponential model, we found the half-time (t1/2) and the plateau levels of the newly synthesized lipids of the nervous tissues (t1/2 values ranging from 5 to 28 days) to be different from those of the liver (t1/2 values < or = 4 days) in this relatively long-term study. Mass isotopomer distribution analysis and D2O can be used not only to quantitate the replacement rate of many lipids in various compartments but may also be used to elucidate the tissue-specific synthetic pathways from N.
Canine liver releases neuropeptide Y during sympathetic nerve stimulation
Authors: Taborsky G. J. , Beltramini L. M. , Brown M. , Veith R. C. , Kowalyk S.
Published: December 2017

To determine whether the liver or gut releases neuropeptide Y (NPY) from their sympathetic nerves, we performed bilateral thoracic sympathetic nerve stimulation (BTSNS) in halothane-anesthetized dogs and calculated gut and liver NPY spillover. BTSNS markedly increased hepatic NPY spillover (delta = +32 +/- 8 ng/min) and arterial NPY concentration (delta = +220 +/- 56 pg/ml), despite no effect on gut NPY spillover (delta = +8 +/- 7 ng/min). To determine the liver's contribution to this increase of circulating NPY, hepatic nerves were selectively stimulated (HNS). Liver NPY spillover increased markedly (delta = +114 +/- 42 ng/min, P < 0.025) during HNS, causing a large increase of arterial NPY (delta = +586 +/- 237 pg/ml, P < 0.025). Using this ratio of liver spillover to arterial increments of NPY, we calculated that the liver makes a major contribution (70%) to circulating NPY levels during BTSNS. The predominant form of canine NPY coeluted with synthetic [Met17]NPY and the minor form of canine NPY coeluted with the oxidized form of [Met17]NPY on high-performance liquid chromatography. We therefore conclude that dog NPY is likely [Met17]NPY and that the liver, rather than the gut, is a major source of circulating NPY during sympathetic nerve stimulation and perhaps stress.
Kinetic model of the response of precursor and mature rat hepatic mRNA-S14 to thyroid hormone
Authors: Goumaz M. O. , Schwartz H. , Oppenheimer J. H. , Mariash C. N.
Published: December 2017

We found in preliminary experiments that multiple daily injections of triiodothyronine (T3) resulted in an apparent prolongation in the half time (t1/2) of mRNA-S14 decay. To appropriately interpret these observations, we developed a mathematical model of the fluctuations of mRNA-S14 and its nuclear precursor after a single injection or multiple daily injections of T3. The model parameters include 1) the effect of plasma protein binding and metabolic clearance rates on receptor-bound nuclear T3, 2) the threefold circadian variation in mRNA-S14, 3) a 12-min t1/2 for the nuclear precursor and a 1.5-h t1/2 for the mature mRNA-S14, 4) previously derived relationships between the level of plasma T3 and nuclear occupancy, and 5) direct proportionality between nuclear transcription of the S14 gene and T3 nuclear occupancy. The model faithfully predicted the excursions of the mature mRNA-S14 and its nuclear precursor. Nuclear retention of T3 and the effects of circadian variation on S14 gene transcription explain the apparent prolongation in the t1/2 of decay of mature mRNA. Our findings illustrate the feasibility of incorporating parameters at the molecular level into a comprehensive kinetic analysis of hormone action.
Effect of adrenalectomy on glucose tolerance and lipid metabolism in gold-thioglucose obese mice
Authors: Blair S. C. , Caterson I. D. , Cooney G. J.
Published: December 2017

