bluuebunny/crossref_metadata_2025 huggingface dataset:
Lipid transport function of lipoproteins in blood plasma
Authors: Havel R. J.
Published: December 2017

Fatty acid and cholesterol transport in plasma lipoproteins evolved in the context of an open circulatory system in which lipoprotein particles are secreted directly into the blood and have ready access to cells in various tissues. In higher vertebrates with closed capillary beds, hydrolysis of triglycerides at capillary surfaces is required for efficient uptake of their component fatty acids into cells. Likewise, hydrolysis of cellular triglycerides in cells of adipose tissue precedes mobilization of the fatty acids and permits large amounts to be transported in the blood. However, in all Metazoa lipoproteins are secreted primarily from cells adjacent to an open microvascular bed. Uptake of lipoprotein particles as such into cells occurs in invertebrates and vertebrates alike, facilitated by binding to high-affinity receptors on cell surfaces. In vertebrates, a concentration gradient created between cholesterol in cells and lipoproteins by a cholesterol-esterifying enzyme that acts on lipoproteins promotes movement of cholesterol into the plasma compartment. Thus the strategies to transport poorly soluble lipids include enzymatic reactions at cell surfaces and in blood plasma as well as the processes of exocytosis and endocytosis.
Parathyroid hormone acutely elevates intracellular calcium in osteoblastlike cells
Authors: Reid I. R. , Civitelli R. , Halstead L. R. , Avioli L. V. , Hruska K. A.
Published: December 2017

Changes in cytoplasmic calcium concentration ([Ca2+]i) activate numerous cellular processes thus mediating the effects of a number of hormones, but whether this mechanism is involved in the activation of osteoblasts by parathyroid hormone (PTH) remains uncertain. To examine this question, [Ca2+]i has been measured in suspensions of UMR 106 cells, a rodent osteosarcoma cell line with an osteoblastic phenotype. Basal [Ca2+]i was 137 +/- 3.7 nM (n = 60) and after the addition of rat PTH-(1–34) [rPTH-(1-34)] there was a rapid, dose-related increase with return to base line within 1 min. Half-maximal stimulation was produced by 5 X 10(-8) M rPTH-(1-34). Complexing of intracellular calcium by EGTA addition immediately before that of rPTH did not affect the calcium transient; neither did MnCl2 (10(-4) M) nor diltiazem (10(-4) M). Verapamil (10(-5) M) reduced the [Ca2+]i peak height after rPTH to 0.48 +/- 0.14 of control (n = 7). 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoic acid and dantrolene both reduced the [Ca2+]i response to rPTH (0.65 +/- 0.08 and 0.29 +/- 0.13 of control, respectively). Forskolin (10(-6) and 10(-5) M) produced a slight [Ca2+]i transient smaller in amplitude than seen with PTH. It is concluded that PTH mobilizes an intracellular calcium pool in these osteoblastlike cells, and the predominant mechanism for this is independent of cAMP.
1,25-Dihydroxyvitamin D3 target cells in immature pancreatic islets
Authors: Clark S. A. , Stumpf W. E. , Sar M. , DeLuca H. F.
Published: December 2017

Target cells of 1,25-dihydroxyvitamin D3 were identified by autoradiography in islets from rats of different ages. Nuclei of pancreatic islet cells selectively concentrated 1,25-[3H]dihydroxyvitamin D3 but not 25-[3H]hydroxyvitamin D3 or 24,25-[3H]dihydroxyvitamin D3. Developmental studies of pancreatic islets indicated that target cells, as revealed by significant nuclear concentration of 1,25-[3H]dihydroxyvitamin D3, are present in islet cells of fetal rats. The percentage of islet cells that concentrated 1,25-[3H]dihydroxyvitamin D3 increased from 10 to 15% in the fetus to 60% at 1 day of age. Immunocytochemical staining indicated that insulin-containing cells but not glucagon or somatostatin cells concentrated 1,25-[3H]dihydroxyvitamin D3. Peak uptake of 1,25-[3H]dihydroxyvitamin D3 was calculated to be 400 pmol/mg DNA, with no significant difference in nuclear accumulation between islet cells from neonatal and adult rats or between islets in vivo and isolated islets in vitro. The results of these studies indicate that 1,25-[3H]dihydroxyvitamin D3 target cells are present in islets before pancreatic beta-cells are morphologically or functionally mature; islet beta-cells concentrate 1,25-dihydroxyvitamin D3, but not 25-hydroxyvitamin D3 or 24,25-dihydroxyvitamin D3. We conclude that only the 1,25-dihydroxyvitamin D3 metabolite of vitamin D is accumulated by nuclei of developing and mature beta-cells and suggest that 1,25-dihydroxyvitamin D3 plays a role in the maturation of islet beta-cells.
Energy expenditure after infusion of glucose-based total parenteral nutrition
Authors: Gil K. M. , Askanazi J. , Elwyn D. H. , Gump F. E. , Kinney J. M.
Published: December 2017

Resting energy expenditure (REE), carbohydrate balance, and lipogenesis were calculated after administration of glucose-based total parenteral nutrition (TPN) to determine whether the thermic effect of glucose is equal to the energy cost of storing the glucose. Estimated cost of storage as glycogen (5.3%) and fat (19%) was compared with measured increases in REE. Patients with malnutrition received 5% dextrose in water and 6 days of TPN with a low (1.2 times REE, group 1) or high (2.0 times REE, group 2) level of glucose intake. Increases in REE by day 6 were 10% (group 1) and 28% (group 2). The theoretical cost of glucose storage as glycogen and fat accounted for approximately 40% of the measured increase in REE in patients in group 2. The thermic effect of TPN (derived from patients in group 1) accounted for most of the balance. The majority of the thermic effect of high levels of glucose infused with TPN can be explained on the basis of the thermic effect of TPN and glucose storage.
Cold exposure potentiates the effect of insulin on in vivo glucose uptake
Authors: Vallerand A. L. , Perusse F. , Bukowiecki L. J.
Published: December 2017

