bluuebunny/crossref_metadata_2025 huggingface dataset:
Central catecholaminergic system stimulates secretion of CRH at different sites
Authors: Szafarczyk A. , Guillaume V. , Conte-Devolx B. , Alonso G. , Malaval F. , Pares-Herbute N. , Oliver C. , Assenmacher I.
Published: December 2017

To explore a possible differential role of distinct catecholamine (CA) innervation sites in corticotropin-releasing hormone (CRH) secretion, especially under stress conditions, we compared the effects in adult female rats of selective CA denervation of either the whole hypothalamus, by a discrete pharmacological lesion of the ventral noradrenergic ascending bundle [VNAB; 3 micrograms of 6-hydroxydopamine (6-OHDA) in 0.2 microliter of vehicle, bilaterally] or of the paraventricular nuclei (PVN) alone (1 microgram of 6-OHDA in 0.2 microliter of vehicle, bilaterally). Although both procedures induced a similar dramatic fall in norepinephrine and epinephrine concentrations (-55 to -65%) measured by high-performance liquid chromatography in PVN punches, the VNAB lesion, unlike PVN denervation, depleted the median eminence (ME) of both amines (-80%). Concomitantly, the VNAB lesion led to a 97% reduction of the immunoreactive (ir) CRH-41 concentration in the hypophysial portal vessels, associated with a 64% fall in plasma adrenocorticotropic hormone (ACTH), and, in another group, with an 80% inhibition of ether stress-induced ACTH surge. The deletion of CA innervation of the PVN alone reduced irCRH-41 levels in the portal vessels by only 57% and plasma ACTH by 35%. This lesion did not significantly impair stress-induced ACTH release. These results suggest that the CA innervation of the hypothalamus exerts a stimulatory control on CRH-41-secreting neurons not only directly at the perikaryal level but also at other hypothalamic sites of VNAB innervation including peripheral contacts between the terminals of CA and CRH nerves in the external ME.
Impact of sex steroids and their suppression on skeletal growth and maturation
Authors: Boepple P. A. , Mansfield M. J. , Link K. , Crawford J. D. , Crigler J. F. , Kushner D. C. , Blizzard R. M. , Crowley W. F.
Published: December 2017

Forty girls with central precocious puberty (CPP) were studied before and during 1-3 yr of luteinizing hormone-releasing factor (LHRH) agonist (LHRHa) administration to examine the impact of gonadal steroid secretion and its suppression on skeletal growth and maturation. Pubertal growth velocity (GV) was 10.1 +/- 0.7 (SE) cm/yr and, when normalized for chronological age (CA) and bone age (BA), demonstrated that the effects of sex steroids were most profound in patients with the youngest CA and BA. GV decreased significantly to 5.8 +/- 0.3 (n = 40), 4.6 +/- 0.3 (n = 30), and 3.2 +/- 0.6 cm/yr (n = 12) during 3 yr of gonadal suppression and correlated negatively with starting BA. Skeletal maturation was markedly accelerated by premature sex steroid secretion (BA/CA = 1.8 +/- 0.1), was slowed significantly with gonadal suppression (mean delta BA/delta CA less than 1), and also was negatively correlated with the starting BA. Cumulative increases in predicted adult height were observed regardless of starting BA and averaged +2.0 +/- 0.4, +5.2 +/- 0.5, and +6.7 +/- 1.2 cm after 1, 2, and 3 yr of gonadal suppression. The comparable changes in height predictions across all BAs despite highly variable GVs underscore the need for use of developmental (i.e., BA-based) rather than CA-based standards in the analysis of growth during gonadal steroid exposure and suppression in childhood.
G protein in stimulation of PI hydrolysis by CCK in isolated rat pancreatic acinar cells
Authors: Matozaki T. , Sakamoto C. , Nagao M. , Nishizaki H. , Baba S.
Published: December 2017

To clarify the possible role of a guanine nucleotide-binding protein (G protein) in the signal transducing system activated by cholecystokinin (CCK), actions of CCK on rat pancreatic acini were compared with those of fluoride, a well-known activator of stimulatory (Gs) or inhibitory (Gi) G protein. When acini were incubated with increasing concentrations of either CCK-octapeptide (CCK8) or NaF, a maximal stimulation of amylase release from acini occurred at 100 pM CCK8 or 10 mM NaF, respectively; this secretory rate decreased as CCK8 or NaF concentration was increased. NaF caused an increased in cytoplasmic Ca2+ concentration from the internal Ca2+ store and stimulated accumulation of inositol phosphates in acini, as observed with CCK. However, NaF-stimulated Ca2+ mobilization had a lag period before detectable stimulation and was potentiated by AlCl3. These stimulatory effects of NaF appeared to be independent of cellular adenosine 3',5'-cyclic monophosphate (cAMP). Pretreatment with cholera toxin or pertussis toxin did not affect CCK8- or NaF-induced inositol phosphate accumulation or Ca2+ mobilization. 5'-Guanylimidodiphosphate activated the generation of inositol phosphates in the [3H]inositol-labeled pancreatic acinar cell membrane preparation, with half-maximal and maximal stimulation at 1 and 10 microM, respectively. Furthermore, the effects of submaximal CCK concentrations on inositol phosphate accumulation in membranes were markedly potentiated in the presence of 100 microM GTP, which alone was ineffective. Combined findings of the present study strongly suggest that pancreatic CCK receptors are probably coupled to the activation of polyphosphoinositide (PI) breakdown by a G protein, which appears to be fluoride sensitive but is other than Gs- or Gi-like protein.
Effects of epinephrine infusion on determinants of intravenous glucose tolerance in dogs
Authors: Martin I. K. , Weber K. M. , Boston R. C. , Alford F. P. , Best J. D.
Published: December 2017