The effect of adrenalectomy (ADX) on body weight, lipogenesis, and glucose tolerance was investigated in mice made obese by a single intraperitoneal injection of gold-thioglucose (GTG). Five weeks after ADX the weight of GTG-obese mice was significantly decreased (GTG-obese+sham-ADX: 39.8 +/- 0.8 g; GTG-obese+ADX: 27.6 +/- 1.1 g; P < 0.05). ADX also reduced serum glucose (GTG-obese+sham-ADX: 16.5 +/- 0.6 mmol/l; GTG-obese+ADX: 10.8 +/- 0.5 mmol/l; P < 0.05) and serum insulin concentrations (GTG-obese+sham-ADX: 197 +/- 36 microU/ml; GTG-obese+ADX: 38 +/- 7 microU/ml; P < 0.05) of fed GTG-obese mice and greatly improved glucose tolerance. ADX lowered liver glycogen content and reduced the fatty acid content of liver, epididymal white adipose tissue (WAT), and interscapular brown adipose tissue (BAT) of fed GTG-obese mice. Lipid synthesis in liver and WAT of GTG-obese mice was decreased by ADX, but lipogenesis in BAT was increased, possibly to provide substrate for increased thermogenesis in this tissue. Effects of ADX on metabolism were not confined to GTG-injected mice, as ADX also reduced body weight and altered the glucose tolerance of age-matched control mice. ADX increased lipid synthesis in liver, WAT, and BAT of fed control mice without an increase in lipid deposition, indicating that there was increased lipid turnover in these lipogenic tissues of ADX mice. ADX reduced the fasting blood glucose concentration of both control and GTG-obese mice to a level below that of sham-ADX control mice (sham-ADX control: 6.0 +/- 0.4 mM; ADX control: 2.9 +/- 0.5 mM; ADX GTG-obese: 3.3 +/- 0.2 mM).(ABSTRACT TRUNCATED AT 250 WORDS)
Stimulation of trabecular bone formation by insulin-like growth factor I in adult ovariectomized rats
Authors: Mueller K. , Cortesi R. , Modrowski D. , Marie P. J.
Published: December 2017

Although in vitro experiments indicate that insulin-like growth factor I (IGF-I) is an anabolic hormone in bone cell metabolism, the effects of IGF-I in vivo on bone formation are unclear. We thus investigated whether IGF-I is able to stimulate bone formation in adult rats with established osteopenia induced by ovariectomy (OVX). IGF-I was administered at daily doses of 0.05, 0.2, and 0.8 mg/kg for 3 wk. OVX induced a marked osteopenia in femur and tibia. Administration of IGF-I increased trabecular bone mass with a maximal effect at 0.2 mg/kg. The same dose stimulated bone formation, as revealed by an increase in osteoid surface, osteoblast surface, triple tetracycline-labeled surface, and bone formation rate. The mineral apposition rate was equally stimulated at all doses. At the highest dose, IGF-I increased osteoclast surface and osteoclast number. These data indicate that, in the adult OVX rat, IGF-I stimulates bone formation and increases trabecular bone volume at medium doses and enhances the histological indexes of bone resorption at high doses.
Effects of exercise on protein synthesis and myosin heavy chain gene expression in hypothyroid rats
Authors: Katzeff H. L. , Ojamaa K. M. , Klein I.
Published: December 2017

Hypothyroidism suppresses muscle growth and alters myosin heavy chain (MHC) gene expression. To study the role of thyroid hormones in exercise-induced muscle growth and protein synthesis, we measured skeletal and cardiac muscle protein synthesis and MHC gene expression in hypothyroid rats allowed to exercise voluntarily. Female Sprague-Dawley rats (200-210 g) were separated into four groups for 28 days of treatment: control, hypothyroid (TX), hypothyroid plus running-wheel exercise (TX+Ex), and hypothyroid plus 25% overfed (TX+OF). Fractional protein synthesis rates (% incorporation/day) were measured using [3H]phenylalanine incorporation 10 min postinjection. The heart weight-to-body weight ratios of the TX and the TX+OF groups showed marked cardiac atrophy over the 28-day period (2.76 +/- 0.12 and 2.50 +/- 0.22 vs. 3.37 +/- 0.18 mg/g, respectively; P < 0.01). However, the TX+Ex group prevented heart, gastrocnemius, and soleus muscle atrophy over the same time period. Heart, gastrocnemius, and soleus muscles had markedly suppressed protein synthesis rates in the TX and TX+OF groups vs. the euthyroid controls (mean fall -72%; P < 0.01, analysis of variance). However, exercise increased protein synthesis rate by 50% (P < 0.05) compared with TX alone in all three muscle groups. Exercise did not modify hypothyroid-induced alterations of cardiac myosin isoform expression. Exercise-mediated effects on skeletal and cardiac muscle growth but not cardiac MHC gene expression appear to be independent of thyroid hormones.
Human placenta makes extracellular glutathione peroxidase and secretes it into maternal circulation
Authors: Avissar N. , Eisenmann C. , Breen J. G. , Horowitz S. , Miller R. K. , Cohen H. J.
Published: December 2017

