bluuebunny/crossref_metadata_2025 huggingface dataset:
Hepatic uptake of insulin in man.
Authors: Tranberg K G
Published: December 2017

The hepatic uptake of unlabeled insulin (5--50 mU/kg) was determined in 45 nondiabetic patients without hepatic disease. Thirty patients were studied at normoglycemia and 15 at moderate steady hyperglycemia, induced by intraportal infusion of 80 or 200 mg glucose per h and per kg. The first pass fractional hepatic uptake of insulin (FH) was estimated by comparing the results after a portal and a peripheral infusion of unlabeled insulin in each patient. It varied negatively with the amount of insulin given, decreasing about 50% after a tenfold increase in dose. FH was markedly smaller during steady hyperglycemia than at normoglycemia, regardless of insulin dose. For low doses (5--10 mU/kg), giving insulin concentrations within the physiological range, FH was 0.43 (5 mU/kg) and 0.40 (10 mU/kg) at normoglycemia and 0.16 (10 mU/kg) at hyperglycemia. FH did not vary with the fasting or the glucose-stimulated, endogenous insulin levels recorded before insulin infusion. It is suggested that glucose is more important than insulin in modulating the fractional hepatic uptake of insulin.
Vasoactive intestinal polypeptide in vagally mediated pancreatic secretion of fluid and HCO3.
Authors: Fahrenkrug J , Schaffalitzky de Muckadell O B , Holst J J , Jensen S L
Published: December 2017

The role of nerves that liberate vasoactive intestinal polypeptide (VIP) in the porcine pancrease as mediators of the atropine-resistant action of the vagus on flow and bicarbonate (HCO3) secretion was examined. Efferent electrical stimulation of the vagus in atropinized pigs produced a profuse flow of pancreatic juice with high HCO3 content concomitantly with a significant increase in pancreatic VIP output from 13 to 113 fmol/min. Intravenous administration of somatostatin (SRIF) during continuous electrical vagal stimulation caused a parallel suppression of the VIP release and the pancreatic fluid and HCO3 secretion to prestimulatory values. The SRIF-induced reduction in fluid and HCO3 secretion seemed to be mediated via an inhibition of the VIP release rather than through a direct effect on the exocrine cells, inasmuch as SRIF did not influence the VIP-provoked exocrine response from the in vitro isolated perfused porcine pancreas. The results support the view that VIP is transmitter in the vagally induced atropine-resistant water and HCO3 secretion from the porcine pancreas.
Intestinal capillary blood flow during metabolic hyperemia.
Authors: Shepherd A P
Published: December 2017

It has been postulated that local circulatory control mechanisms regulate the O2 flux to parenchymal cells by two vascular mechanisms: changes in blood flow that minimize capillary PO2 variations and changes in the density of the perfused capillary bed through which O2 extraction is regulated. To test this prediction, isolated loops of canine jejenum and ileum were perfused at either constant blood flow or constant pressure, and intraluminal glucose was used to increase metabolic rate. In the constant-flow series, glucose increased O2 extraction, O2 uptake, and rubidium extraction. Resistance fell when the metabolic rate was elevated. In the constant-pressure series, glucose increased blood flow, O2 extraction, O2 uptake, and capillary filtration coefficients. These results show that vascular resistance falls and that capillary density increases following an increase in oxygen demand. Thus, the glucose-stimulated gut loop seems to be a valid model of metabolic hyperemia, and its behavior would be difficult to reconcile with a purely myogenic theory of intestinal blood flow autoregulation.
Pathways of cycloleucine transport in killifish small intestine.
Authors: Miller D S , Kinter W B
Published: December 2017

By using blocking concentrations of competitors, i.e., concentrations of other neutral amino acids that cause maximal inhibition, cycloleucine transport into slices or everted sacs of killifish (fundulus heteroclitus) small intestine could be partitioned into three pathways. One is apparently not mediated, a second is inhibited by all neutral amino acids tested (component 1), and a third, is inhibited by alpha-aminocarboxylic acids (component 2), but not by beta-alanine or taurine. Both mediated pathways were Na dependent, and each yielded a linear double reciprocal plot of initial slice uptake vs. cycloleucine concentration. Apparent Kt and Vmax values for component 1 were 0.03 mM and 33 pmol/mg tissue per 3 min, respectively; corresponding values for component 2 were 0.12 mM and 28 pmol/mg tissue per 3 min. Additional experiments with an intestinal brush border membrane vesicle preparation indicate that these mediated components reflect true differences in carrier specificity rather than the differential effects of inhibitors on metabolism or on the Na gradient that drives cycloleucine transport.
Mechanisms involved in alpha-adrenergic phenomena: role of calcium ions in actions of catecholamines in liver and other tissues
Authors: Exton J. H.
Published: December 2017

Epinephrine and norepinephrine binding sites with the physiological characteristics of alpha-adrenergic receptors have been identified in the plasma membranes of liver and other cells. Interaction of catecholamines with these receptors causes a mobilization of calcium ions from mitochondria and perhaps other intracellular stores in liver cells. In other cells, there may also be influx of extracellular calcium ions. Evidence is presented in support of the hypothesis that the rise in cytosolic calcium ions resulting from these changes is responsible for many of the alpha-adrenergic actions of catecholamines. Possible mechanisms by which activation of alpha-adrenergic receptors causes changes in calcium and other aspects of cellular metabolism are discussed.
Biological activity of protein-bound calcium in serum
Authors: Wortsman J. , Traycoff R. B.
Published: December 2017