The effects of cold exposure (48 h at 4 degrees C) and insulin injection (0.5 U/kg iv) on the rates of net 2-[3H]deoxyglucose uptake (Ki) in peripheral tissues were investigated in warm-acclimated rats (25 degrees C). Cold exposure and insulin treatment independently increased Ki values in skeletal muscles (soleus, extensor digitorum longus, and vastus lateralis), heart, white adipose tissue (subcutaneous, gonadal, and retroperitoneal), and brown adipose tissue (P less than 0.01). The effects of cold exposure were particularly evident in brown adipose tissue where the Ki increased greater than 100 times. When the two treatments were combined (insulin injection in cold-exposed rats), it was found that cold exposure synergistically enhanced the maximal insulin responses for glucose uptake in brown adipose tissue, all white adipose tissue depots, and skeletal muscles investigated. The results indicate that cold exposure induces an "insulin-like" effect on Ki that does not appear to be specifically associated with shivering thermogenesis in skeletal muscles, because that effect was observed in all insulin-sensitive tissues. The data also demonstrate that cold exposure significantly potentiates the maximal insulin responses for glucose uptake in the same tissues. This potentialization may result from an enhanced responsiveness of peripheral tissues to insulin, possibly occurring at metabolic steps lying beyond the insulin receptor and an increased tissue blood flow augmenting glucose and insulin availability and thereby amplifying glucose uptake.
Evaluation of a parathyroid hormone antagonist in an in vivo multiparameter bioassay
Authors: Horiuchi N. , Rosenblatt M.
Published: December 2017

The antagonist properties of a bovine parathyroid hormone analogue ([Tyr34]bPTH-(7-34] amide were quantitatively assessed in vivo in a multiparameter assay to estimate the potency of the antagonist against the major actions of PTH. The analogue inhibited PTH-stimulated urinary excretion of phosphate and adenosine 3',5'-cyclic monophosphate in vitamin D-deficient thyroparathyroidectomized rats in a dose-dependent manner. At a molar dose ratio as low as 5:1 of antagonist to PTH, partial inhibition occurred. PTH stimulates the activity of 25-hydroxyvitamin D3-1 alpha-hydroxylase in renal proximal tubules. When coinfused with PTH, this analogue completely inhibited PTH-stimulated 1 alpha-hydroxylase activity at a molar dose ratio of 25:1 of antagonist to PTH and partially inhibited the activity at a molar dose ratio of 10:1. The analogue revealed no PTH-like agonist activity for stimulation of the 1 alpha-hydroxylase. Taken together, these studies indicate that [Tyr34]bPTH-(7-34) amide is a potent antagonist of several of the parameters of PTH action in vivo and demonstrate the feasibility of designing a PTH antagonist that can interact simultaneously with all the PTH receptors responsible for the hormone's major actions in vivo.
A primed-infusion technique for rapid estimation of the metabolic clearance rate of 1,25(OH)2D3
Authors: Eastell R. , Riggs B. L. , Kumar R.
Published: December 2017

We have developed a rapid primed-infusion technique for the measurement of the metabolic clearance and production rate of 1,25-dihydroxyvitamin D3 in normal human subjects and experimental animals. With this method, an estimate of the metabolic clearance rate of 1,25-dihydroxyvitamin D3 can generally be made within 3 to 4 h. Initial studies in five dogs using 1,25-[3H]-dihydroxyvitamin D3 (180 Ci/mmol) allowed us to determine the optimal ratio of loading dose to infusion rate that resulted in the most rapid attainment of steady-state levels of plasma radioactivity. By use of this technique we found that the metabolic clearance rate of 1,25-dihydroxyvitamin D3 in dogs was 6.3 +/- 1.2 ml/min (mean +/- SD); the production rate of the hormone was 0.40 +/- 0.25 microgram/day (20.4 +/- 14.4 ng . kg-1 . day-1). In eight normal women, aged 28-51 yr, the metabolic clearance rate for 1,25-dihydroxyvitamin D3 was 25.9 +/- 4.7 ml/min; the production rate was 1.38 +/- 0.45 microgram/day (20.7 ng . kg-1 . day-1). The advantages of this method relative to ones used in the past are that it can be performed quickly (generally within 3-4 h) with the use of only tracer amounts of this hormone (equivalent to 1.1% of the production rate). With this method, no assumptions about the most appropriate model to which to fit the data need to be made. Because of its rapidity, no metabolites of the injected 1,25-dihydroxyvitamin D3 are formed during the study interval.
Age-associated alterations in hepatic beta-adrenergic receptor/adenylate cyclase complex
Authors: Graham S. M. , Herring P. A. , Arinze I. J.
Published: December 2017

The effect of age on catecholamine regulation of hepatic glycogenolysis and on hepatic adenylate cyclase was studied in male rats up to 24 mo of age. Epinephrine and norepinephrine stimulated glycogenolysis in isolated hepatocytes at all age groups studied. Isoproterenol, however, stimulated glycogenolysis only at 24 mo. In isolated liver membranes, usual activators of adenylate cyclase increased the activity of the enzyme considerably more in membranes from 24-mo-old rats than in membranes from either 3- or 21-mo-old rats. The Mn2+-dependent activity of the cyclase was increased by 2.9-fold in 3-mo-old animals and approximately 5.7-fold in 24-mo-old rats, indicating a substantial age-dependent increase in the intrinsic activity of the catalytic unit. The density of the beta-adrenergic receptor, as measured by the binding of [125I]-iodocyanopindolol to plasma membranes, was 5-8 fmol/mg protein in rats aged 3-12 mo but increased to 19 fmol/mg protein in 24-mo-old rats. Computer-aided analysis of isoproterenol competition of the binding indicated a small age-dependent increase (from 30% at 3 mo to 43% at 24 mo) in the proportion of beta-receptors in the high-affinity state. These observations suggest that beta-receptor-mediated hepatic glycogenolysis in the aged rat is predicated upon increases in the density of beta-receptors as well as increased intrinsic activity of the catalytic unit of adenylate cyclase.
Absence of detectable phosphocreatine in rat luteal cells
Authors: Cross J. C. , Soodak L. K. , Musicki B. , Behrman H. R.
Published: December 2017

