bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of intravenous injection of IL-1 beta on PGE2 levels in several brain areas as determined by microdialysis
Authors: Komaki G. , Arimura A. , Koves K.
Published: December 2017

Using perfusion microdialysis, the effect of intravenous injection of recombinant human interleukin-1 beta (IL-1 beta) on prostaglandin E2 (PGE2) release into the interstitial fluid of the organum vasculosum laminae terminalis (OVLT), medial preoptic area (MPOA), paraventricular nucleus (PVN), dorsal hippocampus (HPC), and cerebrospinal fluid (CSF) of the lateral ventricle (LV) was examined. Artificial CSF was perfused through the dialysis probe and collected at 20-min intervals before and after IL-1 beta (1 microgram/100 g body wt iv). The results showed that 1) IL-1 beta induced a significant increase in PGE2 levels in the OVLT and the medial part of the MPOA in the first 20 min, which is more rapid and to a greater extent than that in PVN, HPC, and LV; 2) inclusion of indomethacin in the perfusate abolished the IL-1 beta-induced PGE2 response in the OVLT, but a suppressive effect in the PVN was insignificant. These results suggest that OVLT may be a gate for the entry of blood-borne IL-1 beta into the brain and a site where the initial biochemical response to IL-1 beta occurs.
ACTH and cortisol responses to sequential CRF injections in fetal sheep
Authors: Kerr D. R. , Castro M. I. , Rawashdeh N. M. , Rose J. C.
Published: December 2017

To determine whether an initial ovine corticotropin-releasing factor (oCRF) injection modifies adrenocorticotropic hormone (ACTH) and cortisol responses to a second injection and to establish whether the effect changes throughout gestation, we studied chronically cannulated fetal lambs of 103-113 and 133-137 days gestation. Experimental groups underwent an injection (500 ng/kg iv) of oCRF, arterial blood sampling for 6 h, then a similar oCRF injection followed by sampling. In control studies, vehicle was the initial injection. After the first oCRF injection, plasma cortisol levels went from 1.7 +/- 0.4 to 9.5 +/- 5.2 (SE) ng/ml ("immature") and from 22.3 +/- 4.9 to 52.5 +/- 5.8 ng/ml ("mature"), remaining elevated for 6 h. In immature fetuses, the first oCRF injection did not alter the ACTH response to a second injection. Cortisol increases were reduced. In mature animals, ACTH and cortisol response to oCRF were eliminated by prior oCRF. Thus a large increase in cortisol after oCRF in mature fetuses is associated with inhibition of the ACTH response to a second oCRF injection, whereas in immature animals a small increase in cortisol after the first oCRF injection is not.
Hypertension in insulin-resistant Zucker obese rats is independent of sympathetic neural support
Authors: Zemel M. B. , Peuler J. D. , Sowers J. R. , Simpson L.
Published: December 2017

We have previously reported that insulin-resistant Zucker obese rats exhibit hypertension associated with impaired vascular smooth muscle (VSM) Ca2+ transport and proposed that this results from failure of insulin to regulate VSM Ca2+ transport in insulin resistance. However, hypertension in insulin-resistant states is generally attributed to hyperinsulinemia, with a consequent stimulation of sympathetic neural activity. Accordingly, the present study was conducted to determine whether the hypertension observed in Zucker obese rats compared with their lean controls was dependent on either increased sympathetic neural activity or exaggerated vascular reactivity. Intra-arterial blood pressure responses to ganglionic blockade with Ecolid (chlorisondamine chloride) and to graded intravenous injections of angiotensin II and norepinephrine were compared in 6- to 8-wk-old male Zucker rats and their lean controls (n = 10/group). The obese rats exhibited significant hypertension before ganglionic blockade (P less than 0.001), and this difference was largely sustained during ganglionic blockade (P less than 0.005). Furthermore, the obese rats exhibited greater pressor sensitivity to both angiotensin II and to norepinephrine during ganglionic blockade (P less than 0.01). Thus enhanced pressor sensitivity, independent of sympathetic neural activity, appears to support hypertension in Zucker obese rats.
Inhibition of phosphatidylinositol synthase by glucose in human retinal pigment epithelial cells
Authors: Nakamura J. , Del Monte M. A. , Shewach D. , Lattimer S. A. , Greene D. A.
Published: December 2017

A series of interrelated biochemical and functional defects, induced by hyperglycemia, associated with intracellular depletion of D-myo-inositol, and corrected by aldose reductase inhibitors, have been ascribed to abnormal phosphoinositide metabolism in several tissues prone to diabetic complications. However, reductions in tissue D-myo-inositol content are not universally found in complications-prone diabetic tissues, and direct mass-action effects of cellular D-myo-inositol depletion on the critical CDPdiacylglycerol-inositol 3-phosphatidyltransferase (PI synthase; EC 2.7.8.11) step have never been shown conclusively in relevant cells. The studies reported here simultaneously estimated the chemical mass of CDP diglyceride by equilibrium labeling with 5-[3H]cytidine and phosphoinositide biosynthesis by the incorporation of [32P]orthophosphate into phosphoinositide. This was done to assess the degree of inhibition of PI synthase under various degrees of D-myo-inositol depletion and sorbitol accumulation induced by glucose and other metabolic manipulations in cultured human retinal pigment epithelial cells, a new in vitro model for diabetic complications. The results suggest that sorbitol accumulation limits the PI synthase reaction in these cells by selectively depleting specific intracellular pools of D-myo-inositol and/or by possible independent effects of sorbitol on PI synthase.
Modulation of prostaglandin E2 synthesis in rat skeletal muscle
Authors: Turinsky J. , O'Sullivan D. M. , Bayly B. P.
Published: December 2017

