bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies of gut and hepatic metabolism in conscious rabbits
Authors: Pere M. C. , Gilbert M. , Assan R. , Battaglia F. C.
Published: December 2017

The present study was designed to develop the techniques for chronic catheterization of the hepatic and portal venous circulation in conscious rabbits and to apply these techniques to a study of hepatic metabolism in this species. Experiments were made after an 18-h fast and for 4 h after the initial feeding. Measurements of arteriovenous differences of substrates were combined with measurements of hepatic and gastrointestinal blood flow. Hepatic glucose production was suppressed by 60% at 1 h and had returned to control levels by 4 h. The hepatic uptake of lactate declined slightly at 1 h and had returned to control level 2 h after the meal. There was a marked and rapid fall in hepatic ketone body output after refeeding. Although amino acid concentrations displayed a transient increase 1 h after the meal, only the arterial concentration of branched-chain amino acids remained significantly elevated for 4 h. The total hepatic uptake of the gluconeogenic amino acids (alanine, serine, threonine) remained constant. Refeeding resulted in a doubling of arterial insulin concentrations at 1 h followed by a progressive decline over the next 3 h. It is concluded that in rabbits fed a mixed meal partial suppression of hepatic glucose output is mainly due to a decline in glycogenolysis rather than a decrease in gluconeogenesis, shortly after refeeding the liver is able to virtually shut off its ketone body production, the major gluconeogenic precursors (lactate, alanine, glycine, serine, and threonine) may contribute to approximately 40% of the glucose release.(ABSTRACT TRUNCATED AT 250 WORDS)
Static mechanical properties of lungs from adrenalectomized pneumonectomized rats
Authors: Bennett R. A. , Addison J. L. , Rannels D. E.
Published: December 2017

The pressure-volume characteristics of saline-filled lungs of adult rats were studied after left pneumonectomy (PNX), with or without prior bilateral adrenalectomy (ADX). Male Sprague-Dawley rats (200–240 g) remained unoperated (UNOP) or were divided into three surgical groups: left PNX, ADX alone, or PNX preceded 5 days earlier by ADX (ADX/PNX). Two weeks post-PNX, maximum lung volumes [MLV; measured at transpulmonary pressure (Ptp) of 10 cmH2O] of right lungs from PNX and ADX/PNX rats remained 20 and 37% below UNOP values, respectively, although right lung mass was equal to or exceeded that of both lungs in UNOP controls. Four weeks postoperative, MLVs approached but remained below normal values. In the ADX/PNX group, MLV was 35% below that predicted for lung mass. The effects of ADX on both lung mass and MLV were blocked by daily injections of ADX/PNX animals with hydrocortisone acetate (5 mg/kg). Neither PNX nor ADX/PNX altered lung volume at zero Ptp or changed the concentration of lung collagen or elastin. When corrected for differences in MLV, relative tissue compliance was unaffected by PNX but was reduced by ADX/PNX. These results indicate that MLV is not restored in parallel with lung mass during the rapid phase of compensatory tissue growth, which follows partial pneumonectomy, and that this discrepancy is accentuated by adrenalectomy.
Study of ketone body kinetics in children by a combined perfusion of 13C and 2H3 tracers
Authors: Bougneres P. F. , Ferre P.
Published: December 2017

Ketone body kinetics were quantified in six children (3-5 yr old), who were fasted for 13-22 h, by a combined perfusion of [3-13C]acetoacetate ([13C]AcAc) and D-(-)-beta-[4,4,4-2H3]hydroxybutyrate (beta-[2H3]OHB) and gas chromatography-mass spectrometry analysis. Results were analyzed according to the "single-pool" (combined enrichments) or the "two-accessible pools" models. After 20-22 h of fasting, ketone body turnover rate was 30-50 mumol.kg-1.min-1, a rate achieved after several days of fasting in adults. At low ketosis, acetoacetate was the ketone body preferentially synthesized de novo and utilized irreversibly. When ketosis increased, acetoacetate irreversible disposal was not enhanced, since it was largely converted into beta-OHB, whereas beta-OHB irreversible disposal was very much increased. The single-pool and two-pool models gave similar ketone body turnover rates when [13C]AcAc was the tracer, whereas the use of beta-[2H3]OHB gave some more divergent results, especially at low ketosis. These studies demonstrate that ketogenesis is very active in short-term fasted children and that the use of a combined infusion of [13C]AcAc and beta-[2H3]OHB is a convenient way to give insight into individual ketone body kinetics.
Models to interpret kinetic data in stable isotope tracer studies
Authors: Cobelli C. , Toffolo G. , Bier D. M. , Nosadini R.
Published: December 2017

In contrast to "weightless" radioactive tracers, stable isotope tracers have nonnegligible mass and are naturally present in the system, and the measured variable is a ratio of two isotopic species. These features do not allow stable isotopic tracer data analysis using straightforward analogy with radioactive tracer approaches, even though this practice is common. In this study, we present kinetic variables, models, and measurements for the analysis and interpretation of stable isotope tracer data. Assumptions and mathematical techniques for modeling the data when perturbation is both nonnegligible and negligible are discussed. Emphasis is placed on the rich information content of the dynamic portion of a stable isotope tracer curve and on the role of compartmental and noncompartmental modeling approaches for its interpretation. A presumed and commonly used analogy between the radioactive specific activity and stable isotopic enrichment is shown to be incorrect. We show that the proper analogue of specific activity is the tracer-to-tracee molar ratio. This variable is not a directly measurable one, but a formula is derived that allows its computation from the data. A method for reconstructing the time course in blood of the concentration component due to endogenous synthesis is presented. This allows measurement of the extent of the perturbation in the case where a nonweightless tracer is used. Special attention is given to data analysis originating from a multiple tracer experiment, a configuration necessary for studying more complex systems, e.g., the kinetics of interacting substrates.
Comparison of [14C]glucose and [14C]deoxyglucose as tracers of brain glucose use
Authors: Hawkins R. A. , Mans A. M. , Davis D. W. , DeJoseph M. R.
Published: December 2017