Effects of four- to fivefold elevations of epinephrine (EPI) on glucose (Glc) metabolism were assessed in eight dogs before and after an intravenous Glc tolerance test, performed 30 min (short EPI) and 72 h (long EPI) after start of EPI infusion. Short EPI increased plasma nonesterified fatty acids (NEFA; 0.46 +/- 0.08 to 0.78 +/- 0.12 mmol/l, P less than 0.05), but Glc and insulin were unchanged. After long EPI, NEFA returned to control but Glc increased from 5.1 +/- 0.1 to 5.7 +/- 0.2 mmol/l (P less than 0.05). EPI reduced overall Glc tolerance (KG) from 3.5 +/- 0.7 to 2.5 +/- 0.2 (short EPI, P less than 0.05) and 2.3 +/- 0.3%/min (long EPI, P less than 0.02). Minimal model analysis showed that short EPI decreased insulin sensitivity (SI) from 7.9 +/- 1.1 to 4.2 +/- 1.2 min-1 per mU/l X 10(-4) (P less than 0.005) and increased pancreatic responsiveness (phi 1 from 3.7 +/- 0.3 to 7.4 +/- 2.9 mU/l.min-1 per mg/dl, P less than 0.025; phi 2 from 2.6 +/- 0.7 to 4.9 +/- 1.2 mU/l.min-2 per mg/dl). After long EPI SI, phi 1, and phi 2 returned to control. In contrast, Glc-mediated Glc disposal (SG) was decreased from 3.5 +/- 0.5 X 10(-2) to 2.8 +/- 0.6 X 10(-2) (short EPI) and 1.3 +/- 0.6 X 10(-2) min-1 (long EPI, P less than 0.02). We conclude that prolonged infusion of EPI leads to adaptation to its acute effects on NEFA, SI, phi 1, and phi 2.(ABSTRACT TRUNCATED AT 250 WORDS)
Peripheral metabolism of PTH: fate of biologically active amino terminus in vivo
Authors: Bringhurst F. R. , Stern A. M. , Yotts M. , Mizrahi N. , Segre G. V. , Potts J. T.
Published: December 2017

Clearance of intact parathyroid hormone (PTH) from blood is associated with rapid uptake by liver and kidney, limited proteolysis by tissue endopeptidases and, within minutes, appearance of circulating carboxyl-(COOH)-terminal PTH fragments. The fate of the corresponding amino(NH2)-terminal portion of the hormone during this peripheral metabolism is still unknown, however. To determine this, we have employed [35S]bovine PTH (bPTH) labeled to high specific activity at NH2-terminal methionines, which permits direct monitoring of the fate of the PTH NH2-terminus during metabolism in vivo. The [35S]PTH was administered by bolus or continuous intravenous infusion to anesthetized normal rats, to rats subjected to acute ablation of the liver, the kidneys, or both, and to rats receiving co-infusions of excess synthetic bPTH(1-34) NH2-terminal fragments. Analysis by high-resolution chromatographic techniques sensitive to 10(-13) M [35S]PTH peptides in plasma yields no evidence that peripheral metabolism of PTH generates circulating NH2-terminal fragments, even when special measures are taken to block clearance of such putative fragments from blood. We find that the NH2-terminus of PTH is rapidly degraded in situ by the liver but that both liver and especially kidney nevertheless contain low levels of NH2-terminal PTH fragments that, although not released into the blood, are large enough to be potentially active. Thus, the peripheral metabolism of PTH in normal animals does not normally lead to the formation of circulating amino terminal fragments of the hormone that might act independently of intact PTH on peripheral target tissues.
Adipose tissue and skeletal muscle blood flow during mental stress
Authors: Linde B. , Hjemdahl P. , Freyschuss U. , Juhlin-Dannfelt A.
Published: December 2017

Mental stress [a modified Stroop color word conflict test (CWT)] increased adipose tissue blood flow (ATBF; 133Xe clearance) by 70% and reduced adipose tissue vascular resistance (ATR) by 25% in healthy male volunteers. The vasculatures of adipose tissue (abdomen as well as thigh), skeletal muscle of the calf (133Xe clearance), and the entire calf (venous occlusion plethysmography) responded similarly. Arterial epinephrine (Epi) and glycerol levels were approximately doubled by stress. beta-Blockade by metoprolol (beta 1-selective) or propranolol (nonselective) attenuated CWT-induced tachycardia similarly. Metoprolol attenuated stress-induced vasodilation in the calf and tended to do so in adipose tissue. Propranolol abolished vasodilation in the calf and resulted in vasoconstriction during CWT in adipose tissue. Decreases in ATR, but not in skeletal muscle or calf vascular resistances, were correlated to increases in arterial plasma glycerol (r = -0.42, P less than 0.05), whereas decreases in skeletal muscle and calf vascular resistances, but not in ATR, were correlated to increases in arterial Epi levels (r = -0.69, P less than 0.01; and r = -0.43, P less than 0.05, respectively). The results suggest that mental stress increases nutritive blood flow in adipose tissue and skeletal muscle considerably, both through the elevation of perfusion pressure and via vasodilatation. Withdrawal of vasoconstrictor nerve activity, vascular beta 2-adrenoceptor stimulation by circulating Epi, and metabolic mechanisms (in adipose tissue) may contribute to the vasodilatation.
Fasting and postabsorptive hepatic glucose and insulin metabolism in hyperthyroidism
Authors: Raboudi N. , Arem R. , Jones R. H. , Chap Z. , Pena J. , Chou J. , Field J. B.
Published: December 2017