We have previously reported the existence of a subfraction of serum calcium that is tightly bound to normal human serum proteins. This tightly bound calcium fraction (TBC) is thought to be a calcium-albumin-fatty acid complex (ACP) because similar complexes can be prepared by the sequential addition of albumin to calcium in the presence of palmitic acid. These studies deal primarily with TBC from rat serum and the uptake of calcium by bone cells in palmitate-treated serum. It is reported that TBC does not exchange with ionized calcium and that calcium binds strongly to albumin in the presence of palmitate. The uptake of calcium in palmitate-treated serum is three times greater than the uptake of calcium in control serum in both in vitro and in vivo systems. These findings demonstrate that 1) a tightly bound albumin-calcium fraction is present in both human and rat sera; 2) calcium in TBC may be complexed to albumin via fatty acids; 3) TBC does not participate in the maintenance of the level of ionized calcium in serum and 4) circulating TBC or ACP complexes may be taken up by living cells.
Sodium balance after adrenal enucleation
Authors: Eisenstein B. , Bengele H. H. , Alexander E. A.
Published: December 2017

To better define the relationship between adrenal enucleation (AE), sodium retention and adrenal regeneration hypertension (ARH), we measured daily sodium balance in rats after bilateral adrenal enucleation with (AEN) and without uninephrectomy (AE), and before and after development of ARH, along with the appropriate control groups. We found that both AE and AEN groups retained sodium during 7 days after surgery; AE 1.16 +/- 0.24 and AEN 1.32 +/- 0.33 meq/100 g body wt. Sham-operated, uninephrectomized rats and AE rats treated with spironolactone were in zero sodium balance. In the AEN rats after the development of ARH (BP greater than 150 Torr), zero sodium balance was also noted. Only AEN rats developed hypertension while sham-operated, uninephrectomized rats and AE rats with both kidneys remained normotensive for 28 wk. In summary, we have demonstrated that for 1 wk after AE, rats are in positive sodium balance and this is reversed by spironolactone but not by uninephrectomy.
Effect of alcohol ingestion on bone and mineral metabolism in rats
Authors: Baran D. T. , Teitelbaum S. L. , Bergfeld M. A. , Parker G. , Cruvant E. M. , Avioli L. V.
Published: December 2017

Chronic ethanol administration to growing rats for 56 days resulted in circulating levels of 140 mg/dl, approximating concentrations that characterize alcoholic intoxication in man. This degree of alcohol ingestion, although without gross or histological effect on the liver or testicles, was attended by decreased trabecular bone volume despite a normal rate of skeletal mineralization as measured by time-spaced tetracycline labeling. Concomitant serum levels of calcium, phosphate, magnesium, creatinine, glutamic oxalacetic transaminase, alkaline phosphatase, testosterone, and 25-hydroxyvitamin D were normal. Although alcohol treatment was associated with a significant decrease in urinary calcium excretion, it had no effect on phosphate excretion nor on its renal tubular reabsorption. The data reveal that circulating levels of alcohol that do not result in hepatic or testicular injury are toxic for bone.
Influence of thyroid status on skeletal muscle LPL activity and TG uptake
Authors: Kaciuba-Uscilko H. , Dudley G. A. , Terjung R. L.
Published: December 2017

The influence of thyroid status on the clearance of chylomicron 14C-labeled triglycerides (14C-TG), the fractional uptake of 14C-TG, and activity of lipoprotein lipase (LPL) in the different skeletal muscle fiber types was evaluated in pentobarbital-anesthetized rats. The turnover of plasma TG was approximately fourfold greater in the hyperthyroid (HyperT) compared to either euthyroid (EUT) or hypothyroid (HypoT) animals. The uptake of 14C-TG was increased in slow-twitch red and fast-twitch red muscle sections of the HyperT group and normal in the HypoT group. The changes in LPL activity in these two fiber types were inversely related to thyroid influence, with decreases found in the HyperT group and increases found in the HypoT group. Thus, the uptake of 14C-TG in these high-oxidative fibers, relative to the LPL activity, varied directly with thyroid influence. As a result, the normal linear relationship, apparent for the different types of skeletal muscle, between LPL activity measured in vitro and TG uptake determined in situ was not maintained.
Effects of human serum on transport of testosterone and estradiol into rat brain
Authors: Pardridge W. M. , Mietus L. J. , Frumar A. M. , Davidson B. J. , Judd H. L.
Published: December 2017

The effect in vivo of the plasma proteins in human serum on the transport of [3H]testosterone (T), [3H]-dihydrotestosterone (DHT), and [3H]estradiol (E2) through the brain capillary wall, i.e., the blood-brain barrier, was studied in anesthetized rats using a tissue-sampling-single-injection technique, In the absence of plasma proteins, approximately 90% of plasma T, DHT, or E2 was transported into brain on a single pass after a bolus carotid injection of labeled hormone. Serum was obtained from 57 patients in seven different clinical conditions: pregnancy, oral contraceptive use, thin and obese postmenopausal, follicular phase female, hirsutism, and normal male; the level (mean +/- SD) of sex hormone-binding globulin (SHBG) varied from 17 +/- 5 nM (hirsutism) to 323 +/- 83 nM (pregnancy). When the carotid injection solution was made 67% serum, the amount of T, DHT, or E2 transported into brain was inhibited in proportion to the concentration of SHBG. Among the patient groups, an overall linear inverse correlation between the mean SHBG level and the mean extraction of unidirectional influx of testosterone (r = 0.99) and estradiol (r = 0.98) was observed. These studies indicate that a) the undirectional clearance by brain of both testosterone and estradiol is inversely related to the SHBG level and b) the fraction of hormone transported into brain greatly exceeds the free (dialyzable) moiety and is essentially equal to the albumin-bound fraction of plasma testosterone or estradiol.
Insulin secretory dynamics during development of rat pancreas
Authors: Rhoten W. B.
Published: December 2017