Because glucose metabolism and functional activity in brain regions are normally coupled, knowledge of regional brain glucose use can yield insights into regional functional activity. The deoxyglucose (DG) method is widely used for this purpose in experimental animals and humans but questions have arisen regarding its limits and accuracy. Therefore an experiment was designed to compare the DG method on a structure-by-structure basis with another tracer of glucose use, [6-14C]glucose, in normal rats. The cerebral metabolic rates obtained using the two tracers were similar in the telencephalon, but the results using DG were substantially lower in the midbrain and hindbrain (diencephalon, 18%; mesencephalon, 20%; metencephalon, 29%; and myelencephalon, 35%). The primary DG metabolite, DG 6-phosphate (DG-6-P) was found to disappear in a non-uniform manner from the major brain structures: telencephalon less than diencephalon less than mesencephalon = metencephalon less than myelencephalon. Thus a correlation was found between the rate of DG-6-P loss and the extent to which the DG method gave lower values of glucose use. Thus this may explain, at least in part, the discrepancies between the two methods.
Ouabain induces secretion of proatrial natriuretic factor by rat atrial cardiocytes
Authors: Bloch K. D. , Zamir N. , Lichtstein D. , Seidman C. E. , Seidman J. G.
Published: December 2017

Neonatal rat atrial and ventricular cardiocytes in primary culture secrete proatrial natriuretic factor (proANF), the 126-amino acid precursor of atrial natriuretic factor (ANF). The cellular mechanisms of proANF secretion differ in the two cell types: atrial cardiocytes store proANF in abundant secretory granules before secretion, whereas ventricular cardiocytes secrete the precursor rapidly after synthesis. In this study, we used the Na+-K+-adenosinetriphosphatase (ATPase) inhibitor ouabain to investigate the functional significance of these differing secretory mechanisms. Ouabain increased immunoreactive ANF (iANF) secretion by atrial cardiocytes 1.5- to 2.4-fold. Metabolic labeling studies demonstrated that proANF was the form of iANF released in response to ouabain. Ventricular secretion of iANF was unaffected by the Na+-K+-ATPase inhibitor. These observations are consistent with a model in which atrial cardiocytes are able to release proANF via a regulated secretory pathway, whereas ventricular cardiocytes utilize a constitutive secretory pathway. The ability of ouabain to stimulate proANF secretion suggests that an increase in intracellular calcium concentration may promote fusion of secretory granules with atrial cell membranes thereby mediating exocytosis of stored proANF.
Effect of thyroidectomy on rat T-kininogen
Authors: Bouhnik J. , Savoie F. , Baussant T. , Michaud A. , Alhenc-Gelas F. , Corvol P.
Published: December 2017

The thyroid state affects the inflammatory reaction; in general, thyroid hormones increase and hypothyroidism decreases the inflammatory response. T-kininogen is an acute phase protein that inhibits cysteine proteases. The influence of thyroidectomy and 3,5,3'-triiodothyronine (T3)-treatment of thyroidectomized animals was examined on rat plasma T-kininogen concentration. Thyroidectomy increased the T-kininogen level three- to fourfold. A single administration of 10 micrograms T3/100 g body wt in 7-wk postthyroidectomy rats decreased it by approximately 50%. Daily treatment with 0.5 micrograms T3/100 g body wt prevented the increases in T-kininogen. The modifications in plasma T-kininogen concentration were confirmed by high-performance liquid chromatography analysis of the plasma kinins released by trypsin. They demonstrated that T-kinin was greatly and bradykinin slightly increased after thyroidectomy and that T-kinin concentration in T3-treated thyroidectomized animals was almost restored to the control level. The ability of thyroidectomized animals to produce a higher plasma T-kininogen compared with euthyroid animals was further documented when T-kininogen production was stimulated by laparotomy. This procedure resulted in a fourfold increase in T-kininogen in thyroidectomized animals compared with controls. The increase of plasma T-kininogen level after thyroidectomy results from a more active synthesis of the protein, since liver slices from thyroidectomized rats synthesized T-kininogen at a rate about two times higher than normal rats. These data may account at least in part for the diminution of the inflammatory response observed after thyroidectomy.
Relative contributions of the nervous system and hormones to CNS-mediated hyperglycemia
Authors: Iguchi A. , Gotoh M. , Matsunaga H. , Yatomi A. , Honmura A. , Yanase M. , Sakamoto N.
Published: December 2017

We quantitatively determined the relative contributions of hormonal factors and the nervous system to the total glucose response after stimulation of the cholinergic neurons in the central nervous system of fed rats. Hepatic venous plasma glucose, glucagon, insulin, epinephrine, and norepinephrine were measured during 120 min after injection of neostigmine (5 X 10(-8) mol) into the third cerebral ventricle in rats subjected to bilateral adrenodemedullation (ADMX) to prevent epinephrine secretion (observed insulin secretion), with and without intravenous infusion of somatostatin to prevent glucagon and insulin secretion. Injection of neostigmine in intact rats resulted in increases in glucose, glucagon, epinephrine, and norepinephrine. Comparison of glucose areas suggests that 22% of the hyperglycemic response is due to the glucagon effect, that 29% is due to the epinephrine effect, and that an unknown factor other than epinephrine or glucagon, which may include activation through direct neural innervation of the liver via alpha-adrenergic receptor, contributes 49%. The suppressive effect of epinephrine on insulin secretion, which is potentially stimulated by direct neural activation of the pancreas, contributes 18% of the net hyperglycemia.
Absorption of epidermal growth factor and insulin in rabbit renal proximal tubules
Authors: Nielsen S. , Nexo E. , Christensen E. I.
Published: December 2017