The effect of thyroid hormone excess on hepatic glucose balances and fractional hepatic extraction of insulin and glucagon was examined in six conscious dogs with catheters in the portal vein, hepatic vein, and femoral artery and Doppler flow probes on the portal vein and hepatic artery. An oral glucose tolerance test was performed before and after the animals were made hyperthyroid by intramuscular thyroxine administration (100 micrograms.kg-1.day-1) for 10 days. In the basal state and after oral glucose, insulin and glucagon levels in the three vessels and the basal fractional hepatic extraction of insulin and glucagon were not significantly modified by thyroid hormone. These results suggest that in short-term thyrotoxicosis insulin secretion is not impaired, and the rise in fasting plasma glucose and increased hepatic glucose production could reflect hepatic insulin resistance, increased availability of precursors for gluconeogenesis, or increased glycogenolysis. Hyperthyroidism significantly increased basal flows in the portal vein (14.7 +/- 0.6 vs. 12.9 +/- 0.5 ml.kg-1.min-1), the hepatic artery (4.8 +/- 0.3 vs. 3.9 +/- 0.2 ml.kg-1.min-1) and vein (19.6 +/- 0.7 vs. 16.9 +/- 0.4 ml.kg-1.min-1), the fasting plasma glucose concentration (104 +/- 3 vs. 92 +/- 2 mg/dl), and basal hepatic glucose output (2.1 +/- 0.2 vs. 1.5 +/- 0.2 mg.kg-1.min-1). It did not alter the nonhepatic splanchnic uptake of glucose, the percent of orally administered glucose that appeared in the portal vein (47 +/- 2 vs. 45 +/- 11%), the percent of hepatic uptake of glucose (59 +/- 11 vs. 74 +/- 22%), or the shape of the glucose tolerance test.
Effect of thyroid status on catecholamine stimulation of thyroxine 5'-deiodinase in brown adipocytes
Authors: Mills I. , Raasmaja A. , Moolten N. , Lemack G. , Silva J. E. , Larsen P. R.
Published: December 2017

We examined type II 5'-iodothyronine deiodinase activation by adrenergic agonists in dispersed brown adipocytes from euthyroid and hypothyroid rats. In euthyroid cells, basal deiodinase activity was 30-100 fmol I-.h-1.10(6) cells-1 and increased four- to fivefold during exposure to norepinephrine, an effect that was enhanced by alprenolol. In cells from hypothyroid rats, norepinephrine caused a three- to fourfold greater deiodinase stimulation than occurred in euthyroid cells but alprenolol inhibited the response. In euthyroid cells, phenylephrine caused greater stimulation than did norepinephrine, but this was inhibited by alprenolol. Isoproterenol and 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP) inhibited the phenylephrine response but were modestly stimulatory alone. Although both alpha 1- and beta-adrenergic agonists increased deiodinase activity modestly in hypothyroid cells, in combination they caused a marked synergistic stimulation. This synergism was induced by 8-BrcAMP and forskolin, as well as by isoproterenol. The stimulation of deiodinase in both cell types was due to an increase in Vmax without an alteration in the Km and required mRNA synthesis. The markedly greater deiodinase response of the hypothyroid brown adipocyte to catecholamines may serve to enhance the impaired thermogenic response of this tissue to cold exposure.
Insulin degradation products from perfused rat kidney
Authors: Duckworth W. C. , Hamel F. G. , Liepnieks J. , Peavy D. , Frank B. , Rabkin R.
Published: December 2017

The kidney is a major site for insulin metabolism, but the enzymes involved and the products generated have not been established. To examine the products, we have perfused rat kidneys with insulin specifically iodinated on either the A14 or the B26 tyrosine. Labeled material from both the perfusate and kidney extract was examined by Sephadex G50 and high-performance liquid chromatography (HPLC). In perfusate from a filtering kidney, 22% of the insulin-sized material was not intact insulin on HPLC. With the nonfiltering kidney, 10.6% was not intact insulin. Labeled material from HPLC was sulfitolyzed and reinjected on HPLC. By use of 125I-iodo(A14)-insulin, almost all the degradation products contained an intact A-chain. By use of 125I-iodo(B26)-insulin, several different B-chain-cleaved products were obtained. The material extracted from the perfused kidney was different from perfusate products but similar to intracellular products from hepatocytes, suggesting that cellular metabolism by kidney and liver are similar. The major intracellular product had characteristics consistent with a cleavage between the B16 and B17 amino acids. This product and several of the perfusate products are also produced by insulin protease suggesting that this enzyme is involved in the degradation of insulin by kidney.
Microdialysis of intercellular adenosine concentration in subcutaneous tissue in humans
Authors: Lonnroth P. , Jansson P. A. , Fredholm B. B. , Smith U.
Published: December 2017

To determine the intercellular adenosine concentration the periumbilical subcutaneous interstitial fluid was investigated in five healthy subjects with two single dialysis fiber catheters perfused with isotonic saline at a rate of 2.5 mul/min. A newly developed in situ calibration technique (29) allows an estimation of the adenosine concentration in the perfusate equilibrating with the surrounding medium, i.e., the concentration in the interstitial fluid. The mean interstitial adenosine concentration was 128 +/- 26 nM (range 25-300 nM). The effect of these adenosine concentrations on adipose tissue metabolism was investigated in isolated human fat cells. The influence of 5 and 10 nM 2-chloroadenosine, which is equipotent to 150 and 300 nM adenosine, on the dose-response curve for the lipolytic effect of norepinephrine and the stimulatory effect of insulin on glucose transport was studied. 2-Chloroadenosine at 10 nM decreased basal lipolysis 53 +/- 8% and shifted the dose-response curve for norepinephrine approximately twofold to the right. At 5 nM, 2-chloroadenosine only slightly shifted the dose-response curve, whereas basal lipolysis was not significantly reduced. 2-Chloroadenosine at either concentration had no significant effect on basal or insulin-stimulated glucose transport. In conclusion, endogenous adenosine can reach concentrations in human subcutaneous tissue sufficient for an important modulating effect on lipolysis in vivo.
Palmitate oxidation by isolated working fetal and newborn pig hearts
Authors: Werner J. C. , Sicard R. E. , Schuler H. G.
Published: December 2017