Extracellular glutathione peroxidase (eGPX) is a selenoglycoprotein distinct from cellular glutathione peroxidase (cGPX). The cDNA for eGPX has recently been cloned from human placenta. To determine whether human placenta makes both cGPX and eGPX and secretes eGPX, we used specific immunoprecipitations of 75Se metabolically labeled proteins from full-term placental explants in culture and perfused placental lobules. Placental explants and metabolically active, dually perfused placental lobules synthesized and contained both cGPX and eGPX and secreted eGPX. Perfused tissue secreted eGPX into the maternal but not into the fetal perfusate. In situ hybridizations using antisense and sense eGPX riboprobes were performed on sections of first-, second-, and third-trimester placentas. In the first-trimester placenta, transcripts were localized predominantly to cytotrophoblast cells, whereas in the full-term placenta syncytiotrophoblast cells and stromal cells but not fetal endothelial cells expressed eGPX mRNA. It is concluded that human placenta synthesizes both cGPX and eGPX and secretes eGPX into the maternal circulation, consistent with the location of the eGPX mRNA.
Precursor pools of protein synthesis: a stable isotope study in a swine model
Authors: Baumann P. Q. , Stirewalt W. S. , O'Rourke B. D. , Howard D. , Nair K. S.
Published: December 2017

The accuracy of using other free pools in lieu of tRNA for calculation of tissue protein synthesis in liver (L), skeletal muscle (SM), and heart (H) was assessed in six adult miniature swine using L-[1-13C]leucine and L-[ring-2H5]phenylalanine as tracers. L leucyl-tRNA enrichment was higher than arterial plasma leucine and ketoisocaproate (KIC) enrichments, and L phenylalanyl-tRNA enrichment was higher than arterial phenylalanine enrichment (P < 0.05). No such differences were noted in SM and H. Leucyl- and phenylalanyl-tRNA enrichments in L were best predicted by the respective amino acid enrichments in tissue fluid [TF; Leu: slope (m) = 0.954 +/- 0.035; Phe: m = 1.011 +/- 0.032] using linear regression analysis to determine the accuracy of the prediction, whereas plasma phenylalanine reasonably predicted phenylalanyl-tRNA (artery: m = 0.821 +/- 0.032; vein: m = 0.947 +/- 0.135). In SM, plasma KIC (artery: m = 0.846 +/- 0.046; vein: m = 0.881 +/- 0.043) and TF leucine (m = 0.788 +/- 0.034) predicted leucyl-tRNA with high accuracy. In H tissue, TF (m = 0.991 +/- 0.044) was the best predictor of leucyl-tRNA enrichment, whereas arterial phenylalanine (m = 0.912 +/- 0.015) was the most reliable predictor of phenylalanyl-tRNA enrichment. The relationships between aminoacyl-tRNA and other free pools in the same species under the same study conditions differ in different tissues. Use of KIC in lieu of leucyl-tRNA for calculating muscle protein synthesis is supported by this study.
Expression of the genes for insulin-like growth factors and their receptors in bone during skeletal growth
Authors: Bikle D. D. , Harris J. , Halloran B. P. , Roberts C. T. , Leroith D. , Morey-Holton E.
Published: December 2017