Epidermal growth factor (EGF), which stimulates the growth of a variety of tissues, was originally isolated from mouse submandibular glands. Human EGF (hEGF) has been isolated from the urine, and cDNA encoding for EGF has been isolated from the human kidney. Thus the kidney may represent an alternate source of EGF. Another potential explanation for the urinary content of EGF is a limited reabsorption of filtered EGF and/or a transtubular transport of EGF from the peritubular compartment to the lumen. Therefore, we exposed isolated and perfused rabbit proximal tubules to 125I-hEGF either in the perfusate or in the bath fluid. Luminal uptake of hEGF was compared with uptake of 125I-porcine insulin, which is known to be taken up with large efficiency. The results demonstrate that only 4% of the perfused 125I-hEGF load was taken up per millimeter of tubule length compared with 73% for insulin. Furthermore, hEGF is subject to a small transtubular transport from bath to lumen. Renal clearance of endogenous rabbit EGF was also measured and was comparable with that of creatinine. Thus in conclusion this study strongly suggests that filtered EGF to a large extent remains in the ultrafiltrate, unlike insulin, which is removed within the first few millimeters of proximal tubule. In addition filtered EGF in preurine is probably supplied by EGF transported intact across the proximal tubular epithelium.
Oxygen consumption after massive sympathetic nervous system discharge
Authors: Lang S. A. , Maron M. B. , Signs S. A.
Published: December 2017

We evaluated the possibility that massive, sympathetic nervous system (SNS) activation [as may precede the development of neurogenic pulmonary edema (NPE)] increases O2 demand. O2 consumption (VO2) and plasma concentrations of the calorigenic agents, epinephrine (EPI) and norepinephrine (NE) were measured in alpha-chloralose-anesthetized dogs under control conditions and for 3 h after the administration of either 1) intracisternal (ic) veratrine to activate the SNS, 2) intravenous (iv) veratrine, 3) ic saline, or 4) ic veratrine, after clamping the adrenal blood vessels. VO2 increased 31.7 +/- 3.6% (SE), and EPI and NE increased to, respectively, 30,853 +/- 8,347 and 8,176 +/- 2,104 pg/ml in the ic veratrine group. No increases in VO2 and EPI and attenuated increases in NE were observed in the ic veratrine animals with clamped adrenals. No significant increases in VO2 or catecholamine concentrations were observed after ic saline or iv veratrine administration. These data suggest that the elevated VO2 may have been mediated by adrenal catecholamines and that an increased metabolic rate may complicate the ability of patients with severe NPE to balance O2 supply with demand.
Direct effect of CNS on insulin hypersecretion in obese Zucker rats: involvement of vagus nerve
Authors: Lee H. C. , Curry D. L. , Stern J. S.
Published: December 2017

It is hypothesized that the vagus nerve makes a major contribution to pancreatic insulin hypersecretion in the genetically obese rat (fa/fa) via direct pancreatic innervation. An in situ brain-pancreas perfusion model with intact pancreatic central nervous system (CNS) innervation was used in these studies. The dynamics of insulin secretion in response to a 40-min glucose stimulus (200 mg/dl) was investigated in CNS intact (INT), bilateral cervical vagotomized (VGX), and CNS functionally ablated (ABL) 11- to 12-wk-old homozygous lean (Fa/Fa) and obese (fa/fa) female Zucker rats. The overall pattern of insulin secretory dynamics from obese and lean rats was similar. However, insulin released during the entire 40-min perfusion period by pancreata from obese rats was significantly greater than in lean rats. In lean rats, there was no significant difference in insulin secretion from pancreata of CNS-INT, VGX, and ABL rats. In obese rats, CNS-INT pancreata secreted almost twice as much insulin as pancreata from obese ABL rats and four times as much insulin as CNS-INT lean rats. This demonstrates that hypersecretion of insulin in obese Zucker rats is comprised of a significant direct CNS component. Although vagotomy had little effect on CNS-INT lean rats, it reversed the CNS component of hypersecretion present in CNS-INT obese rats. Because insulin secretion in CNS-INT obese rats was lowered by vagotomy to that equivalent to values of CNS-ABL obese rats, this demonstrates a significant contribution by the parasympathetic nervous system to the hyperinsulinemia seen in the Zucker obese rat that is attributed to direct parasympathetic innervation of the pancreas.
Absorption of plasma proteins from peritoneal cavity of normal rats
Authors: Regoeczi E. , Zaimi O. , Chindemi P. A. , Charlwood P. A.
Published: December 2017

The present study was undertaken to examine whether the uptake of plasma proteins from the peritoneal cavity is quantitative so that tracers could be introduced that way for measuring their turnover. To this end, the metabolic behavior of seven homologous plasma proteins, labeled with 125I, was compared in rats after intravenous or intraperitoneal administration. The animals were maintained under physiological conditions. Total body radiation measurements showed that the degradation rates of albumin, immunoglobulins A and G, alpha 1-macroglobulin, and transferrin were the same regardless of the route of injection. This implies that these proteins are quantitatively absorbed from the peritoneum without undergoing modifications. The half-life of intraperitoneally injected alpha 1-acid glycoprotein was consistently shorter by an average 9%, thus suggesting that this protein becomes slightly altered if introduced that way. Only one-half of intraperitoneally injected fibrinogen survived normally, whereas the other underwent rapid degradation. The surviving molecules had the same half-life as fibrinogen injected intravenously. The fraction of surviving fibrinogen could be augmented by mixing the dose with serum. Within a wide range of concentrations and quantities injected, the degradation rate of transferrin remained the same. Analysis by deconvolution of the plasma curves of albumin and alpha 1-macroglobulin absorbed from the peritoneum showed that the transport process was independent of protein size and, at least up to 35 mg, of the amount injected. According to the same technique, intraperitoneally administered diferric transferrin retained its iron during passage into the circulation.
Escape of hepatic glucose production during hyperglycemic clamp
Authors: Elahi D. , Meneilly G. S. , Minaker K. L. , Andersen D. K. , Rowe J. W.
Published: December 2017