The effect of muscle denervation, inhibitors of protein synthesis, G proteins, and sphingolipids on prostaglandin E2 (PGE2) release by rat soleus muscle in vitro was investigated. To assess the effect of muscle denervation, the sciatic nerve in one hindlimb of rats was interrupted, and soleus muscles from the denervated hindlimb and the contralateral sham (control) hindlimb were excised 1-5 days after surgery. Compared with corresponding sham muscles, PGE2 release by denervated muscles was increased 56, 230, and 435% at 1, 3, and 5 days after denervation, respectively. Protein synthesis inhibitors cycloheximide (10 microM) and puromycin (10 microM) lowered PGE2 release by sham and denervated muscles 62-80%. The release of PGE2 by sham and denervated muscles was not altered by pertussis toxin (1 microgram/ml) but was inhibited 30-51% by AlF4-. Addition of 100 microM guanosine 5'-O-(3-thiotriphosphate) to saponin-permeabilized sham and denervated muscles had only a moderate, if any, stimulatory effect on PGE2 release. This effect was not counteracted by 1 mM guanosine 5'-O-(2-thiodiphosphate). Increasing muscle ceramide concentration by incubation with sphingomyelinase (100 mU/ml) increased PGE2 release by sham and denervated muscles 43 and 157%, respectively. Because degradation of ceramides yields sphingosine, the effect of sphingosine was also tested. Sphingosine (25 microM) increased PGE2 release by sham and denervated muscles 139 and 187%, respectively, without affecting muscle viability, as assessed by the release of lactate dehydrogenase. The data indicate that muscle denervation, treatment with sphingomyelinase, and sphingosine stimulate, whereas inhibitors of protein synthesis inhibit PGE2 synthesis by muscle.
Effects of growth hormone on growth and muscle Na(+)-K+ pump concentration in K(+)-deficient rats
Authors: Dorup I. , Flyvbjerg A. , Everts M. E. , Orskov H.
Published: December 2017

K(+)-deficient rats and control rats were injected for 16 days with saline or human growth hormone (hGH, 200 micrograms/day). hGH treatment of K(+)-deficient rats resulted in increased weight gain and soleus muscle weight. Extensor digitorum longus (EDL) muscle weight and tail and tibia length were unchanged. In control rats, hGH induced an increase in all weight and length parameters. K+ deficiency was associated with reduced serum insulin-like growth factor I (IGF-I) and serum insulin but unchanged total 3,5,3'-triiodothyronine (TT3) and total thyroxine. hGH treatment of the K(+)-deficient rats restored serum IGF-I, but not serum insulin, and decreased TT3. In saline-treated K(+)-deficient rats [3H]ouabain binding site concentration decreased by 44 and 39% in soleus and EDL muscle, respectively, as compared with the saline-treated controls. hGH had no effect on the [3H]ouabain binding site concentration in the K(+)-deficient group, but, in control rats, increases of 11 and 8% were observed in soleus and EDL muscle, respectively. When the increase in muscle weight was taken into account, this amounted to relative increases of 24 and 30%, respectively. Low circulating GH and IGF-I levels are not the sole explanation for the growth retardation in K+ deficiency. GH/IGF-I stimulate the synthesis of Na(+)-K+ pumps in rats with an otherwise normal hormonal status.
Atrial natriuretic peptide synthesis in atrial tumors of transgenic mice
Authors: Gardner D. G. , Camargo M. J. , Behringer R. R. , Brinster R. L. , Baxter J. D. , Atlas S. A. , Laragh J. H. , Deschepper C. F.
Published: December 2017

Transgenic mice harboring a chimeric gene linking mouse protamine 1 5'-flanking sequence to the coding sequence of the simian virus 40 T-antigen develop spontaneous rhabdomyosarcomas of the right atria. The presence of the tumors is accompanied by dramatic elevations in plasma atrial natriuretic peptide (ANP) immunoreactivity (1,698 +/- 993 vs. 60 +/- 18 fmol/ml for controls) and hematocrit (56 +/- 8 vs. 51 +/- 2 for controls). The immunoreactive ANP (irANP) present in the tumors is similar in size to irANP found in normal mouse atria. ANP mRNA transcripts present in the tumors also appear to be very similar in overall size and 5'-termini to those produced in normal cardiac tissue. Microscopically, the tumors are composed of a disorganized array of densely packed abnormal-appearing cells. Immunocytochemistry and in situ hybridization analysis reveal considerable heterogeneity in ANP gene expression. ANP peptide and mRNA are detectable throughout the parenchyma of the tumors, but absolute levels of expression vary widely among different cells in the population. These tumors represent a potentially valuable model for the study of inappropriate ANP secretion and may provide a tissue source for the development of an ANP-producing atrial cell line.
Hepatic glucagon sensitivity and fasting glucose concentration in normal dogs
Authors: Bradley D. C. , Bergman R. N.
Published: December 2017