Insulin-like growth factors (IGF) are important regulators of skeletal growth. To determine whether the capacity to produce and respond to these growth factors changes during skeletal development, we measured the protein and mRNA levels for IGF-I, IGF-II, and their receptors (IGF-IR and IGF-IIR, respectively) in the tibia and femur of rats before and up to 28 mo after birth. The mRNA levels remained high during fetal development but fell after birth, reaching a nadir by 3-6 wk. This fall was most pronounced for IGF-II and IGF-IIR mRNA and least pronounced for IGF-I mRNA. However, after 6 wk, both IGF-I and IGF-IR mRNA levels recovered toward the levels observed at birth. In the prenatal bones, the signals for the mRNAs of IGF-II and IGF-IIR were stronger than the signals for the mRNAs of IGF-I and IGF-IR, although the content of IGF-I was three- to fivefold greater than that of IGF-II. IGF-II levels fell postnatally, whereas the IGF-I content rose after birth such that the ratio IGF-I/IGF-II continued to increase with age. We conclude that, during development, rat bone changes its capacity to produce and respond to IGFs with a progressive trend toward the dominance of IGF-I.
IGF-I and IGF-binding protein gene expressions in spontaneous dwarf rat
Authors: Nogami H. , Watanabe T. , Kobayashi S.
Published: December 2017

Effects of growth hormone (GH) and fasting on hepatic expressions of insulin-like growth factor I (IGF-I) and IGF-I-binding protein (IGFBP)-1, -2, -3, and -4 were examined in spontaneous dwarf rats (SDR), which completely and specifically lack GH among pituitary hormones. The hepatic expressions of mRNA encoding IGF-I and IGFBP-3 were reduced and IGFBP-1 mRNA was elevated in the SDR. Both chronic and acute administration of GH restored these changes, indicating the association of GH but not other pituitary hormones with hepatic expressions of these genes. In addition, the present examination revealed that mRNA level of IGFBP-2 was elevated in SDR, which could not be attenuated by exogenous GH, and that GH may not be directly relevant to the regulation of hepatic IGFBP-4 expression. Fasting for 2 days reduced IGF-I mRNA level and increased IGFBP-2 mRNA level in the SDR, as well as in the normal rat, suggesting the presence of factors other than reduced serum GH responsible for fasting-induced alteration in the expression of these mRNAs. On the other hand, fasting resulted in little change or even a reduction of IGFBP-1 mRNA level in the SDR.
Evidence for a sustained genetic effect on fat storage capacity in cultured adipose cells from Zucker rats
Authors: Briquet-Laugier V. , Dugail I. , Ardouin B. , Le Liepvre X. , Lavau M. , Quignard-Boulange A.
Published: December 2017

Using mature adipocytes and preadipocytes from genetically obese Zucker rats, we investigated the cells' ability to maintain abnormal fat storage capacity when withdrawn from their in vivo environment. Long-term adipocyte cultures from obese rats displayed an increase in both glucose consumption (GC) and enzyme activities, including fatty acid synthase (4-fold), glycerol-3-phosphate dehydrogenase (4.5-fold), lipoprotein lipase (LPL; 6-fold), and malic enzyme (2.5-fold). Fully differentiated obese predipocytes exhibited a twofold increase in these enzyme activities, together with higher glucose metabolism. In obese cells, LPL mRNA was increased in both adipocytes (6-fold) and differentiated preadipocytes (2-fold). Insulin mediated an increase in GC and lipogenic enzymes in both adipocytes and preadipocytes regardless of the genotype; this effect was more marked in obese cells. Examining cultured adipocytes from rats fed a high-fat diet, we showed that the nutritional effect upon GC and lipogenic enzymes was abolished after culture. These results demonstrated that fatty mutation may be intrinsically expressed in prolonged cultured mature adipocytes and in newly differentiated adipocytes.
Effect of growth hormone on hepatic glucose and insulin metabolism after oral glucose in conscious dogs
Authors: Okuda Y. , Pena J. , Chou J. , Field J. B.
Published: December 2017