The role of the pattern of insulin secretion on hepatic glucose production (HGP) was evaluated with hyperglycemic and euglycemic clamp studies in six normal young nonobese subjects. In the hyperglycemic studies, glucose levels were raised and maintained at 98 mg/dl above basal for 150 min. Each subject responded with a biphasic pattern of immunoreactive insulin (IRI) release. HGP was completely suppressed by 20 min, coincident with the first-phase insulin release. HGP then rose steadily, surpassing the basal rate by 100 min when IRI had reached the peak levels of the first phase. By 130 min, when IRI had surpassed the peak first-phase levels, HGP began to fall. In the euglycemic studies with a square wave of hyperinsulinemia (approximately 25 microU/ml), HGP was suppressed to approximately 60% of basal and remained at that rate. We next repeated the hyperglycemic studies with somatostatin, glucagon, and insulin infusions. In these studies with a square wave of hyperinsulinemia (approximately 40 microU/ml, the level observed during the first phase IRI of the previous hyperglycemic clamps), HGP was suppressed to approximately 43% of basal rate and remained at that rate. These studies indicate insulin regulation of HGP is not only dependent on insulin level but may be strongly influenced by the pattern, over time, of insulin secretion.
Synthesis and characterization of fluorescein- and rhodamine-labeled probes for vasotocin receptors
Authors: Buku A. , Masur S. , Eggena P.
Published: December 2017

Fluorescent analogues of vasotocin, [1-(beta-mercaptopropionic acid), 9-(p-aminofluoresceinylphenylalanine)]arginine vasotocin [[MPA1, (p-NH2flu)Phe9]AVT] and [1-(beta'-mercaptopropionic acid), 9-(p-amino rhodaminylphenylalanine)]AVT [[MPA1, (p-NH2rhod)-Phe9]AVT], were synthesized by the solid-phase method. These compounds yielded half-maximal hydrosmotic responses (half-maximal values) in the toad urinary bladder at 2 X 10(-9) M. Epifluorescence microscopy showed punctate basal localization of analogues on the majority of bladder epithelial cells within 20 min. The cellular localization was prevented by vasotocin. These fluorescent analogues may serve as useful probes for vasotocin receptors in toad bladder and in tissues from other species that use vasotocin as their antidiuretic-pressor hormone.
Impaired aldosterone production by long-term infusion of atrial natriuretic factor
Authors: Nagano M. , Bravo E. L.
Published: December 2017

This study assessed the effect of chronic infusions of atrial natriuretic factor (ANF) on in vivo and in vitro production of aldosterone. Vehicle (saline) or rat ANF-(99-126) was intravenously infused at 100 ng.kg-1.h-1 for 5 consecutive days into male New Zealand White rabbits. At 5 days plasma ANF was 18 +/- 4.1 pg/ml in vehicle-infused and 48.5 +/- 9.0 in ANF-infused rabbits (P less than 0.01). Plasma renin activity was significantly less in ANF-infused rabbits (2.99 +/- 0.35 vs. 0.77 +/- 0.12 ng.ml-1.h-1, P less than 0.01); however no differences were observed in the basal plasma concentrations of aldosterone, corticosterone, potassium, or hematocrit. In in vivo studies, chronically administered ANF attenuated plasma aldosterone, but not pressor, responses to acutely infused angiotensin II given at doses of 4, 16, and 64 ng.kg-1.min-1 for 20 min each. In in vitro experiments, collagenase-dispersed adrenal capsular cells from ANF-infused rabbits exhibited significantly reduced maximal responses to adrenocorticotropic hormone, angiotensin II, and potassium. These results suggest that chronic small increases in circulating ANF can blunt selectively adrenocortical responses to aldosterone secretagogues without affecting pressor responses to angiotensin II.
Energy expenditure in humans: effects of dietary fat and carbohydrate
Authors: Abbott W. G. , Howard B. V. , Ruotolo G. , Ravussin E.
Published: December 2017

A high-dietary fat intake may be an important environmental factor leading to obesity in some people. The mechanism could be either a decrease in energy expenditure and/or an increase in caloric intake. To determine the relative importance of these mechanisms we measured 24-h energy expenditure in a whole body calorimeter in 14 nondiabetic subjects and in six subjects with non-insulin-dependent diabetes mellitus, eating isocaloric, weight-maintenance, high-fat, and high-carbohydrate diets. All subjects were Pima Indians. In nondiabetics, the mean total 24-h energy expenditure was similar (2,436 +/- 103 vs. 2,359 +/- 82 kcal/day) on high-fat and high-carbohydrate diets, respectively. The means for sleeping and resting metabolic rates, thermic effect of food, and spontaneous physical activity were unchanged. Similar results were obtained in the diabetic subjects. In summary, using a whole body calorimeter, we found no evidence of a decrease in 24-h energy expenditure on a high-fat diet compared with a high-carbohydrate diet.
ICV infusion of corticosterone antagonizes ICV-aldosterone hypertension
Authors: Gomez-Sanchez E. P. , Venkataraman M. T. , Thwaites D. , Fort C.
Published: December 2017