The dynamics of insulin secretion during development of the fetal rat pancreas were investigated. The time of onset of glucose-induced insulin secretion was of special interest. Pancreases from 15- to 22-day-old fetal rats were perifused in vitro with low (0.5 or 0.9 mg/ml) or high (5 mg/ml) concentrations of glucose in the presence or absence of arginine and leucine. Levels of insulin in the perifusate were determined by radioimmunoassay. At day 17, a significant increase in perfusate insulin level was observed in response to arginine and leucine (each at 5 mM), This response was independent of a high concentration of glucose. In addition, perifusate insulin levels were augmented when the concentration of amino acids were kept constant and the glucose concentration was changed from a high level to a low level. On day 20, a monophasic, rapid-onset short-duration rise in insulin release with a high glucose concentration was observed. This response was enhanced by acetylcholine (2.7 x 10(-9) M). At days 21 and 22, insulin levels rose rapidly in the presence of high glucose and remained elevated. The results show that there is considerable precision in the timing of the onset and maturation of the glucose-induced insulin secretory response prenatally and reaffirm that insulin secretion by the fetal beta-cell varies with the stimulus applied.
Selective inhibition by neuraminidase of insulin action on hexose metabolism of mouse adipocytes
Authors: Tsuda M. , Taketomi S. , Iwatsuka H.
Published: December 2017

Mouse adipocytes treated with neuraminidase showed a decreased response of glucose oxidation to insulin although insulin binding to the cells was normal. The decreased response was associated with the release of sialic acids from the cells by enzyme digestion. The hormone action on 2-deoxyglucose uptake was also decreased. However, the hormone action on glyceride-glycerol synthesis or lipogenesis from glucose was unaltered when enzyme-treated cells were incubated with higher glucose concentration (greater than or equal to 5 mM). However, at lower glucose concentrations (< 5 mM), in which glucose transport was a rate-limiting step, the hormone action was markedly decreased. When fructose was used as a substrate, the enzyme-treated cells showed an impaired response to insulin in fructose oxidation but not in glyceride-glycerol synthesis and lipogenesis from fructose. These results suggest that the postreceptor systems of insulin action on glyceride-glycerol synthesis and lipogenesis from hexose are different from those of the hormone action on hexose transport and oxidation. Furthermore, alteration in insulin-sensitive metabolic profiles may be caused, in part, by changes in glycoproteins and/or glycoplipids on the cell surface.
Responses of reproductive and nonreproductive tissues to 17 beta-estradiol during ovine puerperium
Authors: Rosenfeld C. R.
Published: December 2017

The responses of regional blood flows and cardiac output to the systemic infusion of 17 beta-estradiol (E2) (1 microgram/kg) were studied with radionuclide-labeled microspheres in 11 chronically instrumented ewes 1-12 days after lambing. Blood flow to the uterine myometrium, endometrium, and caruncles increased 818 +/- 212, 1,149 +/- 376, and 544 +/- 160% (mean +/- SE, P < 0.025), respectively, within 90 min after the infusion of E2. There was, however, in each of these tissues a progressive increase in the magnitude of the response to E2 stimulation as the puerperium progressed, not attaining changes expected in the prepregnant ewe until the 12th postpartum day. Significant vasodilation also occurred in the fallopian tubes, cervix, vagina, ovaries, mammary gland, skin, and thyroid gland. The magnitude of the blood flow response in the tubes, cervix, and vagina increased as the puerperium progressed. Cardiac output rose 17 +/- 10% (P < 0.05) after the infusion of E2. Although the ovine reproductive tissues are sensitive to estrogen-induced vasodilation throughout the puerperium, the magnitude of the responses progressively increase, suggesting that vascular reactivity in these tissues is changing and is not similar to that of the prepregnant animal.
Age-dependent development of insulin resistance of soleus muscle in genetically obese (ob/ob) mice
Authors: Grundleger M. L. , Godbole V. Y. , Thenen S. W.
Published: December 2017

The relationship between insulin binding and its biological effects was studied in soleus muscle of 3- to 15-wk-old genetically obese (ob/ob) mice. At 3 and 4 wk of age, soleus muscle from lean and obese mice bound similar amounts of insulin under equilibrium binding conditions. However, by 6 wk of age, insulin binding and total receptor concentration (Ro) were significantly decreased in soleus muscle from obese compared to lean mice. In addition lean and obese mice demonstrated an age-dependent decrease in insulin binding, Ro, and receptor affinity. At 4 wk of age, insulin-stimulated 2-deoxyglucose transport and glucose utilization were significantly lower in soleus muscle from obese mice and preceded alterations in insulin binding. The postmembrane decrease in insulin sensitivity was dissimilar for various pathways of glucose metabolism. Glucose conversion of glycogen, but not the glycolytic rate or glucose oxidation, remained sensitive to insulin stimulation. These data indicate that glucose transport and utilization rather than insulin binding may play the primary role in the development of insulin resistance of muscle in the obese diabetic syndrome.
Free fatty acid utilization by skeletal muscle after endotoxin administration
Authors: Romanosky A. J. , Bagby G. J. , Bockman E. L. , Spitzer J. J.
Published: December 2017