We assessed hepatic glucagon sensitivity in overnight-fasted, conscious dogs. Six pancreatic replacement protocols were performed in each of five animals. Somatostatin was infused to inhibit endogenous insulin and glucagon, insulin was replaced intraportally at 200 microU.min-1.kg-1, and glucagon was infused intraportally at 0, 0.6, 1, 2, 5, or 20 ng.min-1.kg-1. One intravenous glucose tolerance test was also performed in each animal for measurement of insulin sensitivity (SI). During hormone replacement at a given glucagon dose, plasma glucose differed substantially among animals (P = 0.003). Therefore the dose required for restoration of euglycemia (“glucagon requirement”) varied nearly sevenfold among animals, suggesting appreciable differences in glucagon sensitivity (GS). The latter was quantitated in individual animals as the initial slope of integrated glucose output vs. glucagon concentration. GS varied from 0.22 to 3.9 mg.kg-1.pg-1.ml among various animals and was inversely and significantly related to glucagon requirement. SI varied less (approximately 4-fold) and was not associated with glucagon requirement. These observations suggested that interanimal differences in glucose during hormone replacement were the result of substantial differences in GS. In addition, we found the GS of a given animal to be highly associated (P = 0.01) with its fasting glucose level. We conclude that GS varies substantially, and as such may be an important determinant of the fasting glucose level in normal animals.
Pyruvate carboxylase in genetic obesity
Authors: Lynch C. J. , McCall K. M. , Billingsley M. L. , Bohlen L. M. , Hreniuk S. P. , Martin L. F. , Witters L. A. , Vannucci S. J.
Published: December 2017

Immunoblotting and protein microsequencing were used to identify several adipocyte proteins expressed in an obesity-related fashion in the Zucker rat. One of these was a 116-kDa particulate protein (p116). The p116 levels in adipocytes from 5- to 7-wk-old obese Zucker rats were two- to fivefold higher on a per milligram of protein basis than levels in lean animals and decreased after the induction of streptozotocin-induced diabetes mellitus. This suggests the change may be related to the actions of insulin. Hepatic levels of p116 did not change. The p116 was purified to homogeneity from obese Zucker rat adipocytes, and polyclonal antisera were prepared against the purified protein in rabbits. Microanalysis of electroblotted p116 proteolytic fragments suggested that p116 was pyruvate carboxylase (PC). Other evidence that p116 was PC included the following: 1) p116 contained biotin, 2) p116 in particulate subcellular fractions was soluble after freeze-lysis, 3) antibodies to p116 reacted with purified hepatic PC, 4) p116 and purified hepatic PC had identical pI and relative molecular weight values, and 5) similar changes were detected in adipocyte p116 and PC enzyme activity during obesity and after the induction of streptozotocin-induced diabetes mellitus. Increased adipose tissue PC probably contributes to the increased lipogenic capacity of young obese Zucker rat adipocytes.
Peripheral and central angiotensin II regulates expression of genes of the renin-angiotensin system
Authors: Kohara K. , Brosnihan K. B. , Ferrario C. M. , Milsted A.
Published: December 2017

We investigated whether angiotensin (ANG) II has the potential to regulate expression of genes of the renin-angiotensin system (RAS) in peripheral and central tissues. ANG II (0.1 or 6.0 nmol/h) was infused by osmotic minipump into male Sprague-Dawley rats (225-250 g) for 5 days, either intravenously or intracerebroventricularly. We measured angiotensinogen mRNA in liver, adrenal glands, and brain (hypothalamus and lower brain stem), renin mRNA in the kidney, and angiotensin-converting enzyme (ACE) mRNA in the lung and testis by Northern blot analysis. We demonstrated that plasma ANG II increases the levels of liver angiotensinogen mRNA, decreases kidney renin mRNA, and decreases lung ACE mRNA. Intracerebroventricular administration of ANG II resulted in a different pattern of responses of the peripheral RAS components. Liver angiotensinogen mRNA was increased, and kidney renin mRNA was decreased by both doses of ANG II, whereas lung ACE mRNA remained unresponsive at either dose. Centrally mediated influences of ANG II are most likely indirect since plasma ANG II concentration was not changed. This study has revealed that ANG II has profound diverse effects that influence the regulation of its formation. Further, results indicate that genes of the RAS responded to exogenous ANG II in both tissue- and route-specific ways.
Liver releases galanin during sympathetic nerve stimulation
Authors: Kowalyk S. , Veith R. , Boyle M. , Taborsky G. J.
Published: December 2017

To determine whether the gut or liver releases galanin during sympathetic neural activation, we performed bilateral thoracic splanchnic nerve stimulation (BTSNS) in halothane-anesthetized dogs. Using experimentally determined galanin extraction rates of 60% for gut and no extraction by liver, calculations demonstrated a minor increase in gut spillover during BTSNS (delta = +4.8 +/- 1.8 pmol/min), whereas liver spillover of galanin-like immunoreactivity (GLIR) increased markedly (delta = +27.9 +/- 9.5 pmol/min). To confirm the finding of liver galanin release, GLIR was measured in femoral artery, portal vein, and hepatic vein during hepatic nerve stimulation (HNS). GLIR spillover from gut was not increased by HNS (delta = +1.9 +/- 6.3 pmol/min). In contrast, liver GLIR spillover was greatly increased during HNS (delta = +53.3 +/- 16.4 pmol/min). Extracts of canine liver contained 2.7 +/- 0.4 pmol GLIR/g tissue. We conclude that, despite the known significant galanin content of the gut, little galanin is released from this organ during sympathetic activation. In contrast, the liver, heretofore not described to contain galanin, contains and releases significant amounts of the peptide during sympathetic activation.
Sepsis-induced cascade of cytokine mRNA expression: correlation with metabolic changes
Authors: Byerley L. O. , Alcock N. W. , Starnes H. F.
Published: December 2017