This study examined the effect of growth hormone (GH) on hepatic glucose metabolism and on the fractional extraction of insulin and glucagon after oral glucose administration. GH treatment [canine GH (0.75 mg/day for 7 days)] significantly increased basal portal vein and hepatic artery flow (P < 0.01 compared with pre-GH treatment). After GH treatment and after oral glucose, glucose levels significantly exceeded those before GH at 100 and 120 min in arterial and portal vein plasma and 120 min in the hepatic vein. The net hepatic uptake of glucose was similar before and after GH treatment. The increment of net nonhepatic splanchnic insulin balance above basal was 131 +/- 31 mU.kg-1.3 h-1 before and 272 +/- 46 mU.kg-1.3 h-1 after GH treatment (P < 0.05). An increase in fractional hepatic extraction of insulin occurred before GH treatment and was significantly greater at 60 min. In summary, despite the increased insulin content after GH administration, there was no change in hepatic uptake of glucose, indicating that the liver was also the site of insulin resistance.
Enhanced adrenal renin and aldosterone biosynthesis during sodium restriction in TGR (mREN2)27
Authors: Rubattu S. , Enea I. , Ganten D. , Salvatore D. , Condorelli G. , Condorelli G. , Russo R. , Romano M. , Gigante B. , Trimarco B. , et al.
Published: December 2017

The aim of the study was to investigate the relationships between tissue renin and the steroid production in the adrenal cortex during dietary sodium restriction in the transgenic rat (TGR) (mREN2)27. Thus the effects of a 1-wk low-sodium intake (0.04% NaCl) were studied in 5-wk-old male TGR (n = 33, systolic blood pressure = 151 +/- 3 mmHg) and in 24 age- and sex-matched outbred normotensive Sprague-Dawley (SD) rats. Measurements of plasma and tissue hormones were obtained at 0, 4, and 7 days of a low-sodium diet. Sodium restriction caused sustained increases of adrenal renin activity (from 28.5 +/- 3.5 to 87.5 +/- 4.5 ng.mg protein-1.h-1 on day 7) and of adrenal renin mRNA (+63 +/- 13 and +43 +/- 7% on days 4 and 7, respectively), whereas plasma renin activity (from 3.3 +/- 0.3 to 4.4 +/- 0.6 ng.ml-1.h-1) and renal renin activity (from 0.85 +/- 0.25 to 0.7 +/- 0.4 microgram.mg protein-1.h-1) did not change. The stimulation of the adrenal renin-angiotensin system was associated with a large increase of the aldosterone synthase cytochrome P-450 mRNA (+165 +/- 35 and +184 +/- 44%, on days 4 and 7) and of plasma aldosterone levels (from 125 +/- 32 to 338 +/- 59 pg/ml, P < 0.01). In SD rats, in spite of a more consistent increase in renal and circulating renin, mineralocorticoid production did not increase significantly. These results demonstrate that the exaggerated biosynthesis of aldosterone in TGR during sodium restriction is associated with an activation of renin in the adrenal cortex but not in the kidney.
Effect of central amiloride infusion on mineralocorticoid hypertension
Authors: Gomez-Sanchez E. P. , Gomez-Sanchez C. E.
Published: December 2017

There is strong evidence from different types of studies, including the discrete infusion of agonists and antagonists and ablation of specific brain areas or transmitter-type neurons, that mineralocorticoids, in excess, act in the brain to elevate blood pressure. Aldosterone enhances the entry of Na+ through amiloride-sensitive Na+ channels in some mineralocorticoid-sensitive transport epithelial cells. To define possible cellular mechanisms involved in central mineralocorticoid action, benzamil, an amiloride analogue with selective affinity for the Na+ channel, was continuously infused intracerebroventricularly in three mineralocorticoid-dependent hypertension models in Sprague-Dawley rats, the continuous subcutaneous infusion of aldosterone, the intracerebroventricular infusion of aldosterone, and the ingestion of carbenoxolone, a synthetic licorice analogue. The intracerebroventricular infusion of 0.3 and 0.5 micrograms/h of benzamil, doses that did not have an adverse effect on growth and that had no effect on the blood pressure when infused subcutaneously, prevented the increase in blood pressure in these models. The infusion of these levels of benzamil had no effect on urine volume even in those animals in which it prevented an increase in blood pressure. These data suggest that the central effects of mineralocorticoids on blood pressure are mediated, at least in part, by the effects of mineralocorticoids on amiloride-sensitive sodium transport.
Convolution: a method for data analysis in perifusion systems
Authors: Brand R. M. , Midgley A. R. , Williams W. J.
Published: December 2017