The effect of endotoxin administration on free fatty acid (FFA) utilization by skeletal muscle was investigated. Albumin bound [1“14C]palmitate was continuously infused into anesthetized dogs. Blood samples from the carotid artery and profunda femoris veins were obtained and blood flow through the thigh muscles was determined. After endotoxin, skeletal muscle blood flow decreased by 29%, reflecting the average decrease in cardiac output. Arterial FFA concentration in endotoxin-treated animals was significantly reduced relative to saline-treated animals. Whole-animal FFA turnover decreased up to 37% after endotoxin. FFA uptake was linearly correlated with arterial FFA concentration both preceding and following endotoxin and the slopes of the regression lines were identical. FFA uptake by skeletal muscle was significantly reduced during the early postendotoxin time period. Skeletal muscle FFA oxidation after endotoxin also tended to decrease. It is concluded that the changes in skeletal muscle FFA metabolism are most likely due to decreased arterial FFA levels after endotoxin rather than a direct effect of endotoxin on skeletal muscle.
Corticosterone secretion after inhibition of prostaglandin synthesis by indomethacin
Authors: Thompson M. E. , Hedge G. A.
Published: December 2017

Systemic indomethacin (Ind) administration decreased prostaglandin F (PGF) content of the rat adrenal to less than 1.4 pg/mg. This was less than 5% of the adrenal PGF content in the gelatin-treated (Gel) control group (34 pg/mg). Basal plasma corticosterone levels were increased by the Ind treatment. Since the calculated metabolic clearance rate for corticosterone was unchanged, this increase was attributed to an enhanced adrenal secretion rate that was secondary to elevated plasma ACTH concentration. Ether exposure in the presence of Ind did not stimulate a normal rise in plasma corticosterone or adrenal corticosteroidogenesis. Adrenal responsiveness to exogenous ACTH was reduced after Ind treatment. There was a normal rise in plasma ACTH levels following ether exposure confirming the adrenal as the site of inhibition. Systemic Ind treatment thus appears to have two sites of action in altering plasma corticosterone levels: 1) a direct effect on the adrenal, inhibiting normal secretion in response to acute elevations of plasma ACTH, and 2) an action at the pituitary or hypothalamic level, eliciting an increase in basal ACTH secretion.
Differential effects of progesterone on secretion of gonadotropic hormones in the rhesus monkey
Authors: Resko J. A. , Ellinwood W. E. , Knobil E.
Published: December 2017

To obtain new information on the site of the inhibiting actions of progesterone (P) during the follicular phase of the cycle, we administered P continuously to 13 rhesus macaques (Macaca mulatta) from days 5 through 12 of the menstrual cycle. This treatment produced luteal-phase levels (approximately 6 ng/ml of serum) within 4 h. (FSH) concentrations dropped significantly from pretreatment amounts (P less than 0.01). During this time, both immunoreactive luteinizing hormone (LH) (n = 8 animals) and bioactive LH (n = 5) remained stable. The 17 beta-estradiol (E2) concentrations dropped significantly below base line 24 h after treatment (P less than 0.05). The decline in E2 occurred after the decline in FSH. Inhibition of FSH continued for 40 h, after which both FSH and LH rose steadily in a way that resembled a preovulatory surge on days 8 or 9 of the menstrual cycle, and then returned to base line by day 10. When gonadotropins were at their zenith, the ovary did not respond by secreting E2. None of the effects mentioned above were found in control animals treated with blank implants. These data demonstrate that P administered during the follicular phase inhibits FSH but not LH secretion. This initial inhibition probably occurs at a hypothalamic-pituitary site, but after 40 h direct inhibitory effects on the ovary cannot be ruled out. P stimulates the release of gonadotropin in female monkeys despite reduced production of E2 by the ovary.
Sensitivity of rat adenohypophyseal cells to estradiol and LHRH during long-term culture
Authors: Lagace L. , Labrie F. , Antakly T. , Pelletier G.
Published: December 2017

To determine possible effects of the time in culture on the responsiveness of the different pituitary cell types to estrogens, rat anterior pituitary cells were incubated up to 20 days in the presence or absence of 10 nM 17 beta-estradiol. Whereas spontaneous luteinizing hormone (LH) and thyroid-stimulating hormone (TSH) release decreased by 85-90%, follicle-stimulating hormone (FSH) and prolactin accumulation in medium were only 50% decreased after 20 days in culture, thus suggesting that the secretion of FSH and prolactin is less dependent on extrinsic stimulatory factors. Estradiol increased spontaneous LH release and its responsiveness to luteinizing hormone-releasing hormone (LH-RH) up to day 16 in culture, whereas the stimulatory effect of the estrogen on FSH secretion was significant only up to day 6. The stimulatory effect of estradiol on basal TSH release was seen up to day 8 in culture, whereas that on spontaneous prolactin release increased progressively after day 8 in culture up to the last time interval studied (20 days). As revealed by immunocytochemistry, the stimulatory effect of estradiol was not due to changes of cell growth.
Use of aromatic amino acids as monitors of protein turnover
Authors: Williams I. H. , Sugden P. H. , Morgan H. E.
Published: December 2017

Phenylalanine and tyrosine were metabolized by the perfused rat heart via a mitochondrial aminotransferase. When L-[alanyl-2,3-3H]phenylalanine and L-[alanyl-2,3-3H]tyrosine were used, release of 3H2O was progressive over 2 h of perfusion. Metabolism of L-[U-14C]phenylalanine to 14CO2 or production of 3H2O from L-[ring-2,6–3H]phenylalanine or L-[ring-2,6-3H]tyrosine was not detected. Although 3H2O production from L-[alanyl-2,3-3H]phenylalanine was rapid, net production of phenylpyruvate or other metabolites of phenylalanine was negligible. As a result, use of aromatic amino acids as monitors of protein turnover in heart muscle was validated. Production of 3H2O from L-[alanyl-2,3-3H]phenylalanine was catalyzed by a mitochondrial enzyme, which is thought to be aspartate aminotransferase (EC 2.6.1.1). The rate of 3H2O production by both intact and detergent-treated mitochondria exceeded that of phenylpyruvate by a factor of 10 and occurred in the absence of alpha-ketoglutarate. These data provide an explanation for the production of 3H2O from L-[alanyl-2,3-3H]phenylalanine by perfused rat heart without the concomitant production of [3H]phenylpyruvate.
A functional analysis of dopaminergic innervation of the neurohypophysis
Authors: Passo S. S. , Thornborough J. R. , Ferris C. F.
Published: December 2017