There is evidence of crucial central nervous system involvement in the pathogenesis of mineralocorticoid-excess salt hypertension, as well as data indicating that corticosterone is the predominant ligand for the type I adrenocorticoid receptor in the brain. Miniosmotic pumps were used to deliver artificial cerebrospinal fluid (CSF), aldosterone (10 ng/h), corticosterone (10 or 20 ng/h), aldosterone (10 ng/h) plus corticosterone [10 ng/h intracerebroventricularly (icv)], or aldosterone (10 ng/h) plus corticosterone (20 ng/h icv). All animals were sensitized to mineralocorticoid hypertension by removing the right kidney and offering saline to drink. Indirect blood pressure by the unheated tail-cuff method and weights were measured twice weekly; 24-h urine volumes were measured once a week. The blood pressures of the four groups did not differ statistically before infusion. The blood pressures of those animals receiving CSF or corticosterone were not significantly elevated after 4-5 wk of intracerebroventricular infusion, whereas the aldosterone group had become significantly elevated within 2 wk. A similar study was done comparing the effects of intracerebroventricular infusion of aldosterone (10 ng/h), aldosterone (10 ng/h) and RU26988 (20 ng/h), and RU26988 (20 ng/h). RU26988, a selective type II receptor agonist, had no effect on the blood pressure, nor did it alter the pressor effect of intracerebroventricular aldosterone. The concomitant infusion of corticosterone antagonized the increase in blood pressure in a dose-dependent manner. Neither steroid nor their combinations produced significant differences in daily urine volume or body weight gain compared with the CSF group.
Effects of extracellular pH, CO2, and HCO3- on ketogenesis in perfused rat liver
Authors: Wu G. Y. , Gunasekara A. , Brunengraber H. , Marliss E. B.
Published: December 2017

Effects of extracellular pH, CO2, and HCO3- on ketone body production from octanoate were studied in perfused livers from fasted rats. pH was adjusted to 7.1-7.5 by varying perfusate [HCO3-] and [CO2], where brackets denote concentration. At constant 25 mM [HCO3-], total production of beta-hydroxybutyrate (beta-OHB) + acetoacetate (AcAc) was constant from pH 7.1 to 7.5. However, the [beta-OHB]/[AcAc] ratio decreased from 1.60 to 1.00 when pH decreased from 7.3 to 7.1; there was no change at pH 7.4. At constant [CO2], decreasing pH from 7.4 to 7.1 did not alter either total ketogenesis or the [beta-OHB]/[AcAc] ratio. This suggests that high [CO2] rather than low pH was responsible for the alteration in the redox ratio. At constant pH of 7.4, variations in [HCO3-] between 15 and 25 mM did not influence total ketogenesis or the [beta-OHB]/[AcAc] ratio. However, increasing [HCO3-] from 25 to 35 mM decreased the [beta-OHB]/[AcAc] ratio from 1.76 to approximately 1, again without affecting total ketogenesis. At constant 1.75 mM [CO2], increasing [HCO3-] from 25 to 35 mM also reduced the [beta-OHB]/[AcAc] ratio from 1.63 to approximately 1, suggesting that the effect of high [HCO3-] on this redox ratio can be ascribed to HCO3- itself. It is concluded that high [CO2] or [HCO3-] decreases the mitochondrial [NADH]/[NAD+] ratio in hepatocytes, resulting in a decreased [beta-OHB]/[AcAc] ratio.
Sex-related differences in meprin-A, a membrane-bound mouse kidney proteinase
Authors: Stroupe S. T. , Craig S. S. , Gorbea C. M. , Bond J. S.
Published: December 2017

To investigate the expression of meprin-A, a brush-border metalloproteinase in mouse tissues, immunohistochemical studies were conducted using a monoclonal antibody prepared against a purified form of kidney meprin-A form male mice. Kidney slices from female mice displayed markedly less immunoreactivity compared with similar preparations from male mice using this antibody. However, the specific activities of meprin-A in kidney homogenates and purified preparations of meprin-A from male and female mice were not significantly different. Western blots of kidney membrane proteins from several mouse strains indicated that the female form of meprin-A had a decreased mobility relative to the male form when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis; this difference could be eliminated by treatment of preparations with endoglycosidase F, which removes some asparagine-linked oligosaccharides. These data and lectin blots of membrane proteins indicate that there are differences in the glycosylation (specifically in the complex type oligosaccharides) of meprin-A in adult (8 wk old) male and female mice. Juvenile (3 wk old) male and female mice displayed similar amounts of immunohistochemical staining in kidney slices, as well as similar meprin-A electrophoretic mobilities and lectin affinities. Administration of 17 beta-estradiol to gonadectomized adult mice decreased the immunoreactivity of meprin-A in kidney slices and the electrophoretic mobility of meprin-A. These studies indicate that estrogens affect posttranslational modifications of meprin-A.
Insulin responsiveness of CK-M and CK-B mRNA in the diabetic rat heart
Authors: Popovich B. K. , Sayen M. R. , Dillmann W. H.
Published: December 2017

Decreased cardiac performance is a known complication of diabetes mellitus, but the detailed molecular mechanisms that are responsible for this contractile abnormality are only incompletely explored, and cardiac gene products of known function, which are markedly and actively insulin responsive, have not been described. Recently, we found that creatine kinase (CK) enzyme activity and CK-M subunit mRNA levels are decreased in the heart of rats with experimental diabetes mellitus. These abnormalities could be restored to normal with chronic insulin administration. The CK-M and CK-B genes are expressed in the heart, and we wanted to determine whether diabetes also induces a change in CK-B mRNA levels. Quantitation of CK-M and CK-B mRNA levels on Northern blots with specific cDNA probes showed that, in diabetic hearts, CK-B mRNA levels represent only 19.8% of control levels and are more markedly depressed than CK-M mRNA levels, which are 46.5% of control values. Acute injection of insulin led to a significant 1.6-fold increase in CK-M mRNA and a 2.2-fold increase of CK-B mRNA 5 h after insulin injection. CK-M mRNA levels were restored to normal within 12 h, but 48 h were required to restore CK-B mRNA levels to normal values. After 1 mo of insulin therapy, CK-B mRNA levels had risen 9.7-fold, exceeding normal values by 90%, whereas CK-M mRNA levels were at the normal level as previously shown. CK enzyme activity showed only a small response to insulin administration 48 h postinjection. Diabetes leads therefore to a marked lowering of CK-M and CK-B mRNA levels in the rat heart.(ABSTRACT TRUNCATED AT 250 WORDS)
Glucose availability and sensitivity to anoxia of isolated rat peroneal nerve
Authors: Strupp M. , Jund R. , Schneider U. , Grafe P.
Published: December 2017