Considerable interest has developed in defining how imposed stimuli effect dynamic changes in cellular metabolism. We have developed a miniature perifusion system that can reveal alterations in extracellular protons within seconds after application of metabolic perturbants. This perifusion system contains two pH sensors: one before the cells records changes between medium and test solutions, and one, located just past the cells, records these alterations plus cellular modifications. Because distortion occurs as chemicals pass through perifusion systems, the shape of pH changes induced by switching from medium to test solutions is different at each electrode. This study describes and validates convolution to correct this distortion. Data from HeLa cells exposed to the metabolic uncoupler of oxidative phosphorylation carbonyl cyanide m-chlorophenyl hydrazone have been analyzed with convolution. Cellular response to oxidative phosphorylation removal is comprised of multiple components, is consistent with a rapid uncoupling, and is followed by cellular adaptation. Therefore convolutional analysis can provide an important adjunct to the analysis of data acquired by perifusion and can provide new insights into cellular responsiveness and metabolism.
Thyroxine and 3,5,3'-triiodothyronine production, metabolism, and distribution in pregnant rat near term
Authors: Versloot P. M. , Gerritsen J. , Boogerd L. , Schr oder-van der Elst J. P. , Van Der Heide D.
Published: December 2017

In the pregnant rat near term thyroxine (T4) and 3,5,3'-triiodothyronine (T3) concentrations are lower in plasma and extrathyroidal tissues, except T3 in the brain. To study the changes in T4 and T3 kinetics a bolus injection of [125I]T4 and [131I]T3 was administered to nonpregnant controls and rats 14 and 19 days pregnant. Physiological parameters of the production, interpool transport, distribution, and metabolism of T4 and T3 were estimated by means of a three-compartment model. The production and partition of T4 remained unchanged during pregnancy. The total distribution volume of T4 was enlarged. On day 19 the plasma clearance rate was doubled, and transport to the fast pool was more than tripled. The rate of production of T3 was slightly diminished. The plasma clearance rate was increased, but no changes were found in the interpool transport rates. These results suggest that in the pregnant rat near term the increased transport of T4 is responsible for the distribution of the available T4 between the maternal and the fetal compartment.
Calcitriol therapy and calcium-regulated PTH secretion in patients with secondary hyperparathyroidism
Authors: Ramirez J. A. , Goodman W. G. , Belin T. R. , Gales B. , Segre G. V. , Salusky I. B.
Published: December 2017

Calcitriol lowers serum parathyroid hormone (PTH) levels in patients with secondary hyperparathyroidism, but its effect on calcium-regulated PTH release remains controversial. Thus 11 patients with secondary hyperparathyroidism underwent dynamic tests of parathyroid function before and after 4 mo of intermittent calcitriol therapy. Serum calcitriol levels rose from 8 +/- 1 to 55 +/- 9 (SE) pg/ml, P < 0.01, serum total and ionized calcium levels increased, and serum PTH levels decreased from 584 +/- 154 to 154 +/- 31 pg/ml, P < 0.05. The maximum increase in serum PTH during hypocalcemia did not differ before (248 +/- 78 pg/ml) or after (280 +/- 100 pg/ml) treatment, but the increase in PTH, expressed as a percentage of preinfusion values, was greater after treatment (329 +/- 73 vs. 132 +/- 10%, P < 0.05). The decreases in serum PTH during calcium infusions did not differ before (70 +/- 5%) or after (73 +/- 5%) therapy, and the set point for PTH release did not change (1.20 +/- 0.03 vs. 1.23 +/- 0.01 mmol/l, not significant). Calcitriol modifies PTH secretion during hypocalcemia in secondary hyperparathyroidism without affecting the set point for PTH release; although calcitriol lowers serum PTH levels, it may also restore the secretory reserve of hyperplastic parathyroid tissues during hypocalcemia.