Dopaminergic neurons arising from cell bodies in the rostral arcuate nucleus of the hypothalamus have been shown to make axoaxonic contact with neurohypophyseal neurosecretory axons. In this study, electrical stimulation of the rostral arcuate nucleus depresses multiunit electrical activity recorded from neurosecretory axons within the neurohypophysis. After a single 5-s stimulus train, neurohypophyseal electrical activity is reduced to 6% of control. The superfusion of dopamine (5 micrograms/microliters) onto the neurohypophysis also has an inhibitory effect. Superfusion directly onto the neurohypophysis of the dopamine-receptor antagonist, pimozide (1 micrograms/microliters), abolishes the inhibitory effect of arcuate nucleus stimulation. These findings suggest that the dopaminergic innervation of the neurohypophysis may have an inhibitory influence on the release of neurohypophyseal hormones.
Effect of liposome-adipocyte interaction on hexose uptake and insulin action
Authors: Sandra A. , Fyler D. J.
Published: December 2017

The effect of unilammelar lipid vesicles composed of defined acyl chain phosphatidylcholines or binary vesicles comprised of dioleoyl phosphatidylcholine (DOPC) and a variety of other lipid species on insulin action in isolated rat adipocytes was studied. Cells were treated with vesicles and subsequently analyzed for basal and insulin-stimulated hexose uptake. Of the different vesicles tested, only those containing DOPC and phosphatidylserine (PS) markedly inhibited the effect of insulin. The inhibition was a function of PS concentration in the binary vesicle. Basal uptake, on the other hand, was stimulated by PS-containing vesicles. The changes in insulin-stimulated hexose uptake following interaction of adipocytes with DOPC-PS vesicles were accompanied by alterations in the Vmax of the uptake process and in affinity of insulin binding, although the similar ED50 values of the control and vesicle-treated groups suggest that the observed effects on insulin sensitivity may be mediated by an uncoupling of the insulin receptor from transport activation. Several lines of evidence are consistent with vesicle-cell fusion as the pathway of DOPC-PS uptake under these conditions. Fluorescence microscopic analysis of fat cells following their interaction with vesicles filled with fluorescent dye indicated that the internal contents of the adipocytes became generally labeled. Biochemical studies of cell fractionation after the interaction of adipocytes with radiolabeled vesicles and effects of metabolic inhibitors and cell fixatives on vesicle uptake also support the fusion pathway.
Starvation suppresses sympathoadrenal medullary response to cold exposure in rats
Authors: Avakian E. V. , Horvath S. M.
Published: December 2017

The effect of 2 days (55 h) of starvation on the functional state of the sympathetic nervous system (SNS) and adrenal medulla in male rats was evaluated in both normal (24 degrees C) and cold (4 degrees C) environments. Fasting (24 degrees C) significantly decreased NE turnover in heart and spleen, and the concentration of plasma epinephrine (E) and norepinephrine (NE). Cold exposure in the fed animals significantly increased NE turnover in the heart and plasma E, but had no effect on plasma NE or spleen NE turnover compared to normal (no stress) conditions. Cardiac NE turnover was 50% less in fasted cold-stressed animals than in fed cold-stressed animals. Plasma E remained at low levels. Plasma concentration of free fatty acids was significantly elevated in the fasted state in both warm and cold environments. These results suggest that 2 days of starvation in adult male rats suppresses the activity of the SNS and adrenal medulla and interferes with the normal adrenergic response to cold stress. Moreover, E appears not to be essential for mobilization of fat stores for energy metabolism in the fasted state in either warm or moderately cold environments.
Investigation of kinetics of integrated metabolic response to adrenergic blockade in conscious dogs
Authors: Wolfe R. R. , Durkot M. J. , Wolfe M. H.
Published: December 2017

We have used multiple isotope infusions to study the integrated response of glucose, fat, and protein metabolism to combined alpha + beta-adrenergic blockade in conscious, unstressed, fasting (15 h) dogs. The response to the blocking agents was evaluated both with and without control of the glucoregulatory hormones. The hormones were controlled at the basal level by infusions of somatostatin and metyrapone to block their secretion, and by the infusion of insulin, glucagon, growth hormone, and cortisol at physiological rates. We found that adrenergic blockade markedly inhibited lipolysis, as reflected by falls in glycerol and plasma FFA appearance. The decrease in fat mobilization after blockade resulted in a proportionate shift from fat as an energy substrate toward carbohydrate. Glucose production and oxidation were both enhanced after blockade. The source of the increased glucose production was presumably hepatic glycogen because urea production was presumably hepatic glycogen because urea production was unaffected and glycerol uptake was decreased. These results are consistent with the interpretation that basal adrenergic activity plays an important role in the mobilization of fat in fasting dogs. A secondary consequence of that action is apparently a diminution of glucose production and oxidation, although the mechanism responsible for the latter response is not clear.
Mineralocorticoid activity of 19-nor-DOC and 19-OH-DOC in toad bladder
Authors: Perrone R. D. , Schwartz J. H. , Bengele H. H. , Dale S. L. , Melby J. C. , Alexander E. A.
Published: December 2017