Palmitate oxidation and the effect of palmitate on glucose and lactate utilization were investigated in isolated, perfused, fetal (0.9 gestation), and neonatal (2 day old) pig hearts. Hearts were perfused under working conditions, developing a mean aortic pressure of 50-55 mmHg, paced at 180 beats/min for 30 min, with Krebs-Henseleit buffer containing 3% albumin, glucose (5 mM), and insulin (100 microU/ml). Palmitate (1 mM) and lactate (5 mM), either individually or in combination, were added to the perfusion buffer. Palmitate oxidation was assessed from 14CO2 production from [U-14C]-palmitate, glucose uptake as 3H2O production from D-[2-3H]-glucose, and lactate metabolism from changes in buffer lactate content. After perfusion, ATP, creatine phosphate, triglycerides, and glycogen were measured. Substantial palmitate oxidation was observed at both ages but was greater in neonatal hearts. Nevertheless, palmitate inhibited lactate utilization and glucose uptake similarly in fetal and neonatal hearts. Lactate also reduced palmitate uptake and oxidation by 40-60% in both fetal and neonatal hearts. During perfusions with palmitate, tissue concentrations of triglycerides increased approximately threefold in fetal hearts and were unaffected by lactate. Thus both palmitate and lactate can act as major energy substrates for the immature heart. Both substrates significantly (P less than 0.01) suppress glucose utilization, and each has suppressive effects on the other's metabolism.
Potassium-induced aldosterone biosynthesis in cultured rat zona glomerulosa cells
Authors: Muller J. , Lauber M. , Schmid C.
Published: December 2017

Rat adrenal zona glomerulosa cells lost their ability to produce aldosterone from either endogenous precursors or added deoxycorticosterone within 2 days of primary monolayer culture in a medium with a potassium concentration of 6.3 mmol/l. The lost corticosterone methyl oxidase I and II activities were totally regenerated when the ambient potassium concentrations was raised to 31 mmol/l. The conversions of deoxycorticosterone to 18-hydroxycorticosterone and aldosterone were completely restored by culture in a high-potassium medium also in zona glomerulosa cells of rats in which aldosterone biosynthesis had been suppressed by potassium restriction and sodium loading. However, these conversions were not induced in zona fasciculata-reticularis cells. The induction of aldosterone biosynthesis was associated with the appearance of a mitochondrial 49,000 protein cross-reacting with an antibody raised against bovine adrenal cytochrome P-450(11) beta. Thus primary cultures of zona glomerulosa cells are promising models for studying in vitro the molecular mechanisms of long-term adaptation of aldosterone biosynthesis to sodium and potassium intake.
Rapid measurement of whole body and forearm protein turnover using a [2H5]phenylalanine model
Authors: Thompson G. N. , Pacy P. J. , Merritt H. , Ford G. C. , Read M. A. , Cheng K. N. , Halliday D.
Published: December 2017

Whole body protein turnover was measured in six normal adults using a model based on a primed constant infusion of [2H5]phenylalanine and, independently, by an established method of a primed constant infusion of [1-13C]leucine. Isotopic plateau in plasma was achieved within 2 h for [2H5]phenylalanine and, in four of the subjects who received a priming dose of [2H4]tyrosine, for [2H4]tyrosine. In all subjects whole body protein turnover measured with the phenylalanine model (mean protein synthesis, 2.65 +/- (SD) 0.16 g.kg-1.24 h-1; catabolism, 3.58 +/- 0.26 g.kg-1.24 h-1) was similar to that measured using the leucine model (synthesis, 3.09 +/- 0.27 g.kg-1.24 h-1; catabolism, 3.70 +/- 0.35 g.kg-1.24 h-1). Mean forearm fractional muscle protein synthesis calculated by the phenylalanine model was 0.06 +/- 0.03%/h, which compares closely with literature values derived by other methods. The phenylalanine model allows the rapid assessment of whole body and muscle protein turnover from plasma samples alone, obviating the need for measurement of expired air CO2 production or enrichment.
Increase in ovarian leukotrienes during hormonally induced ovulation in the rat
Authors: Espey L. L. , Tanaka N. , Okamura H.
Published: December 2017

The ovulatory process was initiated in 25-day-old Wistar rats by injecting human chorionic gonadotropin (hCG; 10 IU sc) 2 days after the animals had been primed with pregnant mares serum gonadotropin (PMSG; 10 IU sc). By 4 h into the ovulatory process, leukotriene (LT) B4 increased 2-fold (P less than 0.001) and LTs C4/D4/E4 increased 1.3-fold (P less than 0.002). By the time of ovulation (10-12 h after the administration of hCG) both eicosanoids declined to their pre-hCG levels. When animals were treated with the cyclooxygenase inhibitor indomethacin at the specific dose of 0.316 mg/rat sc at 1 h before hCG, the ovarian levels of LT B4 and LTs C4/D4/E4 increased to 210% (P less than 0.01) and 113% (P less than 0.05), respectively, above the control levels at 4 h after hCG. Concomitantly, this dosage of indomethacin reduced ovarian prostaglandins (PGs) E and F by 99% (P less than 0.001) and 98% (P less than 0.001), respectively, and it reduced the ovulation rate by 76% (P less than 0.001). Thus it appears this dose of indomethacin blocked the conversion of ovarian arachidonic acid into PGs and shunted this substrate into the lipoxygenase pathways that lead to LT formation. In conclusion, the moderate increase in ovarian LTs is characteristic of inflammatory reactions, and, therefore, these data support the hypothesis that the biochemical events of ovulation resemble an inflammatory process.
Effect of continuous feeding on maternal protein metabolism and fetal growth in the rat
Authors: Ling P. R. , Bistrian B. R. , Blackburn G. L. , Istfan N.
Published: December 2017