Previous studies from this laboratory showed that adenosine amplifies the action of luteinizing hormone (LH) severalfold in rat and human luteal cells by an intracellular, adenosine 5'-triphosphate (ATP)-linked process. The objective of this study was to evaluate the contribution of phosphocreatine (PCr) and creatine kinase (CK) to the dynamics of luteal ATP metabolism. Levels of PCr in luteinized rat ovaries were similar to those seen in liver but were approximately 1 and 7% of levels found in skeletal and heart muscle, respectively. In isolated rat luteal cells, little detectable PCr was seen after incubation in the presence or absence of adenosine, although cell ATP levels were increased twofold by adenosine treatment. The presence or absence of LH had no effect on either PCr or ATP levels in incubations of isolated luteal cells. Analysis of CK activity in tissue and cell homogenates showed that the specific activity of CK in luteal cells was in the same range as that seen in liver but less than 1/30 of that seen in skeletal muscle. From these studies we conclude that rat luteal cells contain little, if any, PCr and low levels of CK. Thus the rapid changes in ATP levels that are seen in rat luteal tissue and cells may occur because these cells have little capacity to buffer ATP levels with a reservoir of high-energy phosphate groups in the form of PCr.
Altered contractile proteins in skeletal muscle of diabetic rats
Authors: Ganguly P. K. , Taira Y. , Elimban V. , Roy M. , Dhalla N. S.
Published: December 2017

The ATPase activity of myofibrils and myosin from hindlimb muscle was investigated in animals 4 wk after the induction of diabetes by an intravenous injection of streptozotocin (65 mg/kg). Ca2+-stimulated ATPase in myofibrils was increased in diabetic muscle at various times of incubation (1-7 min) as well as at different concentrations of free Ca2+ (10(-7)-10(-5) M Ca2+). Such an increase in Ca2+-stimulated ATPase was evident as early as 1 wk after streptozotocin injection, but Mg2+-ATPase activity remained unaltered. Treatment of diabetic animals with insulin Ca2+-ATPase and actin-activated ATPase activities of pure myosin were similarly increased in diabetic muscle. Myosin ATPase was also activated by K+- or NH4+-EDTA; these responses were more in diabetic muscle. However, sodium dodecyl sulfate gel electrophoresis failed to reveal differences in the patterns of contractile proteins, and pyrophosphate gels did not show significant changes in myosin isozyme patterns between diabetics and controls. The results of this study demonstrate an activation of contractile protein ATPase of skeletal muscle in diabetes and seem to indicate that such an alteration may be responsible for enhanced contractile function of skeletal muscle in this disease.
Regulation of urea production by glucose infusion in vivo
Authors: Jahoor F. , Wolfe R. R.
Published: December 2017

We have investigated the acute in vivo regulation of urea production in normal postabsorptive volunteers by administering a primed constant infusion of 15N2-urea to measure urea production during the constant intravenous infusion of equivalent molar quantities of exogenous nitrogen, given as alanine or glutamine, either with or without a simultaneous infusion of glucose at 4 mg.kg-.min-1. These responses were compared with the response to the infusion of glucose alone. Both amino acid infusions elicited significant (P less than 0.05) and identical (26%) increases in urea production over 4 h. When the glucose infusion was added to the amino acid infusions, urea production remained constant, despite the comparable increases in plasma total nonessential amino nitrogen, as were observed with the amino acid infusions alone. Glucose infused alone elicited a significant (P less than 0.05) reduction (18%) in urea production but no corresponding change in plasma total amino nitrogen. We conclude that 1) infused glucose or its hormonal response suppresses urea production by blunting the normal hepatic ureagenic response to a fixed nitrogen load, 2) this suppressive effect is not mediated via a reduction in substrate (nitrogen) supply, and 3) the inhibition of hepatic gluconeogenesis from amino acids represents one component of this suppressive effect, and direct suppression of urea cycle activity probably represents another component.
Adrenergic blockade does not abolish elevated glucose turnover during bacterial infection
Authors: Hargrove D. M. , Bagby G. J. , Lang C. H. , Spitzer J. J.
Published: December 2017

Infusions of adrenergic antagonists were used to investigate the role of catecholamines in infection-induced elevations of glucose kinetics. Infection was produced in conscious catheterized rats by repeated subcutaneous injections of live Escherichia coli over 24 h. Glucose kinetics were measured by the constant intravenous infusion of [6-3H]- and [U-14C]glucose. Compared with noninfected rats, infected animals were hyperthermic (+1.4 degrees C) and showed increased rates of glucose appearance (45%), clearance (43%), and recycling (140%) as well as mild hyperlacticacidemia. Plasma catecholamine concentrations were increased by 50-70% in the infected rats, but there were no differences in plasma glucagon, corticosterone, and insulin levels. Adrenergic blockade was produced by primed constant infusion of both propranolol (beta-blocker) and phentolamine (alpha-blocker). A 2-h administration of adrenergic antagonists did not attenuate the elevated glucose kinetics or plasma lactate concentration in the infected rats, although it abolished the hyperthermia. In a second experiment, animals were infused with propranolol and phentolamine beginning 1 h before the first injection of E. coli and throughout the course of infection. Continuous adrenergic blockade failed to attenuate infection-induced elevations in glucose kinetics and plasma lactate. These results indicate that the adrenergic system does not mediate the elevated glucose metabolism observed in this mild model of infection.
Parathyroid hormone-independent regulation of 1,25(OH)2D in response to inhibition of bone resorption
Authors: Bonjour J. P. , Trechsel U. , Taylor C. M. , Fleisch H.
Published: December 2017