The relationships between metabolic alterations and tissue-specific gene expression of tumor necrosis factor-alpha (TNF-alpha), interleukin 6 (IL-6), gamma-interferon (gamma-IFN), and interleukin 1 and serum levels of TNF-alpha and IL-6 before and after a live Escherichia coli septic challenge to rats were examined. From 0 to 2 h, serum glucose significantly decreased while plasma glucagon increased. By 8 h, plasma glucagon, serum insulin, and glucose appearance were significantly elevated. Gene expression of phosphoenolpyruvate carboxykinase increased 1 h after E. coli but by 4 h was significantly decreased. TNF-alpha mRNA (liver and spleen) and serum peptide levels peaked 1-2 h after the septic challenge and then decreased substantially by 6-8 h. Spleen IL-6 and gamma-IFN mRNA expression reached a maximum 4 h after E. coli challenge, whereas serum IL-6 levels were elevated by 2 h after injection of the bacteria. The increase in TNF-alpha mRNA and serum peptide levels correlated with the early fall in serum glucose and rise in plasma glucagon. Alterations in the rate of glucose appearance and plasma glucagon were observed later and coincided with the increased mRNA expression of IL-6 and gamma-IFN. Thus the metabolic alterations observed in the septic rat are associated with a complex cascade of several cytokines.
Theory of production rate calculations in steady and non-steady states and its application to glucose metabolism
Authors: Jacquez J. A.
Published: December 2017

I present a review and synthesis of the basic theory, steady state, and non-steady state for the calculation of metabolite production rates for systems that have a central well-mixed compartment that is the site of tracer input and sampling. The theory is then applied to the calculation of glucose production. If the only inputs are into the central compartment, an experimental design that involves varying tracer infusion rates to maintain constant specific activity in the central compartment and the same constant specific activity in the peripheral compartments allows calculation of the endogenous production. That holds even if the models are unidentifiable. The correct equation and Steele's pool fraction approximation reduce to the same result for this experimental design. However, that does not justify the use of Steele's equation when there are deviations from the exact experimental design. When the specific activity in the central compartment is not constant, model-dependent correction terms to Steele's equation are needed.
Effects of short-term testosterone exposure on insulin sensitivity of muscles in female rats
Authors: Holmang A. , Larsson B. M. , Brzezinska Z. , Bjorntorp P.
Published: December 2017

The effects of testosterone on insulin sensitivity were studied in oophorectomized female rats. Euglycemic, hyperinsulinemic clamp measurement showed a marked decrease of insulin sensitivity after 48 but not 24 h of testosterone exposure, which was overcome at high insulin concentrations. Insulin stimulation of 2-deoxyglucose uptake as well as glycogen synthesis was measured in the white and red parts of the gastrocnemius, the extensor digitorum longus, and soleus muscles as well as in the liver (only glycogen synthesis). After 24 h of treatment, inhibition of both 2-deoxyglucose uptake and glycogen synthesis was found in the most insulin-sensitive muscles. After 48 h of insulin stimulation, glycogen synthesis was inhibited in all examined individual muscles (white and red parts of gastrocnemius, extensor digitorum longus, and soleus) as was the activity of the insulin-sensitive part of glycogen synthase in muscle. Inhibition of insulin-stimulated 2-deoxyglucose uptake again affected the most insulin-sensitive muscles. There was a slight but significant change of muscle fiber composition toward less long-chain myosin and more short-chain myosin-containing fibers. Serum cortisol, plasma free fatty acids, and blood glycerol did not change. It is concluded that testosterone administration in moderate doses to oophorectomized female rats is followed by a rapid deterioration of insulin sensitivity in muscle, mediated mainly by perturbations of the insulin receptor-glycogen synthesis systems apparently coinciding with changes in muscle morphology.
Prednisone-induced osteopenia in beagles: variable effects mediated by differential suppression of bone formation
Authors: Quarles L. D.
Published: December 2017

To examine the mechanism of glucocorticoid-induced osteopenia and the basis for variable bone loss after glucocorticoid administration, we gave prednisone (1.3 mg.kg-1.day-1) to normal male dogs (n = 15) for 29 wk to attempt induction of osteopenia. Compared with age-matched control dogs (n = 14), prednisone treatment rapidly decreased spinal bone density by 4.3%, as assessed by quantitative digital radiography, and reduced trabecular bone volume by 14.6%, as measured by quantitative histomorphology of iliac crest bone specimens. Bone loss was attenuated in prednisone-treated dogs after prolonged treatment (greater than 12 wk). Prednisone treatment resulted in diminished bone formation rates (15 +/- 3.4 vs. 47 +/- 4.5 microns/yr) and activation frequency (0.4 +/- 0.1 vs. 1.3 +/- 0.2/day). These findings indicate that suppression of osteoblastic function and recruitment is the primary histological abnormality mediating glucocorticoid-induced osteopenia in beagles. In contrast, prednisone administration had no effect on bone resorption or serum concentrations of parathyroid hormone and 1,25-dihydroxyvitamin D, which suggests that these factors are not essential for prednisone-induced bone loss. Moreover, 33% of beagles were totally resistant to glucocorticoid-induced osteopenia. Such heterogeneity of bone loss was associated with variable suppressive effects of prednisone on osteoblastic function, as evidenced by greater bone formation rates and activation frequency in prednisone-resistant animals. Collectively, these observations suggest that glucocorticoid-induced bone loss results from a dynamic interplay between steroid-mediated suppression of osteoblastic function and recruitment and undefined compensatory factors that ameliorate the effects of glucocorticoids on osteoblastic precursors.
Use of 14CO2 in estimating rates of hepatic gluconeogenesis
Authors: Esenmo E. , Chandramouli V. , Schumann W. C. , Kumaran K. , Wahren J. , Landau B. R.
Published: December 2017