Accelerated maternal catabolism ensures adequate fetal growth in the postabsorptive pregnant mammal during late gestation. However, the relative importance of maternal stores and diet in supplying nutrient requirements to the fetus is unknown. We have measured rates of protein synthesis and breakdown in maternal and fetal tissues during continuous intravenous feeding on days 17 and 20 in the rat, using leucine tracer infusions. Nutrient solutions supplying 180 and 250 kcal.kg-1.day-1 were tested. Changes in fetal size were measured in these rats and in rats maintained on ad libitum chow feeding. In comparison to previous results in the postabsorptive rat, leucine flux from endogenous sources into plasma was significantly suppressed, with only 12-20% of circulating leucine derived from maternal tissues. Muscle protein catabolism (previously described on day 20 of gestation) was abolished with continuous feeding. Fetal growth rate averaged 46%/day on day 17 of gestation in all groups but was reduced to 12%/day on day 20 in the rats intravenously fed 180 kcal.kg-1.day-1 and 22%/day in the rats fed 250 kcal.kg-1.day-1. Fetal growth on day 20 in the rats fed rat chow ad libitum was unaffected. We conclude that by decreasing the maternal contribution to circulating nutrients, continuous intravenous feeding may adversely affect fetal growth in late pregnancy. The findings in the current study underline the physiological importance of maternal-fetal nutrient exchange.
Calciotrophic hormone levels and calcium absorption during pregnancy in rats
Authors: Quan-Sheng D. , Miller S. C.
Published: December 2017

The mammalian maternal skeleton stores Ca during pregnancy perhaps for fetal skeletal mineralization in late pregnancy and milk production during lactation. The interrelationships between intestinal Ca absorption and hypertrophy and plasma levels of total Ca, ionized Ca, 25-hydroxyvitamin D3 [25(OH)D], 1,25-dihydroxyvitamin D3 [1,25(OH)2D], and parathyroid hormone (PTH) were determined at different stages of pregnancy in rats. By midpregnancy and before fetal skeletal mineralization, plasma ionized Ca levels, Ca absorption by duodenal tissue in vitro, Ca absorption by the duodenum in situ, and duodenal wet weight were increased and 25(OH)D was decreased. Later in pregnancy, during fetal skeletal mineralization, 1,25(OH)2D and PTH levels were also substantially increased and total serum Ca levels decreased. These data demonstrate changes by midpregnancy, before fetal skeletal mineralization, in maternal mineral homeostasis concomitant with known changes in skeletal metabolism. Some of the early changes in mineral metabolism may occur independent of the vitamin D and PTH endocrine system.
Receptors for IGF-I, but not for IGF-II, on proximal colon epithelial cell apical membranes
Authors: Pillion D. J. , Haskell J. F. , Atchison J. A. , Ganapathy V. , Leibach F. H.
Published: December 2017

Rabbit proximal colon epithelial cell apical membranes, which are known to contain receptors for insulin, were isolated by a Ca2+-precipitation technique. Binding assays with 125I-insulin-like growth factor I (IGF-I) revealed the presence of specific high-affinity binding sites, with 50% inhibition of binding observed at a concentration of 13.7 ng/ml IGF-I. In contrast, 50% inhibition of 125I-IGF-I binding was observed at an insulin concentration of 1.37 micrograms/ml, suggesting that 125I-IGF-I was not binding to insulin receptors present in this tissue. Cross-linking studies revealed an 125I-IGF-I binding subunit of relative molecular weight (Mr) of 130,000 under reducing conditions on docecyl sulfate-polyacrylamide gel electrophoresis that was similar to the IGF-I binding subunit in human placental membranes (Mr 140,000). Binding and cross-linking studies with 125I-insulin-like growth factor II (IGF-II), however, failed to reveal a specific receptor for this peptide in colon epithelial cell membranes. These results establish the coexistence of receptors for IGF-I and insulin, but not IGF-II, on rabbit proximal colon epithelial cell apical membranes and demonstrate that colon epithelial cells are capable of selective synthesis of various peptide hormone receptors.
Sustained increases in plasma epinephrine concentration do not modulate renin release
Authors: Yang H. M. , Lohmeier T. E. , Kivlighn S. D. , Carroll R. G. , Smith M. J.
Published: December 2017

We examined the relationship between plasma renin activity (PRA) and renal arterial pressure (RAP) during 1) control conditions, 2) acute, and 3) chronic intravenous epinephrine (EPI) infusion (125 ng.kg-1.min-1). In eight conscious uninephrectomized dogs maintained on a normal sodium intake, the renin stimulus-response curve (RSRC) was determined by a stepwise reduction in RAP with an inflatable occluder around the renal artery controlled by a servo unit. The RSRC could be approximated by two lines intersecting at a threshold pressure (approximately 20 mmHg below control RAP). In the high-pressure range, PRA was relatively insensitive to changes in RAP, whereas, below threshold pressure, changes in RAP had large effects on PRA. During acute EPI infusion there was approximately a 40% increase in heart rate (control = 57 +/- 3 beats/min) and hematocrit (control = 30 +/- 1%) in association with a rise in plasma EPI concentration from 73 +/- 16 to 1,413 +/- 100 pg/ml; mean arterial pressure (MAP) was unchanged (94 +/- 3 mmHg). Moreover, EPI acutely increased basal PRA from 0.3 +/- 0.1 to 0.8 +/- 0.3 ng angiotensin I.ml-1.h-1 and shifted the RSRC to the right (increasing threshold pressure 7 mmHg) without altering the slope of the RSRC curve either above or below threshold pressure. In contrast, although plasma EPI concentration and hematocrit remained elevated during chronic EPI infusion, heart rate and basal PRA returned to preinfusion values. In addition, there were no significant long-term changes in MAP or in any of the parameters of the RSRC.(ABSTRACT TRUNCATED AT 250 WORDS)
Hormonal and metabolic responses to exercise in humans: effect of sensory nervous blockade
Authors: Kjaer M. , Secher N. H. , Bach F. W. , Sheikh S. , Galbo H.
Published: December 2017