Both plasma level of 1,25-dihydroxyvitamin D [1,25(OH)2D] and intestinal Ca absorption increase after biphosphonate-induced inhibition of bone resorption. Parathyroid hormone (PTH) has been considered a key mediating element of this homeostatic response. In the present work, the role of PTH was assessed by studying the influence of 1-hydroxypentane-1,1-bisphosphonate (HPeBP) on vitamin D and Ca metabolism in both intact and thyroparathyroidectomized (TPTX) rats. In intact rats, HPeBP given at 0.1 mg P/kg body wt sc for 10 days strongly inhibited bone resorption without affecting bone formation. This effect was associated with a marked stimulation of intestinal Ca absorption and Ca balance. In this condition, HPeBP caused a marked rise in plasma 1.25(OH)2D without affecting the level of 25-hydroxyvitamin D. In TPTX rats, HPeBP given at same dose also inhibited bone resorption and enhanced plasma 1,25(OH)2D, intestinal Ca absorption and Ca balance. In summary, this study shows that bisphosphonates such as HPeBP with prevailing inhibitory activity on bone resorption induce a marked stimulation of both 1,25(OH)2D production and intestinal Ca absorption. This homeostatic response is not attenuated after PTH removal. Thus, as previously shown for the response to low Ca diet, PTH does not appear to be an essential mediating factor for stimulating 1,25(OH)2D production in response to an increase in bone mineral retention.
Epinephrine-induced renin secretion is not initiated by cardiac adrenoceptors
Authors: Johnson M. D. , Whitener C. J. , Sears T. S.
Published: December 2017

Experiments were designed to test the hypothesis that epinephrine may act directly on cardiac or pulmonary adrenoceptors to alter the release of a humoral substance that in turn influences renin secretion. Accordingly, anesthetized dogs were prepared with catheters for infusion of epinephrine at three sites: 1) into the aorta just distal to the left subclavian artery, 2) into the left ventricle, and 3) near the right atrium. Left renal renin secretion rates were determined before, during, and after 30-min infusions of epinephrine at each site in each animal; the order of the infusions was randomized. At epinephrine infusion rates of 15 and 75 ng.kg-1.min-1, epinephrine-induced changes in renin secretion rates were dose dependent but were independent of the site of infusion. These data do not support the hypotheses that either pulmonary or cardiac adrenoceptors are involved in the initiation of epinephrine-induced renin secretion. In additional experiments, an isolated canine heart was perfused with femoral arterial blood from an experimental dog, and the coronary venous effluent was returned to the experimental dog via the femoral vein. Intravenous epinephrine infusion at 50 ng.kg-1.min-1 increased plasma renin activity nearly 1.5-fold. In contrast, plasma renin activity did not increase during infusion of epinephrine at 5 ng.kg-1.min-1 directly into the coronary perfusate of the isolated heart. Coronary perfusate epinephrine concentration was 699 +/- 71 pg/ml (mean +/- SE) during intravenous infusion and was 851 +/- 121 pg/ml during direct infusion of epinephrine into the coronary perfusate. We conclude that cardiac adrenoceptors do not participate in the initiation of epinephrine-induced renin secretion.
Opposite effects of 1,25(OH)2D3 on synthesis and release of PTH compared with secretory protein I
Authors: Ridgeway R. D. , MacGregor R. R.
Published: December 2017

Aggregates of bovine parathyroid cells (organoids) were cultured with or without 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and examined for effects on the synthesis, cellular content, and release of parathyroid hormone (PTH) and secretory protein I. Organoids cultured with 120 pM 1,25(OH)2D3 at 1.35 mM calcium contained 27 +/- 2% (mean +/- SE) less and released 26 +/- 7% less PTH and contained 21 +/- 2% more and released 49 +/- 16% more secretory protein I than untreated organoids. After 15-min incubations with radioactive leucine, treated organoids incorporated 25 +/- 2% less radioactivity into precipitable cellular proteins than did untreated organoids, indicating that the overall synthetic rate of proteins was decreased by 1,25(OH)2D3 at 1.35 mM calcium. After separation of cellular proteins by gel electrophoresis, analysis of individual protein bands indicated that the mean treated-control ratio for proparathyroid hormone radioactivity was 0.88 +/- 0.06 (P less than 0.10) while that for secretory protein I was 1.56 +/- 0.17 (P less than 0.01), respectively. Experiments were performed at 1.2, 12, 120, and 1,200 pM 1,25(OH)2D3 and both 1.4 and 1.8 mM calcium. In both cases, increasing levels of 1,25(OH)2D3 progressively decreased PTH synthesis and release and progressively increased that of secretory protein I, significant effects being observed at 1.2 pM 1,25(OH)2D3. We conclude that at physiological calcium levels, 1.2-1,200 pM 1,25(OH)2D3 suppresses the synthesis and secretion of PTH, increases that of secretory protein I, and at similar concentrations decreases the overall protein synthetic rate compared with untreated controls.
Biological activity of 1,25-dihydroxyvitamin D2 and 24-epi-1,25-dihydroxyvitamin D2
Authors: DeLuca H. F. , Sicinski R. R. , Tanaka Y. , Stern P. H. , Smith C. M.
Published: December 2017