The contrast between resistance to ischemia and ischemic lesions in peripheral nerves of diabetic patients was explored by in vitro experiments. Isolated and desheathed rat peroneal nerves were incubated in the following solutions with different glucose availability: 1) 25 mM glucose, 2) 2.5 mM glucose, and 3) 2.5 mM glucose plus 10 mM 2-deoxy-D-glucose. Additionally, the buffering power of all of these solutions was modified. Compound nerve action potential (CNAP), extracellular pH, and extracellular potassium activity (aKe) were measured simultaneously before, during, and after a period of 30 min of anoxia. An increase in glucose availability led to a slower decline in CNAP and to a smaller rise in aKe during anoxia. This resistance to anoxia was accompanied by an enhanced extracellular acidosis. Postanoxic recovery of CNAP was always complete in 25 mM HCO3(-)-buffered solutions. In 5 mM HCO3- and in HCO3(-)-free solutions, however, nerves incubated in 25 mM glucose did not recover functionally after anoxia, whereas nerves bathed in solutions 2 or 3 showed a complete restitution of CNAP. We conclude that high glucose availability and low PO2 in the combination with decreased buffering power and/or inhibition of HCO3(-)-dependent pH regulation mechanisms may damage peripheral mammalian nerves due to a pronounced intracellular acidosis.
In vivo evidence for hepatic autoregulation during FFA-stimulated gluconeogenesis in normal humans
Authors: Clore J. N. , Glickman P. S. , Nestler J. E. , Blackard W. G.
Published: December 2017

To examine the effect of increased gluconeogenesis [by increasing free fatty acids (FFA)] on hepatic glucose output (HGO) and on the first substrate (glucose) cycle, a primed continuous infusion of [2-3H]- and [6-14C]glucose was infused to isotopic steady state in 12 normal male volunteers after an overnight fast. Blood samples for the determination of glucose specific activity were obtained before and after an infusion of saline (n = 6) or 10% Intralipid and heparin (90 mU.kg-1.min-1, n = 6). Plasma FFA (593.3 +/- 74.5 to 971.1 +/- 127.1 mumol/l, P = 0.007) and glycerol (68.0 +/- 5.9 vs. 222.4 +/- 32.0 mumol/l, P = 0.002) increased during the lipid infusion, and beta-hydroxybutyrate levels rose from 0.24 +/- 0.12 to 0.50 +/- 0.17 mmol/l (P = 0.01). No change in plasma glucose, insulin, or glucagon levels was observed during the study, and levels of the gluconeogenic substrates alanine and lactate were also unchanged. Baseline rates of glucose cycling (rate of appearance of [2-3H]glucose minus rate of appearance of [6-14C]glucose) were similar in the two groups [1.44 +/- 0.33 vs. 1.33 +/- 0.44 mumol.kg-1.min-1, not significant (NS)] and did not change during either saline or lipid infusion, respectively. However, Cori cycle activity (the conversion of [6-14C]- to [1-14C]glucose) increased significantly from 0.59 +/- 0.19 to 1.28 +/- 0.19 mumol.kg-1.min-1 (P = 0.002) after FFA and glycerol levels had been increased, in marked contrast to the saline control (0.51 +/- 0.18 to 0.39 +/- 0.18 mumol.kg-1.min-1, NS).(ABSTRACT TRUNCATED AT 250 WORDS)
Neuronal influence on hormone release from anglerfish islet cells
Authors: Milgram S. L. , McDonald J. K. , Noe B. D.
Published: December 2017

Pancreatic islets in anglerfish (AF) are macroscopic collections of nearly pure endocrine cells that are densely innervated. Immunohistochemical staining for neurotransmitter biosynthetic enzymes revealed noradrenergic and cholinergic innervation of AF islets. An in vitro preparation of perifused dispersed AF islet cells was developed to study nutrient and neural control of islet hormone secretion. Glucose stimulated insulin and somatostatin-14 (SS-14) secretion in a dose-dependent manner, and 16.7 mM glucose inhibited glucagon secretion. In 2 mM glucose, norepinephrine and isoproterenol stimulated glucagon and SS-14 release. Isoproterenol stimulated insulin secretion, and norepinephrine stimulated or inhibited insulin release, depending on the concentration. Clonidine potently inhibited glucose-stimulated insulin secretion but stimulated glucagon release. Methacholine, a muscarinic cholinergic agonist, stimulated insulin, glucagon, and SS-14 release. The control of AF hormone release by neurotransmitter agonists in vitro was similar to that in higher vertebrate species. Therefore we used this tissue preparation to study postsynaptic interactions between glucose and neurotransmitters in islets.
Basal and insulin-mediated carbohydrate metabolism in human muscle deficient in phosphofructokinase 1
Authors: Katz A. , Spencer M. K. , Lillioja S. , Yan Z. , Mott D. M. , Haller R. G. , Lewis S. F.
Published: December 2017