Adrenal enucleation is followed by a period of increased sodium reabsorption thought to be due to excess mineralocorticoid activity. However, it has not been demonstrated that increased production of any known mineralocorticoid accounts for this antinatriuresis. Recently, 19-hydroxydeoxycorticosterone (19-OH-DOC) was found in incubates of regenerating adrenal capsules 3-4 days postenucleation and 19-nordeoxycorticosterone (19-nor-DOC) was identified in the urine of rats with regenerating adrenals. Because it was possible that these hormones might play a role in the sodium retention after adrenal enucleation, we compared the mineralocorticoid activity of these steroids to aldosterone using the toad bladder. Using short-circuit current as a measure of sodium transport, we found that 19-OH-DOC (10(-8) M) had no significant effect on sodium transport. However, 19-nor-DOC (10(-8) M) increased sodium transport to a degree not different from aldosterone (10(-8) M). Furthermore, the onset of action, duration of activity, and inhibition of effect of 19-nor-DOC by spironolactone were not different from that of aldosterone. We conclude that 19-nor-DOC exhibits a significant effect on sodium transport and thus has the potential to play a role in the sodium retention following adrenal enucleation. Under the conditions of these studies, 19-OH-DOC exhibited no effect on sodium transport.
Ontogeny of sex differences in LH and FSH levels 48 h after castration in the rat
Authors: Lorenzen J. R. , Ramaley J. A.
Published: December 2017

Serum gonadotropin levels were measured 12, 24, and 48 h after gonadectomy in male and female rats (ages, 22--60 days) to assess when during development the rate of rise of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) after castration approximates that seen in the gonadectomized adult. In females serum LH levels 48 h after ovariectomy were increased above sham levels only when the ovaries were removed prior to vaginal opening. Ovariectomy on the day of vaginal opening or at older ages resulted in no increase in LH levels by 48 h after surgery. Serum FSH levels at 24 and 48 h after ovariectomy declined with increasing age at the time of ovariectomy. In males serum LH levels at 48 h after castration increased with increasing age at the time of gonadectomy. Serum FSH levels at either 12, 24, or 48 h after orchidectomy did not change appreciably with age at the time of surgery. It is concluded that the acute pituitary secretion of gonadotropins after removal of testes in the immature male resembles that seen in the mature male early in the course of the development of sexual maturity. In contrast, the acute pituitary secretion of gonadotropins after removal of the ovaries in the immature female does not resemble that seen in the ovariectomized adult until she is mature and capable of ovulating. Thus, the observed delay in the rise of LH seen in ovariectomized adults may be a function of some aspect of the hormonal changes associated with the estrous cycle.
Effects of amino acids and gastric inhibitory polypeptide on insulin release in dogs
Authors: Yovos J. G. , O'Dorisio T. M. , Pappas T. N. , Cataland S. , Thomas F. B. , Mekhjian H. , Carey L. C.
Published: December 2017

Insulin release following intravenous administration of an amino acid solution with and without a simultaneous infusion of varying amounts of porcine gastric inhibitory polypeptide (GIP) was studied in dogs. Group I received a 10-amino acid mixture (300 mosmol/kg iv) at 16.6 ml/min for 1 h; group II, amino acid mixture plus 0.5 micrograms.kg-1.h-1 porcine GIP; group III, amino acid mixture plus 1.0 micrograms.kg-1.h-1 of GIP; group IV (a and b) received either 0.5 or 1.0 micrograms.kg-1.h-1 of GIP alone. Compared to group I, groups II and III had a greater insulin response during the first 30 min of the infusion. Group] IV (a and b) showed no insulin release. Glucose concentrations showed no significant change in all groups. From these results, it is concluded that insulin release after intravenous infusion of an amino acid mixture plus GIP is greater than after amino acids or GIP alone. It appears that this effect is more pronounced in the early phase of insulin release.
Regulation of alpha-ketoisocaproate binding to albumin in vivo by free fatty acids
Authors: Nissen S. L. , Miles J. M. , Gerich J. E. , Haymond M. W.
Published: December 2017

The importance of alpha-keto acid binding to plasma proteins was investigated both in vitro and in vivo using alpha-ketoisocaproate (KIC), the alpha-keto acid of leucine. Gel chromatography indicated that 65% of the radioactivity comigrated with serum albumin. An ultrafiltration assay was developed to estimate the percentage of free and bound KIC. These percentages, along with total plasma KIC concentrations, were used to calculate the circulating concentrations of free and bound KIC. KIC or free fatty acids (FFA) displaced [14C]KIC bound to bovine albumin or whole plasma. KIC was totally displaced from plasma proteins by 10 mM oleate, stearate, and myristate; whereas the alpha-keto acids of isoleucine and value were 50 and 85%, respectively, as effective as KIC. To determine whether increased plasma FFA concentrations alter the binding of KIC to plasma proteins in vivo, five postabsorptive humans were infused with triglyceride and heparin during the simultaneous administration of somatostatin, glucagon, and insulin. During the FFA elevation, plasma leucine decreased by 9% (P less than 0.02). Total plasma KIC remained constant, whereas free KIC increased (P less than 0.02) and bound KIC decreased (P less than 0.001). These results indicate that KIC is bound to plasma albumin in vivo and suggests that FFA, by altering circulating free KIC concentrations, may influence protein metabolism in man.
Reduced norepinephrine turnover in brown adipose tissue of ob/ob mice
Authors: Knehans A. W. , Romsos D. R.
Published: December 2017