Estimating the rate of hepatic gluconeogenesis in vivo from the incorporation of 14C from 14CO2 into glucose requires determination of the rates in liver of equilibration of oxaloacetate with fumarate, conversion of oxaloacetate to phosphoenolpyruvate (PEP), and conversion of PEP to pyruvate, all relative to the rate of tricarboxylic acid cycle flux. With the use of a model of mitochondrial metabolism and gluconeogenesis, expressions are derived relating specific activity of carboxyl of PEP from 14CO2 to those rates and specific activity of mitochondrial CO2. If those rates and specific activity of mitochondrial CO2 are known, specific activity of PEP, calculated using the expressions, should, on a mole basis, be one-half the specific activity of the glucose formed. At steady state, in the 60-h fasted individual, where glucose formation is solely by gluconeogenesis, twice estimated specific activity of PEP should then approximate that of blood glucose. Estimates of relative rates in 60-h fasted humans, previously made from distribution of 14C in glutamate from phenylacetylglutamine excreted when [3-14C]lactate and phenylacetate were given, were applied to the expressions. Specific activity of mitochondrial CO2 was equated to that of CO2 expired by 60-h fasted subjects given NaH14CO3 and alpha-[1-14C]ketoisocaproate. Predicted specific activities approximated actual specific activities of blood glucose when NaH14CO3 was administered. alpha-[1-14C]ketoisocaproate administrations gave underestimates. This is attributable to differences between specific activities of hepatic mitochondrial CO2 and expired CO2, which is evidenced by higher incorporations of 14C in glucose than in expired CO2 from alpha-[1-14C]ketoisocaproate than from NaH14CO3.(ABSTRACT TRUNCATED AT 250 WORDS
Mechanism of delayed hepatic glycogen synthesis after an oral galactose load vs. an oral glucose load in adult rats
Authors: Niewoehner C. B. , Neil B.
Published: December 2017

We have compared the effects of administration of oral galactose or glucose (1 g/kg) to 24-h fasted rats to examine the mechanism by which galactose regulates its own incorporation into liver glycogen in vivo. Liver glycogen increased to a maximum more slowly after galactose than after glucose administration (0.14 vs. 0.29 mumol.g liver-1.min-1). Glycogen accumulation after the galactose load was 70% of that after the glucose load (149 vs. 214 mumol), and the net increase in liver glycogen represented the same proportion (24 vs. 22%) of added carbohydrate after urinary loss of galactose was accounted for. Slower glycogen accumulation after galactose vs. glucose loading could not be explained by galactosuria, by differences in the active forms of synthase or phosphorylase, by end product (glycogen) inhibition of synthase phosphatase, or by different concentrations of the known allosteric effectors of synthase R plus I and phosphorylase a. Similar increases in glucose 6-phosphate were observed after both hexoses. AMP and ADP increased only transiently after galactose administration, and ATP, UTP, and Pi concentrations were unchanged. The UDP-glucose concentration decreased, whereas the UDP-galactose concentration increased two- to threefold after galactose but not glucose administration. The UDP-glucose pyrophosphorylase reaction is inhibited competitively by UDP-galactose. This could explain the decreased UDP-glucose concentration and the reduced rate of glycogen synthesis after galactose was given.
PDGF induces tyrosine phosphorylation in osteoblast-like cells: relevance to mitogenesis
Authors: Davidai G. , Lee A. , Schvartz I. , Hazum E.
Published: December 2017

We have studied the association between protein tyrosine phosphorylation and the mitogenic effect induced by platelet-derived growth factor (PDGF) in the osteoblast-like cell line MC3T3-E1. PDGF caused a dose-dependent increase in [3H]thymidine incorporation in MC3T3-E1 cells, reaching a plateau at 10 ng/ml. Vanadate, a potent phosphatase inhibitor, induced a twofold increase in thymidine incorporation. The combination of vanadate and PDGF resulted in a dose-dependent synergistic effect on thymidine incorporation. Genistein, a tyrosine kinase inhibitor, inhibited in a dose-related manner (2-20 microM) the mitogenic effect induced by either PDGF or the combination of vanadate and PDGF. These observations suggest that tyrosine kinases are involved in mediating the mitogenic effect of PDGF in these cells. PDGF treatment of MC3T3-E1 cells and subsequent immunoblotting with antiphosphotyrosine antibodies resulted in a marked phosphorylation of the PDGF receptor. Vanadate had a lesser effect on PDGF receptor phosphorylation, but given together with PDGF it induced a significant increase in the intensity of receptor phosphorylation. Preincubation with genistein abrogated these effects. Taken together, these findings indicate a direct correlation between thymidine incorporation and tyrosine phosphorylation in MC3T3-E1 cells and suggest that tyrosine phosphorylation plays a role in PDGF-induced mitogenic activity in osteoblast-like cells.
Effects of prior exercise on the action of insulin-like growth factor I in skeletal muscle
Authors: Henriksen E. J. , Louters L. L. , Stump C. S. , Tipton C. M.
Published: December 2017