The biological activity of 1,25-dihydroxyvitamin D2 [1,25(OH)2D2] and 24-epi-1,25-dihydroxyvitamin D2 [24-epi-1,25(OH)2D2] has been determined in vitamin D-deficient rats. The biological effectiveness of 1,25(OH)2D2 is equal to that reported previously for 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] (15) in intestinal calcium transport, mineralization of bone, mobilization of bone calcium, and elevation of plasma inorganic phosphorus of rachitic rats. However, 24-epi-1,25(OH)2D2 is at best one-half as active as 1,25(OH)2D2 in stimulating intestinal calcium transport and in the mineralization of rachitic bone. The 24-epi-1,25(OH)2D2 is one-third as active as 1,25(OH)2D3 in binding to the chick intestinal receptor for 1,25(OH)2D3. Thus receptor discrimination may account for the twofold difference in intestinal calcium transport activity. 24-Epi-1,25(OH)2D2 appeared inactive in in vivo mobilization of bone calcium or bone phosphorus. On the other hand, in fetal rat bone in culture, the epi compound was only five times less active than 1,25(OH)2D2 in inducing resorption. Short-term experiments on bone mineral mobilization in vivo show that the 24-epi-1,25(OH)2D2 does induce bone calcium mobilization but that its activity in this respect is short lived. It is suggested that 24-epi-1,25(OH)2D2 and, as a result, it shows preferential activity on intestine whose response to a single dose of 1,25(OH)2D2 remains for several days, whereas the short-lived bone system does not remain stimulated during the 24-h period between doses.
Decreased autophosphorylation of EGF receptor in insulin-deficient diabetic rats
Authors: Okamoto M. , Kahn C. R. , Maron R. , White M. F.
Published: December 2017

We have previously reported that despite an increase in receptor concentration, there is a decrease in autophosphorylation and tyrosine kinase activity of the insulin receptor in insulin-deficient diabetic rats. To determine if other tyrosine kinases might be altered, we have studied the epidermal growth factor (EGF) receptor kinase in wheat germ agglutinin-purified, Triton X-100-solubilized liver membranes from streptozotocin (STZ)-induced diabetic rats and the insulin-deficient BB rat. We find that autophosphorylation of EGF receptor is decreased in proportion to the severity of the diabetic state in STZ rats with a maximal decrease of 67% (P less than 0.01). A similar decrease in autophosphorylation was observed in diabetic BB rats that was partially normalized by insulin treatment. Separation of tryptic phosphopeptides by reverse-phase high-performance liquid chromatography revealed a decrease in labeling at all sites of autophosphorylation. A parallel decrease in EGF receptor phosphorylation was also found by immunoblotting with an anti-phosphotyrosine antibody. EGF receptor concentration, determined by Scatchard analysis of 125I-labeled EGF binding, was decreased by 39% in the STZ rat (P less than 0.05) and 27% in the diabetic BB rat (not significant). Thus autophosphorylation of EGF receptor, like that of the insulin receptor, is decreased in insulin-deficient rat liver. In the case of EGF receptor, this is due in part to a decrease in receptor number and in part to a decrease in the specific activity of the kinase.(ABSTRACT TRUNCATED AT 250 WORDS)
Peptide YY antagonizes beta-adrenergic-stimulated release of insulin in dogs
Authors: Greeley G. H. , Lluis F. , Gomez G. , Ishizuka J. , Holland B. , Thompson J. C.
Published: December 2017

Peptide YY (PYY) and neuropeptide Y (NPY) are peptides of 36 amino acids that share structural homologies with pancreatic polypeptide (PP). PP is predominantly found in the endocrine pancreas. PPY is primarily found in mucosal endocrine cells of the distal ileum, colon, and rectum, whereas NPY is found in both the peripheral and central nervous systems. Previous studies indicate that these peptides can interact with the autonomic nervous system. The objective of the present experiments was to study the effect of PYY on neurally stimulated insulin release [i.e., in response to 2-deoxy-D-glucose (2-DG), a nonmetabolizable glucose analogue] in conscious dogs. Intravenous administration of PYY (100, 200, and 400 pmol.kg-1.h-1) reduced 2-DG-stimulated insulin release in a dose-dependent manner (P less than 0.05) without affecting plasma glucose levels. Administration of NPY (800 pmol.kg-1.h-1), but not PP (400 pmol.kg-1.h-1), reduced 2-DG-stimulated release of insulin (P less than 0.05). The inhibitory action of PYY on 2-DG-stimulated insulin release persisted in the presence of atropine or phentolamine treatment; however, hexamethonium alone or phentolamine plus propranolol treatment blocked the inhibitory action of PYY. Release of insulin stimulated by the beta-agonist isoproterenol was also inhibited by PYY (P less than 0.05). These results indicate that PYY can inhibit autonomic neurotransmission by a mechanism that may involve ganglionic or postganglionic inhibition of beta-adrenergic stimulation. Our findings suggest a role for PYY and NPY in the autonomic regulation of insulin release.
Effects of lack of exercise on insulin secretion and action in trained subjects
Authors: King D. S. , Dalsky G. P. , Clutter W. E. , Young D. A. , Staten M. A. , Cryer P. E. , Holloszy J. O.
Published: December 2017