Obese (ob/ob) mice have a lower thermogenic capacity than lean mice. The possible role of brown adipose tissue (BAT) in this defect was investigated. Lean and obese mice were exposed to 33 (thermoneutral), 25, or 14 degrees C for up to 3 wk. BAT cytochrome oxidase activity and numbers of Na+-K+-ATPase enzyme units, enzymes involved in thermogenesis, were similar at 33 or 25 degrees C. Chronic exposure to 14 degrees C increased these enzymes 34 and 62%, respectively, in lean mice and nearly 150% in obese mice. Sympathetic nervous system activity, which stimulates thermogenesis in BAT, was evaluated by measuring norepinephrine (NE) turnover. At 25 degrees C, NE turnover rate in BAT of obese mice was only 40% as rapid as in BAT of lean mice. Chronic exposure to 33 degrees C depressed NE turnover in BAT of lean mice, but not in obese mice, whereas exposure to 14 degrees C accelerated NE turnover in both lean and obese mice. Lower sympathetic nervous system activity in BAT of obese mice at 25 degrees C is likely a major factor in their reduced nonshivering thermogenesis and resultant high efficiency of energy storage.
Energy balance of rats with lateral hypothalamic lesions
Authors: Corbett S. W. , Keesey R. E.
Published: December 2017

Rats with lateral hypothalamic (LH) lesions maintain body weight at a chronically reduced percentage of nonlesioned controls. An assessment of how they achieve energy balance at subnormal weight levels entailed a determination of both their energy intake and their energy expended or lost in processing ingested food, on basal heat production, on activity, and in feces or urine. It was found that the caloric intake and expenditure of LH-lesioned animals, though significantly lower than those of controls, were appropriate to the reduced metabolic body size (BW0.75) that they maintained. Likewise, energy expenditure in the LH-lesioned animals was normal in that the proportion of their ingested energy relegated to 1) basal metabolism, 2) the processing food, and 3) activity was the same as that of nonlesioned controls. Thus, unlike nonlesioned rats, which at lowered body weights both decrease their energy needs and reorder the pattern of energy expenditure, LH-lesioned animals display a normal pattern of energy utilization at reduced weight levels. These findings provide further evidence that lateral hypothalamic mechanisms play an important role in setting the level at which body weight is regulated.
Evidence for renin in rat brain: differentiation from other reninlike enzymes
Authors: Dzau V. J. , Brenner A. , Emmett N. L.
Published: December 2017

We observed that unfractionated rat brain extract incubated with substrate at pH 6.0 yielded 12 times the quantity of angiotensin I as incubations at pH 7.4, but the enzyme activity measured at pH 6 was not primarily due to renin. To examine the existence of renin in brain, we used three methods of affinity chromatography (pepstatin-, renin-specific antibody-, and alpha-casein-Sepharose) to fractionate the angiotensin I-generating enzymes in the brain. 1) Brain extract applied to renin-specific column eluted a peak of angiotensin-releasing activity (ARA) that had a pH optimum of 6.0. This ARA was inhibited by antirenin antibody. Another peak of ARA with a pH optimum of 4 appeared in the nonbound fraction. This peak was not affected by antirenin antibody and had acid protease activity. 2) Pepstatin affinity column elution with lithium bromide yielded an early ARA peak (pH optimum 6.5), inhibited by antirenin antibody and a later peak (pH optimum 4.0) not inhibited by antirenin antibody. The latter contained acid protease activity. 3) alpha-Casein-Sepharose column also separated neutral proteases and immunoreactive renin from acid protease capable of generating angiotensin. In summary, rat brain contains a host of angiotensin I-generating enzymes that can be detected and separated as neutral and acid proteases and immunoreactive renin depending on the pH of the assay and conditions of purification. These findings indicate the presence of an enzyme with immunoidentity to renin in rat brain but do not imply local biosynthesis.
Cardiac uptake and secretion of catecholamines during adrenergic stimulation in vivo
Authors: Halter J. B. , Kelley K. O. , Gould K. L.
Published: December 2017

ardiac uptake and secretion of epinephrine (Epi) and norepinephrine (NE) were studied at rest and during the response to systemic vasodilation with dipyridamole in conscious dogs. Chronically implanted catheters in the left atrium and anterior great coronary vein allowed simultaneous measurement of arterial and venous levels of NE and Epi across the segment of myocardium perfused by the left anterior descending coronary artery. Arterial Epi was greater than coronary venous Epi both at rest and during vasodilation. Arterial and venous NE levels were similar at rest. However, arterial NE rose more than venous NE during dipyridamole. Thus, there was net cardiac uptake of NE as well as Epi when systemic plasma catecholamine levels increased in response to vasodilation. Fractional extraction [(A-V)/A X 100)] of Epi by the heart was 50% and was not affected over a sevenfold range of coronary artery flow. Fractional extraction of NE (5-15%) was significantly lower than that of Epi, most likely due to cardiac secretion of NE. Cardiac NE secretion was estimated to increase fourfold during dipyridamole. Thus, cardiac tissue of trained dogs avidly takes up circulating catecholamines. Despite an apparent increase of NE secretion by the heart during the systemic adrenergic response to dipyridamole, the heart remained an organ of net NE uptake and therefore did not contribute to the observed increase of systemic NE levels
Selective inhibition of glucose-stimulated insulin release by dantrolene
Authors: Janjic D. , Wollheim C. B. , Sharp G. W.
Published: December 2017