Prior exercise increases insulin sensitivity for glucose and system A neutral amino acid transport activities in skeletal muscle. Insulin-like growth factor I (IGF-I) also activates these transport processes in resting muscle. It is not known, however, whether prior exercise increases IGF-I action in muscle. Therefore we determined the effect of a single exhausting bout of swim exercise on IGF-I-stimulated glucose transport activity [assessed by 2-deoxy-D-glucose (2-DG) uptake] and system A activity [assessed by alpha-(methylamino)isobutyric acid (MeAIB) uptake] in the isolated rat epitrochlearis muscle. When measured 3.5 h after exercise, the responses to a submaximal concentration (0.2 nM), but not a maximal concentration (13.3 nM), of insulin for activation of 2-DG uptake and MeAIB uptake were enhanced. In contrast, prior exercise increased markedly both the submaximal (5 nM) and maximal (20 nM) responses to IGF-I for activation of 2-DG uptake, whereas only the submaximal response to IGF-I (3 nM) for MeAIB uptake was enhanced after exercise. We conclude that 1) prior exercise significantly enhances the response to a submaximal concentration of IGF-I for activation of the glucose transport and system A neutral amino acid transport systems in skeletal muscle and 2) the enhanced maximal response for IGF-I action after exercise is restricted to the signaling pathway for activation of the glucose transport system.
Increased alpha 1-adrenoceptor density in brown adipose tissue indicates recruitment drive in hypothyroid rats
Authors: Dicker A. , Raasmaja A. , Cannon B. , Nedergaard J.
Published: December 2017

The effects of hypothyroidism on whole body thermogenesis, brown adipose tissue recruitment state, and alpha 1-adrenergic receptor density were investigated. Treatment of rats with methimazole for 4-5 wk led, as expected, to reduction of growth and resting metabolic rate. The thermogenic response to norepinephrine injection was practically abolished. Generally, only small effects of hypothyroidism on brown adipose tissue were observed: total protein content, mitochondrial GDP binding capacity, and total content of the uncoupling protein thermogenin were not altered. The density of beta-adrenergic receptors (estimated with [3H]CGP-12177 as a ligand) was also unchanged. However, the density of alpha 1-adrenergic receptors (estimated with [3H]prazosin) was markedly increased; in other physiological conditions, such an increase has been associated with an increased degree of recruitment of the tissue. These data indicate that brown adipose tissue in the subthermoneutral hypothyroid animal, probably due to homeostatic mechanisms, is exposed to an increased sympathetic stimulation, leading to an increased alpha 1-adrenoceptor density. However, other features of recruitment are only poorly induced, probably due to attenuation of the beta-adrenergic signaling mechanism. The increased alpha 1-adrenergic receptor density may be responsible for certain altered features of brown adipose tissue in hypothyroid animals, such as peroxisomal recruitment and perhaps also for maintenance of the thermogenin content. The results also indicate that the increased alpha 1-adrenergic density generally seen in recruitment would not result from chronic beta-adrenergic stimulation of the tissue but may be controlled via another regulatory pathway, e.g., via the alpha 1-adrenergic pathway itself.
Effect of hyperinsulinemia on ovine fetal leucine kinetics during prolonged maternal fasting
Authors: Liechty E. A. , Boyle D. W. , Moorehead H. , Liu Y. M. , Denne S. C.
Published: December 2017

The primary effect of insulin on whole body protein metabolism in postnatal life is to reduce proteolysis. To assess the role of insulin in the regulation of protein metabolism in prenatal life, leucine kinetics were determined in the ovine fetus at baseline and in response to hyperinsulinemia. These measurements were made in each fetus in two different maternal states: ad libitum maternal feeding and after a 5-day maternal fast. Maternal fasting resulted in significant increases in baseline fetal leucine rate of appearance (Ra; 51.9 +/- 16.7 vs. 37.3 +/- 3.6 mumol/min, P < 0.05) and leucine oxidation (30.1 +/- 8.9 vs. 8.8 +/- 2.2 mumol/min, P < 0.05). Hyperinsulinemia, which was associated with significant increases in fetal glucose utilization, did not affect total fetal leucine R(a) or leucine release from fetal proteolysis in either maternal state. Under well-fed maternal conditions, hyperinsulinemia produced no changes in the fetal oxidative or nonoxidative disposal of leucine. In contrast, during maternal fasting, hyperinsulinemia reduced fetal leucine oxidation (11.0 +/- 3.7 vs. 31.1 +/- 8.9 mumol/min, P < 0.05) and increased the nonoxidative disposal of leucine (35.4 +/- 4.0 vs. 19.0 +/- 6.1 mumol/min, P < 0.05). This resulted in a change in the fetal leucine accretion rate from negative to positive (-20.9 +/- 7.5 vs. 7.5 +/- 6.7 mumol/min, P < 0.05). These results suggest that, under conditions of restricted maternal substrate intake, fetal hyperinsulinemia and the attendant increase in fetal glucose utilization are associated with increased protein synthesis rather than decreased protein breakdown, thereby improving fetal leucine carcass accretion.
Sites of infusion and sampling for measurement of rates of production in steady state
Authors: Norwich K. H.
Published: December 2017

When labeled and unlabeled (endogenously produced) substances enter an organism through different ports of entry, the anatomical location of these entry sites must be taken into account explicitly in calculating rates of appearance. All traditional formulas that are used for calculating rates of appearance are based on the assumption that labeled and unlabeled substances enter by means of the same port. These formulas are, therefore, not in general valid for metabolites such as lactate, where the entry ports differ. In such cases, specific activity will not be uniform throughout the organism even when the labeled and unlabeled substances are both in steady state. One cannot speak of the (unique) specific activity, because none exists. It is useful in these cases to define a distributed specific activity, which is the ratio of the concentration of labeled substance at one anatomical site to the concentration of unlabeled substance at another. We can then show that the rate of appearance for the double steady state (steady tracer infusion method) is given, approximately, as the ratio of the rate of infusion of the labeled substance to a particular distributed specific activity.
Adrenergic regulation of type II 5'-deiodinase circadian rhythm in rat harderian gland
Authors: Osuna C. , Jimenez J. , Reiter R. J. , Rubio A. , Guerrero J. M.
Published: December 2017