We employed the hyperglycemic clamp technique to investigate the effects of short-term inactivity on insulin secretion in nine (8 men, 1 woman) well-trained subjects. A 3-h hyperglycemic clamp (plasma glucose approximately 180 mg/100 ml) was performed approximately 16 h after a usual training bout and again 14 days after stopping exercise training. There was no significant change in body composition during this short period of inactivity. The mean plasma insulin response to an identical glycemic stimulus was 67% higher after 14 days without exercise (45 +/- 7 after vs. 27 +/- 4 microU/ml before stopping exercise training). Marked increases in the early (0-10 min, 150 +/- 28 vs. 101 +/- 15 microU.ml-1.min) and late (10-180 min, 6,051 +/- 1,257 vs. 3,521 +/- 749 microU.ml-1.min) incremental insulin areas were observed as a result of the physical inactivity. Incremental areas for C-peptide were also elevated significantly in the inactive state for early (12 +/- 2.0 vs. 7 +/- 1 ng.ml-1.min) and late (567 +/- 90 vs. 467 +/- 85 ng.ml-1.min) phases. Urinary excretion of C-peptide increased from 12.1 +/- 1.5 ng/240 min in the exercising state to 21.8 +/- 3.6 ng/240 min in the inactive state. Rates of whole body glucose disposal were not different between exercising and inactive states, indicating a large increase in resistance to the action of insulin. These findings indicate that the decreased insulin secretory response to a glucose stimulus in people who exercise regularly is a relatively short-term effect of exercise.
Uptake of exogenous substrates during hypoxia in fetal lambs
Authors: Milley J. R.
Published: December 2017

Fetal uptakes of oxygen, glucose, lactate, and alpha-amino-nitrogen containing substrates were measured in eight near-term fetal lambs by simultaneously drawing blood samples from the umbilical vein and descending aorta, then measuring umbilical blood flow using the microsphere method. These procedures were repeated after 3 h of hypoxia induced by lowering maternal inspired oxygen concentration. On the next day the experiment was repeated, except the ewes were first made hypoxic then allowed to breathe room air. These conditions decreased the delivery of oxygen, but not the delivery of other metabolic substrates to the fetus. During hypoxia, fetal oxygen uptake was 82% of normal (mean of both days); fetal glucose and amino-nitrogen uptakes were 74 and 23% of normal, respectively, and fetal lactate uptake became insignificant. These data indicate that endogenous rather than exogenous substrates are used to support fetal oxidative metabolism during hypoxia. Also, because exogenous uptake of amino-nitrogen is less than normal nitrogen accretion rates, fetal growth must be reduced as a consequence of 3-4 h of hypoxia.
Modulatory effect of glucose on VIP-induced gastric somatostatin release
Authors: Schmid R. , Schusdziarra V. , Classen M.
Published: December 2017

The present study was designed to examine the effect of increasing perfusate glucose concentrations on vasoactive intestinal peptide (VIP)-induced somatostatin (SLI) release from the isolated rat stomach. The stomach of overnight-fasted rats was perfused with Krebs-Ringer buffer containing 100, 150, or 200 mg/dl glucose, respectively. VIP was administered at 10(-12), 10(-11), 10(-9), and 10(-8) M. At a normal glucose concentration of 100 mg/dl, VIP at doses of 10(-12), 10(-11), and 10(-9) M elicited a small inhibitory effect on SLI release by 200-300 pg/min (P less than 0.01). As reported previously at 10(-8) M, VIP stimulated gastric SLI secretion by 500 pg/min (P less than 0.01). Increasing perfusate glucose to 150 mg/dl resulted in a stimulation of SLI release by all four concentrations of VIP with a maximal effect at 10(-9) M. During 200 mg/dl glucose, VIP had no effect in concentrations below 10(-9) M, and only the two highest doses (10(-9) and 10(-8) M) stimulated SLI release significantly. In the absence of VIP, glucose had no effect on gastric SLI release. In conclusion, the present data demonstrate for the first time that at normal glucose levels VIP has not only stimulatory but also inhibitory effects on gastric SLI, and second, a modest elevation of glucose has a modulatory effect on gastric D-cell function.
Impaired transport of thyroid hormones into livers of obese (ob/ob) mice
Authors: Hillgartner F. B. , Romsos D. R.
Published: December 2017

Obese (ob/ob) mice exhibit impaired hepatic thyroid hormone action that is mediated, at least in part, by a reduced nuclear 3,5,3'-triiodothyronine (T3) receptor occupancy. The possibility that lowered occupancy in obese mice may be caused by decreased transport of T3 across the hepatic plasma membrane was examined by measuring the unidirectional influx of [125I]T3 into livers of 8- to 10-wk-old obese and lean mice using a tissue-sampling portal vein-injection technique. Influx of [125I]thyroxine (T4), a substrate for T4 5'-deiodinase, was also measured. Unidirectional clearance of T3 and T4 was 64 and 80% lower, respectively, in obese mice than in lean mice. Hepatic T3 and T4 uptake was nonsaturable in both lean and obese mice, suggesting that transport occurs by lipid-mediated free diffusion. Clearance of another lipid-soluble hormone, hydrocortisone, was also lower in obese mice than in lean mice. Decreased membrane permeability to the above hormones in obese mice may result from reported changes in membrane lipid composition. In conclusion, decreased hepatic thyroid hormone uptake may contribute to impaired thyroid hormone action and T3 production in livers of obese mice.
Pancreatic islet discrimination of hexose anomers. II. Transient computer simulation
Authors: Achs M. J. , Garfinkel D.
Published: December 2017

We have previously modeled pancreatic islet glycolysis under idealized steady-state conditions where the input is a pure hexose anomer and there is no mutarotation and reproduced the known preference for the alpha-anomers of glucose and mannose as substrates. This model is here extended to simulate real experiments, where the hexoses mutarotate and measurements may be taken over time. The behavior of our model system agrees with available experimental data. The hexose diphosphate activators of phosphofructokinase, whose effect was seen as not important in the preceding steady-state analysis, are found here to have a modest (approximately 10–15%) effect on its flux. The previous conclusion that the anomeric preference of the glycolytic pathway follows from that of glucokinase continues to hold in the real experimental situation.
Effect of postweaning thyroid status on endocrine and adipose development in rats
Authors: Robert M. Q. , Herrera E.
Published: December 2017