Biopsies were obtained from the quadriceps femoris muscle of two male patients deficient in phosphofructokinase (PFK) 1. In the basal state the patients had markedly higher contents of UDP-glucose (approximately 5-fold), hexose monophosphates (approximately 7- to 13-fold), inosine monophosphate (IMP) (approximately 15-fold), and fructose 2,6-bisphosphate (F-2,6-P2; approximately 6-fold) than controls. Fructose 1,6-bisphosphate was not detectable, and phosphocreatine was lower (33 and 54 mmol/kg dry wt) than in controls [72 +/- 4 (SD)]. Patients had normal fasting plasma glucose and insulin levels and basal glucose turnover rates and responded normally to a 75-g oral glucose challenge. Patients were also studied during euglycemic hyperinsulinemia (approximately 95 mg/dl; 40 and 400 mU.m-2.min-1). Whole body glucose disposal rates were normal during both insulin infusion rates. Biopsies taken after the 400 mU insulin infusion showed decreases in acetylcarnitine and citrate and increases in the fractional activity of glycogen synthase. It is suggested that the high basal levels of F-2,6-P2 are, at least partly, a consequence of the high levels of fructose 6-phosphate, which will stimulate flux through PFK-2 and inhibit fructose-2,6-bisphosphatase. The low phosphocreatine and high IMP contents indicate that carbohydrate availability is important for control of high-energy phosphate metabolism, even in the basal state. The insulin-mediated decreases in acetylcarnitine and citrate suggest an activation of the tricarboxylic acid cycle in skeletal muscle but an absence of the normal response to replenish these intermediates.
Isolated hypoisoleucinemia impairs whole body but not hepatic protein synthesis in humans
Authors: Lecavalier L. , De Feo P. , Haymond M. W.
Published: December 2017

It is not known whether an acute decrease in the plasma concentration of any essential amino acid, as occurs during insulin infusion, impairs protein synthesis. To test this hypothesis in humans, selective hypoisoleucinemia or hypothreoninemia was induced by insulin infusion while maintaining normal or elevated plasma concentrations of the other amino acids via their selective infusion. The effects on protein synthesis were assessed using leucine kinetics and fractional synthetic rates of the two hepatic proteins albumin and fibrinogen. Results were compared with those of a combined insulin and complete amino acid infusion. Hypoisoleucinemia increased leucine oxidation (P less than 0.03) and decreased nonoxidative leucine disposal (P less than 0.04) and net leucine balance (P less than 0.03), whereas hypothreoninemia had no effect on any of these parameters. Neither hypoisoleucinemia or hypothreoninemia altered albumin and fibrinogen fractional synthetic rates when compared with the control study. Because of the known relationships between intra- and extracellular amino acid concentrations, the hypoisoleucinemia was most likely associated with a decreased intracellular concentration of isoleucine; such would not be the case for hypothreoninemia. Thus acute limited availability of a single essential amino acid can adversely affect nonhepatic protein synthesis.
1,2-Diacylglycerol and ceramide levels in rat liver and skeletal muscle in vivo
Authors: Turinsky J. , Bayly B. P. , O'Sullivan D. M.
Published: December 2017

1,2-Diacylglycerol and ceramide levels were measured in liver and skeletal muscle of rats under the following four experimental conditions: 1) during rapid fetal growth in the second half of gestation and during postnatal aging, 2) during tissue anoxia lasting up to 10 min, 3) during fasting for up to 6 days, and 4) during stress induced by injection of endotoxin (lipopolysaccharide from Salmonella enteritidis). 1,2-Diacylglycerol and ceramide levels in fetal liver were 77 and 58% lower, respectively, than those of young rats after weaning. 1,2-Diacylglycerol and ceramide concentrations in liver of postnatal rats were not influenced by aging. Anoxia produced a 41–64% increase in liver 1,2-diacylglycerol levels and a 9–21% increase in liver ceramides. Fasting of adult rats for up to 6 days had no effect on 1,2-diacylglycerol and ceramide levels in liver. However, fasting increased skeletal muscle concentrations of 1,2-diacylglycerol and ceramides 88 and 44%, respectively. Injection of endotoxin had a biphasic effect on liver 1,2-diacylglycerol levels, causing a transient 52% increase at 1 h, followed by a 24–38% decrease below the control level at 17 h after endotoxin administration. Liver ceramide levels were increased 66, 52, and 27% at 1, 3, and 5 h after endotoxin, respectively, but did not differ from control at 17 h after injection. Endotoxin had no effect on muscle 1,2-diacylglycerol and ceramide concentrations at any interval.
Postnatal changes in plasma 1,25-dihydroxyvitamin D3 in sheep: role of altered clearance
Authors: Ross R. , Dorsey J.
Published: December 2017

The ontogeny of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] production and metabolic clearance rates (PR and MCR, respectively) was determined in chronically catheterized fetal (postconceptional age of 127 days; term = 145 days), neonatal (10 +/- 1 and again at 24 +/- 1 days postnatal), and adult (3 yr of age) sheep. Plasma concentrations of 1,25(OH)2D3 were more than twofold higher in fetuses than in adults and were increased by 78% during the 1st 24 days postnatally. In contrast, compared with values in fetuses, the MCR of 1,25(OH)2D3 was reduced by 40% in 10-day-old neonates and by 58% in 24-day neonates, although 24-day neonatal values were still 75% higher than adult values. In contrast, the PR of 1,25(OH)2D3 did not demonstrate an ontogenic change during development and remained at all times approximately ninefold the adult value. The finding, by regression analysis of an inverse relationship between the MCR and the plasma concentration of 1,25(OH)2D3, is interpreted to suggest that increased plasma concentrations are the result of a reduced MCR. We conclude that increases in plasma concentrations of 1,25(OH)2D3 in the perinatal period can occur by a mechanism that is independent of enhanced endogenous synthesis, namely a reduction in the MCR of the hormone.
Regional blood-brain barrier transport of ketone bodies in portacaval-shunted rats
Authors: Hawkins R. A. , Mans A. M.
Published: December 2017