This paper reports on the regulation of the nyctohemeral profile of type II thyroxine 5'-deiodinase (T45'D) activity in the rat harderian gland. Harderian gland T45'D activity exhibits a nighttime increase with maximal values late in the dark period (0200–0400 h) and basal values during the daytime. The nocturnal rise of the deiodinating activity was prevented by either exposure of animals to light at night, injecting the animals with both alpha- and beta-adrenergic receptor blockers, or bilateral superior cervical ganglionectomy (SCGx). However, adrenalectomy did not affet the enzyme activity in the harderian gland. In brown adipose tissue (BAT), where thyroid hormone metabolism is extremely dependent on alpha 1-adrenergic stimulation by blood-circulating catecholamines, adrenalectomy significantly decreased T45'D activity. Deiodinating activities in brain frontal cortex (BFC) and pituitary gland were unaffected by adrenalectomy. Unlike in the harderian gland, SCGx did not modify the T45'D activity in either BAT, BFC, or the pituitary gland. The results suggest that elevated plasma catecholamines are not required for harderian gland T45'D activation and that the nyctohemeral profile of the enzyme activity in the harderian gland is dependent on the noradrenergic input from the superior cervical ganglia.
Complex effects of arachidonic acid and its lipoxygenase products on cytosolic calcium in GH3 cells
Authors: Vacher P. , McKenzie J. , Dufy B.
Published: December 2017

The mechanisms by which arachidonic acid (AA) and its metabolites stimulate prolactin release from pituitary cells are not understood. Because Ca2+ ions are pivotal to exocytosis, we investigated the effects of AA metabolites on intracellular calcium concentration ([Ca2+]i) and membrane ionic currents using dual-excitation microspectrofluorimetry and whole cell patch-clamp techniques in GH3/B6 pituitary line cells. AA (1 microM) had a biphasic effect on [Ca2+]i, mobilization of intracellular Ca2+, followed by stimulation of Ca2+ entry. Only the latter appeared to result from the degradation of AA through the lipoxygenase pathway. Indomethacin (Indo, 10 microM) and lipoxygenase products 5-, 12-, and 15-hydroxyeicosatetraenoic acid (HETE, 1 microM), increased action potential duration (12-HETE) or frequency (5- and 15-HETE, Indo). These effects depended on inhibition of d-tubocurarine- or tetraethylammonium-sensitive K+ conductances and stimulation of voltage-dependent Ca2+ channels. Refilling of intracellular Ca2+ stores, and Ca2+ efflux, may also be stimulated. Our results demonstrate a control of [Ca2+]i by a second messenger (AA) and its metabolites (HETEs).
Visceral fat accumulation in obese subjects: relation to energy expenditure and response to weight loss
Authors: Leenen R. , van der Kooy K. , Deurenberg P. , Seidell J. C. , Weststrate J. A. , Schouten F. J. , Hautvast J. G.
Published: December 2017

Seventy-eight healthy obese subjects, 40 premenopausal women and 38 men aged 27-51 yr received a 4.2 MJ/day energy-deficit diet for 13 wk. Resting metabolic rate (RMR) and diet-induced thermogenesis (DIT) were measured by indirect calorimetry. Abdominal subcutaneous and visceral fat areas were calculated from magnetic resonance imaging scans before and after weight loss. Before weight loss, visceral fat accumulation was positively correlated with higher levels of RMR (P < 0.05) and DIT (P < 0.01) in women but not in men. The mean weight reduction was 12.2 +/- 3.5 (SD) kg. In men but not in women, an initially large visceral fat depot was associated with a reduced loss of weight and total fat mass (P < 0.05). Within each sex, an initial abundance of visceral fat was significantly related to a larger loss of visceral fat (P < 0.001) and in men to a smaller loss of subcutaneous fat (P < 0.05). These results suggest that there may be gender differences in the associations between visceral fat accumulation and components of energy expenditure (RMR and DIT) in obese subjects. Obese subjects with an initial abundance of visceral fat do not lose more body weight but more visceral fat than subjects with less visceral fat.
Precision and accuracy of doubly labeled water energy expenditure by multipoint and two-point methods
Authors: Cole T. J. , Coward W. A.
Published: December 2017

Two-point or multipoint, that is the question. Equations are developed to compare the precision and accuracy of energy expenditure, as estimated from doubly labeled water data, when analyzed by the multipoint and two-point methods. The equations convert the enrichments of deuterium and oxygen-18 into their ratio and product, quantities that are less covariant than the two isotopes themselves are. This is important not only for estimating the precision but also as a graphical aid, since the ratios of the enrichments model carbon dioxide production, whereas the enrichment products largely model water turnover. Using data on 12 human subjects from the United Kingdom and The Gambia as examples, the combined precision and accuracy of the multipoint method (CV 3.6%) was found to be appreciably better than the two-point method (CV 5.4%). The bias in the multipoint estimate of body pool size would need to be three times as large as was observed before it canceled out the better precision.
Multiple metabolic effects of CGRP in conscious rats: role of glycogen synthase and phosphorylase
Authors: Rossetti L. , Farrace S. , Choi S. B. , Giaccari A. , Sloan L. , Frontoni S. , Katz M. S.
Published: December 2017