Previous studies have indicated that motor center (“feedforward”) activity is important for hormonal and metabolic responses to exercise. Now, epidural blockade at vertebrae L3-L4 was used to evaluate the importance of afferent neural feedback from working muscles. Six healthy, young males cycled for 20 min at 55 +/- 4% (mean +/- SE) of maximal oxygen uptake with, as well as without, epidural anesthesia. During anesthesia cutaneous sensory blockade was present below segment T11–12, the postexercise ischemic pressor response was attenuated from 34 +/- 9 to 14 +/- 4 mmHg, muscle strength reduced to 80 +/- 5% of control, and perceived exertion (Borg scale) was increased. At rest hormonal and metabolic parameters did not change in response to epidural anesthesia. During exercise, responses of catecholamines, insulin, glucagon, and growth hormone (GH) in plasma as well as glucose production and utilization, plasma free fatty acids, and plasma glycerol were similar in epidural and control experiments (P greater than 0.05). In contrast during submaximal exercise, plasma concentrations of adrenocorticotropin (ACTH) and beta-endorphin increased only in experiments without epidural anesthesia. The data indicate that impulses in afferent nerves from the working muscles are essential for the ACTH and beta-endorphin responses to submaximal dynamic exercise in humans. Afferent nervous activity is probably less important than efferent activity from motor centers for responses of GH, catecholamines and insulin, and, in turn, extramuscular fuel mobilization in exercise.
Epinephrine's effect on metabolic rate is independent of changes in plasma insulin or glucagon
Authors: Staten M. A. , Matthews D. E. , Cryer P. E. , Bier D. M.
Published: December 2017

Epinephrine's effect to increase metabolic rate is accompanied by changes in the plasma concentrations of insulin, glucagon, and metabolic substrates. Because both glucagon and insulin have been reported to affect thermogenesis, these hormones might contribute to or modify the thermogenic response to epinephrine. To determine if the epinephrine-induced increase in metabolic rate is secondary to changes in glucagon or insulin or to changes in the fuels modulated by these hormones, metabolic rate was measured by indirect calorimetry in five normal weight post-absorptive young men on three occasions: study A, an intravenous epinephrine infusion alone; study B, a 4-h “islet clamp” consisting of somatostatin infusion with basal insulin and glucagon replacement; and study C, an intravenous epinephrine infusion combined with the islet clamp. A 1-h base-line period preceded 2 h of epinephrine infusion. During the 4-h islet clamp (study B), metabolic rate and plasma concentrations of epinephrine, insulin, glucagon, and glucose remained unchanged. During the infusion of epinephrine alone (study A), metabolic rate and concentrations of glucagon, free fatty acids, and C-peptide increased as expected. Also as expected, the glycemic response to epinephrine infusion was much larger when insulin and glucagon levels were fixed with the islet clamp (study C). In contrast, the metabolic rate and the free fatty acid concentration responded similarly to epinephrine infusion when insulin and glucagon were fixed (study C) and when they were changing (study A). We conclude that epinephrine increases metabolic rate independently of physiological changes in plasma glucagon or insulin or the circulating fuels they modulate.
Free and sulfoconjugated catecholamine responses to hypoxia in fetal sheep
Authors: Padbury J. F. , Martinez A. M. , Thio S. L. , Burnell E. E. , Humme J. A.
Published: December 2017

Plasma catecholamines circulate either in conjugated or unconjugated forms. In adult humans, sulfoconjugated catecholamines predominate; however, there is considerable variation between species. In a variety of pathophysiological states catecholamine conjugation is believed to represent an important mechanism of inactivation of high circulating catecholamine levels. To date, there have been few data in developing animals or humans on catecholamine sulfoconjugation. We studied the differences in free and sulfoconjugated catecholamines in full term (141 +/- 1 days) and preterm (123 +/- 1 days) chronically catheterized fetal sheep and determined the changes in free and sulfoconjugated catecholamines in response to hypoxia. The results demonstrate that term and preterm animals have a comparable percentage of basal circulating sulfoconjugated catecholamines (free-to-total ratio 50-60%). In response to hypoxia, both free and sulfoconjugated catecholamines were promptly elevated with significant increases in each by 5 min of hypoxia. This was true for both term and pretern animals. The proportion of free and total catecholamines remained relatively constant during hypoxia despite a 5- to 10-fold increase in circulating levels of each. These data demonstrate that fetal sheep, as early as 80% gestation, have a well developed mechanism for sulfoconjugation and subsequent inactivation of the high circulating levels of catecholamines seen during fetal and newborn life.
Arachidonic acid affects membrane ionic conductances of GH3 pituitary cells
Authors: Vacher P. , McKenzie J. , Dufy B.
Published: December 2017