Dantrolene sodium, which interferes with excitation-contraction coupling by inhibiting the Ca2+ release from sarcoplasmic reticulum in muscle, was used to investigate the role of stored calcium in the stimulation of insulin release by various secretagogues. Insulin release was measured simultaneously with 45Ca2+ uptake or 45Ca2+ efflux from isolated rat pancreatic islets. Glucose-stimulated insulin release was inhibited by dantrolene (10–100 microM) as was glyceraldehyde- or mannose-stimulated release. In contrast, dantrolene failed to inhibit insulin release stimulated by leucine, arginine, ouabain, potassium, or 3-isobutyl-1-methylxanthine. Although dantrolene lowered glucose-stimulated 45Ca2+ uptake, nonspecific blockade of voltage-dependent Ca2+ channels may not be a primary action of dantrolene because K+-stimulated 45Ca2+ uptake was not inhibited. Glucose utilization (3H2O formation) was unaffected by dantrolene, whereas glucose oxidation (14CO2 production) was decreased. In the absence of Ca2+, the glucose-inhibited 45Ca2+ efflux was unchanged. At normal Ca2+, dantrolene inhibited glucose-stimulated 45Ca2+ efflux and veratridine induced insulin release. This suggests an interference with mobilization of beta-cell calcium stores. The selective action of dantrolene on insulin release makes it an interesting tool for further studies on stimulus-secretion coupling.
Abnormal brown adipose composition and beta-adrenoceptor binding in obese Zucker rats
Authors: Levin B. E. , Comai K. , O'Brien R. A. , Sullivan A. C.
Published: December 2017

The composition, morphology, beta-adrenergic receptor binding, and in vitro lipolysis were examined in lean and obese, 5- to 6-mo-old male Zucker rat interscapular brown adipose tissue (IBAT). IBAT pads from obese rats were heavier (283%), had more lipid (700%), and more (75%)( and larger (83%) adipocytes than those from lean rats. Also, IBAT from obese rats had no multiloculated cells, and 50% of their IBAT adipocytes were the size of white fat cells. High affinity binding for (-)-[3H]dihydroalprenolol (KD, 15-18 nM), as well as the estimated KD values for binding and the 1/2 Vmax values for adrenergic agonist-induced lipolysis were similar in isolated IBAT cells from lean and obese rats. However, adipocytes from IBAT in obese rats had 75% fewer high affinity beta-adrenergic binding sites per cell (Bmax) compared to those in lean rats. These findings are most compatible with the infiltration of IBAT by white adipocytes. Such infiltration would be expected to reduce the overall thermogenic capacity of IBAT in obese Zucker rats and thereby contribute to the maintenance of their obesity.
Fetal fuels. V. Ketone bodies inhibit pyrimidine biosynthesis in fetal rat brain
Authors: Bhasin S. , Shambaugh G. E.
Published: December 2017

Ketonemic states complicating late pregnancy are accompanied by lower brain weights in the newborn. Potential mechanisms whereby ketone bodies might inhibit cell proliferation were therefore examined in the fetal rat brain slice by measuring their impact on the de novo pathway for pyrimidine biosynthesis. DL-beta-hydroxybutyrate (10.8 mM) and acetoacetate (5.4 mM) were both found to diminish the incorporation of NaH14CO3 into [14C]UMP by 30%. This effect was similar in fetal tissues from fed and 48-h starved mothers. Graded concentrations of DL-beta-hydroxybutyrate (1.4-43.2 mM) resulted in a progressive inhibition that could not be explained either by isotope dilution consequent to ketone body oxidation or by a generalized inhibition of protein synthesis. The inhibition was not reversed with 10 mM glutamine, the principal nitrogen substrate for de novo biosynthesis of pyrimidines. When the conversion of orotic acid into UMP was blocked with 6-azauridine, DL-beta-hydroxybutyrate (10.8 mM) inhibited the incorporation of NaH14CO3 into orotic acid by 28%. By contrast, maximally inhibitory concentrations of this ketone body (43.2 mM) had no effect on the incorporation of [6-14C]orotic acid into [14C]UMP. Is is concluded that ketone bodies inhibit the de novo biosynthesis of pyrimidines in fetal brain slices and that they do so at a site proximal to orotic acid formation.
Enhanced receptor-cyclase coupling and augmented catecholamine-stimulated lipolysis in exercising rats
Authors: Williams R. S. , Bishop T.
Published: December 2017

To test the hypothesis that alterations of adipocyte beta-adrenergic receptors provide a molecular mechanism for enhanced catecholamine-stimulated lipolysis in physically trained animals, we studied adipocytes derived from rats subjected to 14 wk of swimming and from sedentary controls. Peak glycerol release and peak adenylate cyclase activity in response to epinephrine were increased in swimmers to 255% (P less than 0.01) and 156% (P less than 0.01) of control values, respectively, but neither basal glycerol release, basal cyclase activity, NaF-stimulated cyclase activity, beta-receptor number, nor receptor affinity for [3H]dihydroalprenolol were altered. Epinephrine-stimulated adenylate cyclase activity remained increased in adipocytes from swimmers in the presence of theophylline or adenosine. In the absence of exogenous guanine nucleotide, we observed no differences in the dissociation constants for either the high-affinity (KD = 0.025 microM) or the low-affinity (KL = 11 microM) classes of binding sites for (-)-epinephrine, but the proportion of high-affinity sites was greater in membrane preparations from swimmers than from controls (74 vs. 42%; P less than 0.01). We conclude that receptor-cyclase coupling is enhanced in adipocytes from exercising rats, perhaps due to an improved ability of adrenergic agonists to form the guanine nucleotide reversible high-affinity agonist-receptor complex.