Calcitonin gene-related peptide (CGRP) is a neuropeptide that is released at the neuromuscular junction in response to nerve excitation. To examine the relationship between plasma CGRP concentration and intracellular glucose metabolism in conscious rats, we performed insulin (22 pmol.kg-1.min-1) clamp studies combined with the infusion of 0, 20, 50, 100, 200, and 500 pmol.kg-1.min-1 CGRP (plasma concentrations ranging from 2 x 10(-11) to 5 x 10(-9) M). CGRP antagonized insulin's suppression of hepatic glucose production at plasma concentrations (approximately 10(-10) M) that are only two- to fivefold its basal portal concentration. Insulin-mediated glucose disposal was decreased by 20-32% when CGRP was infused at 50 pmol.kg-1.min-1 (plasma concentration 3 x 10(-10) M) or more. The impairment in insulin-stimulated glycogen synthesis in skeletal muscle accounted for all of the CGRP-induced decrease in glucose disposal, while whole body glycolysis was increased despite the reduction in total glucose uptake. The muscle glucose 6-phosphate concentration progressively increased during the CGRP infusions. CGRP inhibited insulin-stimulated glycogen synthase in skeletal muscle with a 50% effective dose of 1.9 +/- 0.36 x 10(-10) M. This effect on glycogen synthase was due to a reduction in enzyme affinity for UDP-glucose, with no changes in the maximal velocity. In vitro CGRP stimulated both hepatic and skeletal muscle adenylate cyclase in a dose-dependent manner. These data suggest that 1) CGRP is a potent antagonist of insulin at the level of muscle glycogen synthesis and hepatic glucose production; 2) inhibition of glycogen synthase is its major biochemical action in skeletal muscle; and 3) these effects are present at concentrations of the peptide that may be in the physiological range for portal vein and skeletal muscle. These data underscore the potential role of CGRP in the physiological modulation of intracellular glucose metabolism.
Effects of chronic elevation in plasma cortisol on hepatic carbohydrate metabolism
Authors: Goldstein R. E. , Wasserman D. H. , McGuinness O. P. , Lacy D. B. , Cherrington A. D. , Abumrad N. N.
Published: December 2017

This study was undertaken to investigate the effects of chronic physiological elevations in plasma cortisol on glycogenolysis and gluconeogenesis in conscious, overnight-fasted dogs. Experiments consisted of an 80-min tracer and dye equilibration period and a 40-min sampling period. Infusions of D-[3-3H]glucose, L-[U-14C]alanine, and indocyanine green dye were used to assess glucose production (Ra) and gluconeogenesis using tracer and arteriovenous (a-v) difference techniques. In the cortisol group, (n = 10), a continuous infusion of hydrocortisone (3.5 micrograms.kg-1 x min-1) was begun 5 days before the experiment and continued throughout the sampling period. In the saline group (n = 10), there was no infusion of cortisol. The fivefold elevation in plasma cortisol increased plasma insulin from 12 +/- 2 to 19 +/- 2 microU/ml. Glucose Ra was elevated in the cortisol group (3.5 +/- 0.2 vs. 2.8 +/- 0.2 mg.kg-1 x min-1) but net hepatic glucose output was markedly diminished (1.2 +/- 0.4 vs. 2.7 +/- 0.3 mg.kg-1 x min-1). Gluconeogenic conversion of alanine to glucose was increased slightly by cortisol (0.60 +/- 0.13 to 0.99 +/- 0.12 mumol.kg-1 x min-1), but the gluconeogenic efficiency of the liver was unchanged. Cortisol increased hepatic glycogen content evident at the end of the study greater than twofold (76.4 +/- 7.9 vs. 30.0 +/- 4.7 g/liver). These results suggest that cortisol 1) promotes glucose cycling through glycogen, 2) greatly inhibits nonhepatic glucose utilization, 3) increases hepatic gluconeogenesis in vivo primarily through enhanced substrate delivery to the liver, and 4) raises plasma insulin levels, which restrains intrahepatic gluconeogenesis.
Estimation of protein fractional synthetic rate from tracer data
Authors: Toffolo G. , Foster D. M. , Cobelli C.
Published: December 2017

The fractional synthetic rate (FSR) is a key parameter characterizing protein turnover that is estimated from tracer kinetic data. Formulas to estimate this parameter usually assume a precursor-product model. Assuming this model is correct, we discuss these formulas to estimate the FSR in the steady and non-steady state both for the radioactive and stable isotope tracer. Then we deal with the non-steady-state case where the FSR becomes time varying and derive formulas for its estimation. A non-steady-state case study on the flooding-dose technique for measuring protein turnover is presented.
Insulin resistance in polycystic ovary syndrome: decreased expression of GLUT-4 glucose transporters in adipocytes
Authors: Rosenbaum D. , Haber R. S. , Dunaif A.
Published: December 2017

We have found that women with polycystic ovary syndrome (PCOS) have decreased sensitivity and responsiveness to insulin. The present study was performed to determine whether this impaired insulin responsiveness was associated with diminished GLUT-4 glucose transporter content in adipocytes. Insulin-stimulated glucose transport and GLUT-4 abundance were measured in abdominal adipocytes from obese (n = 9) and lean (n = 7) PCOS as well as obese (n = 8) and lean (n = 8) control women matched for age and weight. No woman had impaired glucose tolerance. The maximal insulin-stimulated increment in adipocyte glucose transport was independently decreased by obesity and by PCOS. As expected, GLUT-4 content in adipocyte membranes was decreased in obesity (by 40%, P < or = 0.01). GLUT-4 content was also significantly decreased in PCOS (by 36%, P < or = 0.01), independent of obesity. There was a highly significant correlation (R = 0.66, P < = 0.001) between GLUT-4 content and insulin-stimulated glucose transport in adipocytes from individual women across the study population. We conclude that the diminished adipocyte insulin responsiveness in PCOS is associated with decreased GLUT-4 abundance. This represents a newly recognized phenotypic feature of the insulin resistance of PCOS. Moreover, in human adipocytes, GLUT-4 abundance is highly correlated with insulin responsiveness.