The permeability of the blood-brain barrier to ketone bodies, substrates of the monocarboxylic acid carrier, was measured in individual brain structures of control and portacaval-shunted rats. The measurements were made 5-7 wk after the shunt or sham operation by quantitative autoradiography. Portacaval shunting caused the permeability to ketone bodies to decrease throughout the brain by approximately 70%. There was a striking change in the transport pattern in the cerebral cortex; deeper cortical layers were affected more than superficial layers. Ketone body consumption by brain is limited by the transport capacity of the monocarboxylic acid system. Therefore, in portacaval-shunted rats the very low activity of this system makes it unlikely that ketone bodies can make a substantial contribution during situations such as fasting. Likewise, other substrates of the monocarboxylic acid system, e.g., lactate and pyruvate, will have greatly restricted access to the brain after portacaval shunting. If the carrier is symmetrical, another consequence will be that exit of endogenously produced lactate will be retarded.
Glutamine nitrogen kinetics in insulin-dependent diabetic humans
Authors: Darmaun D. , Rongier M. , Koziet J. , Robert J. J.
Published: December 2017

To assess the effect of insulin deficiency on whole body glutamine kinetics, five young adults with type I (insulin-dependent) diabetes received 4-h primed continuous infusions of L-[1-13C]leucine and L-[2-15N]glutamine in the postabsorptive state after blood glucose had been clamped overnight at either a normoglycemic level (approximately 85 mg/dl) or a moderate hyperglycemic level (approximately 260 mg/dl) by means of an automated glucose control insulin infusion system. The hyperglycemic state was associated with a significant rise in leucine level [from 165 +/- 23 to 242 +/- 62 (SD) microM], appearance rate (from 125 +/- 11 to 142 +/- 17 mumol.kg-1.h-1), and oxidation (from 27 +/- 10 to 31 +/- 10 mumol.kg-1.h-1). In contrast, neither the plasma level nor the appearance rate of glutamine (333 +/- 51 vs. 318 +/- 58 mumol.kg-1.h-1) was affected. We conclude that insulin deficiency resulting in moderate hyperglycemia induces a 13% rise in whole body proteolysis and yet does not stimulate glutamine de novo synthesis, despite increased precursor availability.
Reduced phospholipid contents of brain synaptosomes in phosphate depletion
Authors: Smogorzewski M. , Islam A. , Koureta P. , Fadda G. Z. , Massry S. G.
Published: December 2017

The effects of 6 wk phosphate depletion (PD) and pair-weight feeding (PW) without and with treatment with verapamil (PD-V and PW-V, respectively) on phospholipid (PL) and cholesterol content and on resting levels of cytosolic calcium concentration ([Ca2+]i) of brain synaptosomes of rats were examined. PD was associated with significantly (P less than 0.01) lower synaptosomal content of total PL, phosphatidylinositol (PI), phosphatidylserine (PS), and phosphatidylethanolamine (PE) and with significant (P less than 0.01) elevation in [Ca2+]i. Verapamil treatment of PD rats prevented the rise in [Ca2+]i of brain synaptosomes and the fall in PI and caused significant (P less than 0.01) improvement in the synaptosomal content of PS and PE; values of PS and PE in PD-V rats, however, were still significantly (P less than 0.01) lower than those of PD rats. Verapamil treatment of PW rats did not affect synaptosomal content of the various PL or resting levels of [Ca2+]i. Cholesterol content of brain synaptosomes of the various groups was not significantly different, but cholesterol-to-total PL ratios were significantly (P less than 0.01) higher in PD and PD-V rats than in PW or PW-V animals. Results indicate that PD affects metabolism of total phospholipids, PI, PS, and PE of brain synaptosomes, and these derangements are due to reduced availability of phosphorus and to the rise in [Ca2+]i. The data are consistent with the proposition that PD-induced changes in PL render the synaptosomal membrane more permeable to calcium, an event that may lead to a rise in [Ca2+]i.
Altered expression of muscle glucose transporter GLUT-4 in diabetic fatty Zucker rats (ZDF/Drt-fa)
Authors: Friedman J. E. , de Vente J. E. , Peterson R. G. , Dohm G. L.
Published: December 2017

We examined GLUT-4 glucose transporter protein and mRNA in muscle tissue from a new rodent model of non-insulin-dependent diabetes mellitus (NIDDM), the male obese Zucker diabetic fatty (ZDF) rat [ZDF/Drt-fa(F10)]. We also determined whether prevention of hyperglycemia might affect GLUT-4 expression by feeding the intestinal alpha-glucosidase inhibitor acarbose (40 mg/100 g diet) in the diet of male ZDF rats for 19 wk, starting at least 1 wk before the onset of diabetes. Fasting glucose was four- to sixfold greater in diabetic ZDF rats (24.1 +/- 6.7 mM) compared with lean or obese nondiabetic rats. Fasting insulin in diabetic ZDF rats (0.5 +/- 0.1 ng/ml) was similar to lean rats (0.4 +/- 0.1) but greatly reduced compared with obese nondiabetic rats (18.7 +/- 4.0 ng/ml). Acarbose treatment significantly reduced fasting glucose levels to 13.4 +/- 1.4 mM, while insulin levels increased to 1.6 +/- 0.3 ng/ml. GLUT-4 protein levels in diabetic ZDF rats were reduced approximately 40% in red quadriceps and mixed gastrocnemius muscles but were unchanged in white quadriceps muscle. Acarbose treatment was associated with a twofold increase in GLUT-4 protein and mRNA in mixed gastrocnemius muscle. These data indicate that, in this obese model of NIDDM without hyperinsulinemia, there is reduced muscle GLUT-4 protein in red but not white muscle fiber types. The decrease in muscle GLUT-4 expression in this model of NIDDM can be prevented by acarbose treatment, which reduces hyperglycemia and increases beta-cell responsiveness.