Arachidonic acid (AA) stimulates prolactin release from pituitary cells, by mechanisms not yet understood. In this work, we analyzed the effects of AA on membrane ionic conductances in a clonal line of anterior pituitary cells (GH3/B6), finding time- and dose-dependent effects of AA on their membrane ionic conductances. The predominant response at concentrations between 100 nM and 10 microM was a prolongation of the action potential (AP) and an increase in the transient after-hyperpolarization potential. Voltage clamp studies showed that this was associated with a decrease in a voltage-dependent potassium current and an increase in a voltage-dependent calcium current. In some cells (30%) the effect of AP duration was less important, but spike firing was enhanced. For the highest concentrations used (1 and 10 microM) the effects described above were preceded by hyperpolarization of the cell membrane; in voltage clamp it was shown that this hyperpolarization resulted from the activation of a calcium-dependent potassium conductance suspected to be due to the release of intracellular calcium. The calcium store affected by AA was, at least in part, insensitive to vanadate and heparin. These data suggest that AA may enhance intracellular calcium concentration by increasing calcium entry during each voltage-dependent calcium AP, by increasing the spike frequency, or by releasing calcium from an intracellular compartment. The resulting rise in cytosolic free calcium concentration may be a key link in the process by which AA stimulates prolactin release in GH3/B6 pituitary cells.
Effect of loss of first-phase insulin secretion on hepatic glucose production and tissue glucose disposal in humans
Authors: Luzi L. , DeFronzo R. A.
Published: December 2017

To examine the importance of first-phase insulin secretion on total body glucose homeostasis, six normal subjects (age, 24 +/- 1 yr; ideal body wt, 100 +/- 1%) received three hyperglycemic (+75 mg/100 ml) clamp studies in combination with [3-3H]glucose: study I, 150 min hyperglycemic clamp; study II, hyperglycemic clamp plus somatostatin (6 micrograms/min) plus basal glucagon replacement (0.4 ng.kg-1.min-1) plus an insulin infusion designed to mimic only the second phase of insulin secretion; and study III, hyperglycemic clamp plus somatostatin plus basal glucagon plus an insulin infusion designed to mimic both the first and second phase of insulin secretion. Basal plasma C-peptide concentrations averaged 0.21 +/- 0.01 pmol/ml in the three study protocols. In study I the plasma C-peptide response demonstrated an early burst within the first 10 min followed by a gradually increasing phase of C-peptide secretion that lasted until the end of the study. In studies II and III plasma C-peptide declined within the first 10 min after somatostatin was started and averaged 0.06 +/- 0.01 and 0.05 +/- 0.01 pmol/min, respectively. Basal hepatic glucose production (2.3 +/- 0.2 mg.kg-1.min-1) was suppressed by 90% at 20 min and remained suppressed thereafter in studies I and III. In contrast, in study II hepatic glucose production was inhibited by only 50% (1.1 +/- 0.2 mg.kg-1.min-1) at 60 min (P less than 0.01 vs. studies I and III) and remained incompletely suppressed even after 150 min.(ABSTRACT TRUNCATED AT 250 WORDS)
1,25-Dihydroxyvitamin D3-induced alterations of lipid metabolism in human monocyte-macrophages
Authors: Roullet J. B. , Haluska M. , Morchoisne O. , McCarron D. A.
Published: December 2017

An in vivo atherogenic role of dietary vitamin D has been postulated. To address this hypothesis we sought to determine the in vitro effects of its active circulating metabolite, 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] on lipid metabolism in human monocyte-derived macrophages. When cultured 6 days in the presence of 10(-8) M 1,25(OH)2D3 monocyte-macrophages accumulated significantly more triglycerides than control cells: 987.6 +/- 26.8 vs. 779.3 +/- 24.1 micrograms/mg protein (P less than 0.001). Triglyceride accumulation was associated with a hormone-induced stimulation of triglyceride synthesis as determined by [3H]oleate incorporation into cellular triglycerides. The effect of the hormone was significant after 24 h and dose dependent [10(-11) to 10(-8) M 1,25(OH)2D3]. It was specific since 10(-7) M 25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 did not stimulate triglyceride synthesis, and its magnitude decreased from 1 to 9 days of culture. 1,25(OH)2D3 (10(-8) M) modified the cholesteryl ester metabolism of monocyte-macrophages only in the presence of acetylated low-density lipoproteins (50 micrograms/ml); it induced a significant increase of cellular cholesteryl ester content (21.9 +/- 1.1 vs. 11.7 +/- 1.7 micrograms/mg protein; P less than 0.001) and of esterification rate of cholesterol measured by [3H]oleate incorporation into cellular cholesteryl esters (17.2 +/- 0.9 vs. 6.5 +/- 0.3 nmol.mg protein-1.24 h-1; P less than 0.001) by comparison with control cells. These results show that 1,25(OH)2D3 alters in vitro lipid metabolism in the human monocyte-macrophage and suggest a new in vivo role for the hormone.
Metabolic and sympathetic nerve activities of brown adipose tissue in tube-fed rats
Authors: Saito M. , Minokoshi Y. , Shimazu T.
Published: December 2017

To investigate possible contribution of oropharyngeal factors to thermogenesis in brown adipose tissue (BAT), rats were given a liquid diet orally or through a gastric tube for 4-12 days. Tube-fed rats weighed more and had heavier white adipose tissue than orally fed rats, although they were given the same amount of the diet. Triglyceride content of the interscapular BAT was higher in tube-fed rats, but the rate of fatty acid synthesis was lower than those in orally fed rats. Uptake of 2-deoxyglucose (2DG), an index of glucose utilization, in BAT was also lower in tube-fed rats. Surgical sympathetic denervation of BAT reduced both lipogenic activity and 2DG uptake to abolish the differences in these metabolic activities between tube-fed and orally fed rats. Sympathetic nerve activity in the BAT assessed from the turnover rate of norepinephrine in the tissue was lower in tube-fed rats. The results suggest that there is a sympathetically mediated mechanism dependent on afferent input from oropharyngeal receptors to regulate BAT thermogenesis and that greater fattening seen after tube feeding may be attributable to a decreased BAT thermogenesis because of the absence of food-induced sensory stimulation.
Abnormal insulin metabolism by specific organs from rats with spontaneous hypertension
Authors: Mondon C. E. , Reaven G. M. , Azhar S. , Lee C. M. , Rabkin R.
Published: December 2017