We previously reported that 6 wk after thyroidectomy, rat fat cell diameter and number were 86 and 32% of values in age-matched controls. This study aims to 1) determine the time course of these changes and of parallel changes in the levels of several hormones known to participate in adipose differentiation and 2) try to normalize the altered cellularity with thyroxine (T4) treatment. Rats were thyroidectomized (T) or sham-operated and studied 2, 5, or 6 wk later. At 5 wk some T rats received L-T4. Epididymal fat cells were sized and used to estimate fat cell number. Triiodothyronine (T3), T4, thyroid-stimulating hormone, somatotropin (GH), and insulin plasma levels were assayed. In T rats fat cell hypertrophia was blunted uniformly with time, whereas fat cell hyperplasia was slow for 2 wk and zero thereafter. Insulinemia was less drastically reduced than T4, T3, and GH levels. The fat cell number-to-body weight ratio always stayed close to the ratio present at thyroidectomy. In T rats receiving a T4 substitution dose for 1 wk, fat cell number and body weight increased, but not when the T4 dose was lower even though the plasma thyroid hormone levels were increased. These thyroid hormone effects on adipose tissue are probably secondary to other endocrine changes that occur in hypothyroidism.
Glucose transport: locus of muscle insulin resistance in obese Zucker rats
Authors: Sherman W. M. , Katz A. L. , Cutler C. L. , Withers R. T. , Ivy J. L.
Published: December 2017

The purposes of this study were to determine whether the muscle insulin resistance of the obese rat is due to a defect in the glucose transport process and whether the insulin resistance is fiber-type specific. The hindlimbs of fasted, 14-wk-old obese (fa/fa) and lean (fa/?) Zucker rats were perfused with perfusate containing 8 mM glucose and no insulin or 8 mM glucose and either a physiological (0.15 mU/ml), a submaximal (1.50 mU/ml), or a maximal (15.0 mU/ml) insulin concentration. Glucose uptake was determined after which the initial rate of glucose transport was determined using 3-O-methyl-D-glucose (3-OMG). Glucose uptake of the obese rats was depressed by 40, 33, 42, and 47% in the absence of insulin and in the presence of the physiological, submaximal, and maximal insulin concentrations, respectively, when compared with lean littermates. Glucose transport in the absence and in the presence of the three insulin concentrations was significantly lower in the soleus (slow-twitch, oxidative fibers), red quadriceps (fast-twitch, oxidative, glycolytic fibers), and gastrocnemius (mixed fibers) of the obese rats when compared with lean rats. Glucose transport in the white quadriceps (fast-twitch, glycolytic fibers) was significantly lower in the obese rats in the absence of insulin and in the presence of the submaximal and maximal insulin concentrations. The glycogen concentration and the activity of hexokinase were the same and the glycogen synthase activity was higher in the muscles for the obese rats when compared to lean rats.(ABSTRACT TRUNCATED AT 250 WORDS)
Role of Ca2+ and Na+ on luteinizing hormone release from the calf pituitary
Authors: Kile J. P. , Amoss M. S.
Published: December 2017

It has been proposed that gonadotropin-releasing hormone (GnRH) stimulates Ca2+ entry by activation of voltage-independent, receptor-mediated Ca2+ channels in the rat gonadotroph. Little work has been done on the role of calcium in GnRH-induced luteinizing hormone (LH) release in species other than the rat. Therefore, this study was done to compare the effects of agents that alter Ca2+ or Na+ entry on LH release from calf anterior pituitary primary cells in culture. GnRH (100 ng/ml), Ca2+ ionophore A23187 (2.5 microM), and the depolarizing agent ouabain (0.1-10 microM) all produced significant increases (P less than 0.05) in LH release; these effects were significantly reduced when the cells were preincubated with the organic Ca2+ channel blockers nifedipine (1-10 microM) and verapamil (1-10 microM) and with Co2+ (0.01-1 mM). The effect of ouabain was inhibited by tetrodotoxin (TTX; 1-10 nM) as well as by nifedipine at 0.1-10 microM. In contrast to its effect on rat pituitary LH release, TTX significantly inhibited GnRH-stimulated LH release at 1-100 nM. These results suggest that GnRH-induced LH release may employ Ca2+ as a second messenger in bovine gonadotrophs and support recent speculation that GnRH-induced Ca2+ mobilization may in part be voltage dependent.
Regulation of valine and alpha-ketoisocaproate metabolism in rat kidney mitochondria
Authors: Miller R. H. , Harper A. E.
Published: December 2017

Activities of branched-chain amino acid (BCAA) aminotransferase (BCAT) and alpha-keto acid dehydrogenase (BCKD) were assayed in mitochondria isolated from kidneys of rats. Rates of transamination of valine and oxidation of keto acids alpha-ketoisocaproate (KIC) or alpha-ketoisovalerate (KIV) were estimated using radioactive tracers of the appropriate substrate from amounts of 14C-labeled products formed (14CO2 or [1-14C]-keto acid). Because of the high mitochondrial BCAT activity, an amino acceptor for BCAT, alpha-ketoglutarate (alpha-KG) or KIC, was added to the assay medium when valine was the substrate. Rates of valine transamination and subsequent oxidation of the KIV formed were determined with 0.5 mM alpha-KG as the amino acceptor; these rates were 5- to 50-fold those without added alpha-KG. Rates of CO2 evolution from valine also increased when KIC (0.01-0.10 mM) was present; however, with KIC concentrations above 0.2 mM, rates of CO2 evolution from valine declined although rates of transamination continued to rise. When 0.05 mM KIC was added to the assay medium, oxidation of KIC was suppressed by inclusion of valine or glutamate in the medium. When valine was present KIC was not oxidized preferentially, presumably because it was also serving as an amino acceptor for BCAT. These results indicate that as the supply of amino acceptor, alpha-KG or KIC, is increased in mitochondria not only is the rate of valine transamination stimulated but also the rate of oxidation of the KIV formed from valine.(ABSTRACT TRUNCATED AT 250 WORDS)
Pituitary-adrenal response to bacterial endotoxin in developing rats
Authors: Witek-Janusek L.
Published: December 2017

The neonatal rat is very sensitive to the lethal effects of bacterial endotoxin. Because of the adaptive importance of pituitary-adrenal secretions to stress, this study examined the ontogeny of the plasma corticosterone and adrenocorticotropic hormone (ACTH) responses to endotoxin. The lethal sensitivity of young rats to endotoxin ranged from 0.5 to 30 mg/kg (ip) in the 1- to 21-day-old rat. After endotoxin treatment, the 1- and 2-day-old rat showed marked elevations of corticosterone similar in magnitude to that seen in 21-day-old and adult rats; however, significantly depressed corticosterone increments were observed in the 5-, 10-, and 14-day-old rats. This age-related pattern of adrenocortical secretion was correlated with the developing rat's corticosterone response to exogenous ACTH. In contrast, endotoxin administered to 5-, 10-, and 14-day-old rats resulted in increments of plasma ACTH similar to those observed in the 21-day-old and adult rats. Although plasma ACTH levels increased by 84-127% in the 1- and 2-day-old rats, these increases were significantly less than those of rats at all other ages tested. Thus the newborn rat mounts an effective corticosterone response to endotoxin, loses this ability between ages 5-14 days, and regains this response at 21 days of age. Because the hyporesponsive ages exhibit a marked increase in ACTH secretion, the loss of the adrenocortical response to endotoxin appears to be a result of a depressed responsiveness of the adrenal cortex to ACTH.
Effect of prolactin on enzymes of lipid biosynthesis in mammary gland explants
Authors: Waters S. B. , Rillema J. A.
Published: December 2017

Prolactin (PRL) stimulates an increased rate of incorporation of [14C]acetate and [3H]glucose into lipids in cultured mammary gland explants from 10- to 14-day-pregnant mice. This response is biphasic with an early increase occurring from 6 through 12 h, and an additional increase from 16 through 24 h. Enzymes likely to be rate limiting to this process include acetyl CoA carboxylase, fatty acid synthetase, acetyl CoA synthetase, and/or pyruvate dehydrogenase. Of these enzymes only pyruvate dehydrogenase activity was elevated at 6 h, suggesting that this enzymatic activity is important in stimulating early increases in lipogenesis after PRL treatment. In addition, the PRL stimulation of pyruvate dehydrogenase may also indirectly stimulate acetyl CoA carboxylase through the generation of citrate; this may explain the early (6-12 h) effect of PRL on [14C]acetate incorporation. After 16 h of PRL treatment, the activities of all the lipogenic enzymes were enhanced. The second phase of PRLs stimulation of lipogenesis thus likely involves the enhanced activities of more than one of the lipogenic enzymes.
Adrenalectomy fails to stimulate brown adipose tissue metabolism in ob/ob mice fed glucose
Authors: Kim H. K. , Romsos D. R.
Published: December 2017

Adrenalectomy arrests the development of obesity in ob/ob mice fed nonpurified high-starch diets partly by stimulating the low thermogenic activity of brown adipose tissue (BAT). However, adrenalectomy fails to suppress the development of obesity in ob/ob mice fed a purified high-glucose diet. Effects of adrenalectomy on BAT metabolism in ob/ob mice fed purified high-starch or high-glucose diets were therefore examined. Adrenalectomy markedly decreased the efficiency of energy retention and increased BAT metabolism (as assessed by GDP binding to BAT mitochondria, GDP-inhibitable acetate- or chloride-induced mitochondrial swelling, and by rates of norepinephrine turnover in BAT) in ob/ob mice fed a high-starch purified diet but had only minimal effects on energy efficiency or BAT metabolism in ob/ob mice fed a high-glucose purified diet. Plasma insulin concentrations decreased and thyroxine concentrations increased in adrenalectomized ob/ob mice fed the high-starch diet; changes in these hormones were less pronounced in adrenalectomized ob/ob mice fed the high-glucose diet. Consumption of glucose mimics effects of adrenal secretions on BAT metabolism in ob/ob mice.
Nonadrenergic sympathetic neural influences on basal pancreatic hormone secretion
Authors: Dunning B. E. , Ahren B. , Veith R. C. , Taborsky G. J.
Published: December 2017

Evidence for peptidergic innervation of the islets of Langerhans is increasing, yet the role of neuropeptides in mediating neurally induced changes of islet function is not clear. To determine if nonadrenergic transmitters make an important contribution to sympathetic neural effects on basal pancreatic hormone secretion, we examined the effect of local sympathetic nerve stimulation (SNS) on the output of immunoreactive insulin (IRI), immunoreactive glucagon (IRG), and somatostatin (SLI) from the duodenal lobe of the pancreas in situ in halothane-anesthetized dogs, under conditions where the actions of the classical transmitter norepinephrine (NE) should be blocked by propranolol (PROP) and yohimbine (YO). In the absence of adrenergic antagonists, SNS rapidly reduced the output of IRI (delta = -1.34 +/- 0.91 mU/min) and SLI (delta = -600 +/- 350 fmol/min) and stimulated that of IRG (delta = +1.39 +/- 0.57 ng/min). In the presence of PROP and YO, SNS induced similar changes of hormone secretion: delta IRI, -1.30 +/- 0.53 mU/min; delta SLI, -480 +/- 180 fmol/min; delta IRG = +1.89 +/- 0.63 ng/min. Because PROP and YO abolished the pancreatic effects of high dose infusions of NE (1 microgram.kg-1.min-1 iv), we suggest that the antagonists produced sufficient, combined adrenergic blockade at the level of the islet, and we conclude that a nonadrenergic neurotransmitter or modulator plays a major role in mediating sympathetic neural effects on basal islet hormone secretion.