Spontaneously hypertensive rats (SHR) have been shown to be both insulin resistant and hyperinsulinemic after oral glucose administration or infusion of exogenous insulin during an insulin suppression test. To determine if this hyperinsulinemia may be due to decreased removal of insulin, the metabolic clearance (k) of insulin was measured in isolated perfused liver, kidney, and hindlimb skeletal muscle from SHR and Wistar-Kyoto (WKY) control rats. The data indicate that the k for insulin removal by liver was similar in SHR and WKY rats, averaging 287 +/- 18 and 271 +/- 10 microliters.min-1.g-1 liver, respectively. In contrast, the k for insulin removal by hindlimbs from SHR was decreased 37% (P less than 0.001) compared with WKY rats (8.6 +/- 0.5 vs. 13.7 +/- 0.7 microliters.min-1.g-1 muscle), and this decrease was not accompanied by decreased binding of insulin to its receptor in plantaris muscle. Although the removal of insulin by glomerular filtration was similar in SHR and WKY rats (653 +/- 64 microliters/min vs. 665 +/- 90 microliters.min-1.kidney-1), total insulin removal by kidney was significantly lower (P less than 0.05) in SHR (710 +/- 78 microliters/min) compared with WKY rats (962 +/- 67 microliters/min), due to decreased peritubular clearance of insulin in SHR (56 +/- 73 vs. 297 +/- 59 microliters/min, P less than 0.05). These findings suggest that the decreased clearance of insulin in SHR rats was possibly not due to impaired hepatic removal of insulin but rather to decreased removal by skeletal muscle and kidneys.(ABSTRACT TRUNCATED AT 250 WORDS)
Downregulation of beta-adrenergic receptor-mediated function during sodium restriction in humans
Authors: Rosen S. G. , Linares O. A. , Smith M. J. , Halter J. B.
Published: December 2017

Mononuclear leukocyte (MNL) beta 2-adrenergic receptor (beta 2-AR) binding and its linked adenylate cyclase sensitivity to isoproterenol were measured in nine healthy humans prior to and after 7 days of dietary sodium restriction to determine whether chronic physiological increases in plasma norepinephrine (NE) are associated with the downregulation of beta-AR-mediated function. Sodium restriction resulted in an increase in the plasma NE concentration (P less than 0.02) and decreases in MNL beta 2-AR density (P less than 0.001), affinity for antagonist (P less than 0.001), and adenylate cyclase sensitivity to isoproterenol (ANOVA, P less than 0.01). To determine whether this downregulation of MNL beta 2-AR-mediated function is related to the increased plasma NE concentration or to increased extravascular NE release, NE kinetics was assessed using compartmental analysis in each subject prior to and after sodium restriction. Sodium restriction caused a decrease in the plasma NE metabolic clearance rate (P less than 0.005) and in the volume of distribution of NE in the intravascular compartment (P less than 0.005), whereas the extravascular NE release rate was unchanged. Our data suggest that the downregulation of MNL beta 2-AR-mediated function in humans during dietary sodium restriction is a response to the increase in plasma NE.
Diabetes decreases creatine kinase enzyme activity and mRNA level in the rat heart
Authors: Popovich B. K. , Boheler K. R. , Dillmann W. H.
Published: December 2017

Several of the adenosinetriphosphatase enzymes that are responsible for cardiac muscle contraction rely on high-energy phosphates supplied by the creatine kinase (CK) system. Experimental diabetes mellitus has been shown to cause a decrease in the maximal contractile performance of the heart. We postulated that the decrease in contractile performance may be explained in part by a decrease in CK enzyme activity. To evaluate this possibility, we determined the level of CK activity and isoenzyme distribution in ventricular homogenates from normal, diabetic, and insulin-treated diabetic rats. We found that total CK activity was decreased by 35% in diabetic hearts and that a 66% reduction in the cardiac-specific MB isoenzyme occurs. Using a cDNA probe for CK-muscle (M) RNA in Northern blot analysis, we determined that a 61.1% decrease in CK-M mRNA occurs in diabetes. Chronic insulin therapy for 1 mo restores CK-M mRNA levels and enzyme activity. In conclusion, diabetes-induced CK enzyme decreases are mediated in part by a lower level of CK-M mRNA that codes for the major CK-M subunit protein. Decreased performance of the CK system may contribute to diabetic cardiomyopathy.
Whole body protein synthesis: studies with different amino acids in the rat
Authors: Obled C. , Barre F. , Millward D. J. , Arnal M.
Published: December 2017

These studies were undertaken to determine to what extent constant infusion measurements and plasma sampling could provide sensible answers for rates of whole body protein turnover and also which amino acid would be the most representative probe of whole body protein turnover. Whole body protein synthesis rates were estimated in 70-g rats with L-[U-14C]threonine, L-[U-14C]lysine, L-[U-14C]tyrosine, L-[U-14C]phenylalanine, and L-[1-14C]leucine by either simultaneous tracer infusion of four amino acids or by injections of large quantities of 14C-labeled amino acids. In the infusion experiment, indirect estimates of whole body protein turnover based on free amino acid specific radioactivity and stochastic modeling were compared with direct measurement of the incorporation of the tracer into proteins. These two methods of analysis provided similar results for each amino acid, although in each case fractional synthesis rates were lower (by between 26 and 63%) when calculations were based on plasma rather than tissue specific radioactivity. With the flooding-dose method, whole body fractional protein synthesis rates were 41.4, 25.6, 31.1, and 31.4% with threonine, lysine, phenylalanine, and leucine, respectively. These values were similar to those obtained by the continuous infusion method using tissue specific radioactivity for threonine and lysine. For leucine, however, the flooding-dose method provided an intermediate value between the two estimates derived either from the plasma or the tissue specific radioactivity in the infusion method.(ABSTRACT TRUNCATED AT 250 WORDS)