bluuebunny/crossref_metadata_2025 huggingface dataset:
Fatty acid metabolism in human fetal placental vasculature
Authors: Tulenko T. N. , Rabinowitz J. L.
Published: December 2017

The utilization of exogenous acetate and palmitate for the synthesis of vascular wall lipids was studied in isolated blood vessels of the human placenta under high- and low-oxygen conditions. Arteries and veins were obtained from fresh placental material and incubated in Krebs-Henseleit buffer (pH 7.4 at 37 degrees C) containing (in mM) either [2-14C]acetate, 0.11; [1-14C]palmitate, 0.43; or [16-14C]palmitate, 0.43. At the end of the 180-min incubation, neutral lipids accounted for 40%, and phospholipids 60%, of the 14C recovered. Following incubation with either precursor, 14C label was recovered mainly in 16-carbon fatty acids (FA) (35%) and 18-carbon FA (45%); less than 10% was recovered in less than or equal to 14-carbon FA and greater than or equal to 20-carbon FA. Estimates of synthesis rate however indicated that the 18- and 20-carbon FA had the highest turnover rates. Schmidt degradation analysis demonstrated considerable labeling of alkyl carbons after incubation with either carboxyl- or alkyl-labeled precursors, indicating that FA degradation products were used for de novo FA synthesis. Collectively, these findings suggest the presence of both de novo and chain elongation pathways. No differences were observed between arteries and veins, and anoxia had little influence on FA metabolism. Analysis of ATP levels demonstrated elevated concentrations of ATP present, accounting for the apparent lack of anoxic inhibition of FA metabolism.
Disparate effect of clomiphene and tamoxifen on pituitary gonadotropin release in vitro
Authors: Adashi E. Y. , Hsueh A. J. , Bambino T. H. , Yen S. S.
Published: December 2017

The direct effects of clomiphene citrate (Clomid), tamoxifen, and estradiol (E2) on the gonadotropin-releasing hormone (GnRH)-stimulated release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were studied in cultured anterior pituitary cells obtained from adult ovariectomized rats. Treatment of pituitary cells with Clomid or enclomid (10(-8) M) in vitro for 2 days resulted in a marked sensitization of the gonadotroph to GnRH as reflected by a 6.5-fold decrease in the ED50 of GnRH in terms of LH release from 2.2 x 10(-9) M in untreated cells to 3.6 x 10(-10) M. Treatment with E2 or Clomid also increased the sensitivity of the gonadotroph to GnRH in terms of FSH release by 4.3- and 3.3-fold respectively. Tamoxifen, a related antiestrogen, comparable to Clomid in terms of its ability to compete with E2 for pituitary estrogen receptors, was without effect on the GnRH-stimulated LH release at a concentration of 10(-7) M. Furthermore, tamoxifen, unlike Clomid, caused an apparent but not statistically significant inhibition of the sensitizing effect of E2 on the GnRH-stimulated release of LH. Our findings suggest that Clomid and its Enclomid isomer, unlike tamoxifen, exert a direct estrogenic rather than an antiestrogenic effect on cultured pituitary cells by enhancing the GnRH-stimulated release of gonadotropin.
Adipose cell size-function relationships: insulin binding and degradation
Authors: Cushman S. W. , Noda D. , Salans L. B.
Published: December 2017

125I-insulin binding and degradation have been studied in isolated rat adipose cells of increasing size. Binding at 24 degrees C reaches a steady state by 40-60 min in the absence of significant degradation. At 37 degrees C, binding reaches only a transient maximum by 10-15 min because insulin is rapidly degraded. Cellular enlargement is associated with increasing steady-state binding per cell at 24 degrees C, and increasing maximum binding and degradation per cell at 37 degrees C, in spite of increasing plasma insulin concentrations in the rats from which cells were prepared. Detailed steady-state studies at 24 degrees C, however, fail to delineate the mechanism of these alterations. Although dissociation experiments are consistent with the presence of a small degree of negatively cooperative binding site interaction, its magnitude is unchanged with increasing cell size. Furthermore, binding levels per unit cellular surface at 24 degrees C, at least at those insulin concentrations eliciting a biological response, remain relatively constant. None of the alterations in insulin binding observed here can explain the enlarged adipose cell's markedly decreased metabolic response to insulin.
Maternal and fetal adrenocortical function in the diabetic rabbit
Authors: Guleff P. S. , Beck R. R.
Published: December 2017

To investigate whether maternal or fetal glucocorticoid secretion is altered during pregnancy complicated by diabetes mellitus, in vitro production of corticosterone (B) and cortisol (F) was measured on day 28 of gestation in rabbits made alloxan diabetic prior to pregnancy. In addition, plasma B and F were followed throughout the experimental period. Fetuses of diabetic mothers were hyperglycemic and hyperinsulinemic, and mortality was much higher (18%) than that of control animals (1.5%). No significant differences were found in maternal plasma B or F during gestation, whereas F was significantly lower in fetuses of diabetic mothers. In vitro production of both B and F by maternal and fetal adrenals was lower in the diabetic animals, and responses to ACTH or insulin added to the incubation were the same in both groups. These results suggest that the ability of the adrenal cortex to secrete glucocorticoids may be impaired both in the mother and fetus when maternal diabetes is present. This could have effects both on maturational events in the fetus that are accelerated by glucocorticoids and on the ability to respond to stress.
Feedback regulation of thyrotropin by thyroxine under physiological conditions
Authors: Connors J. M. , Hedge G. A.
Published: December 2017

We have studied the contribution of thyroxine (T4) itself to the feedback regulation of thyrotropin (TSH) secretion. Thyroidectomized rats received T4 replacement by one of two methods: 1) 5 or 10 micrograms T4 . 100 g-1 . day-1 in drinking water or 2) continuous subcutaneous infusion of 0.5, 1.0, or 2.0 micrograms T4 . 100 g-1 . day-1. Replacement of 5 micrograms T4 . 100 g-1 . day-1 resulted in elevated plasma T4 and TSH while triiodothyronine (T3) was decreased. Replacement of 10 micrograms T4 . 100 g-1 . day-1 resulted in greater elevation of plasma T4, a transient decrease in T3, and no change in plasma TSH. Continuous replacement of T4 resulted in a dose-dependent elevation of plasma T4, little plasma T3 generation, and inhibition of the postthyroidectomy rise in plasma TSH. The pituitary responsiveness to thyrotropin-releasing hormone (TRH) was increased in groups receiving 1.0 or 2.0 micrograms T4 . 100 g-1 . day-1. It was concluded that 1) the plasma T4 exerts a negative feedback on basal TSH secretion in addition to that due to plasma T3 and 2) small amounts of T4 replacement enhance the TSH response to exogenous TRH in short-term hypothyroid rats.
Effects of altered adipose tissue morphology on plasma insulin levels in the rat
Authors: Schneider B. S. , Faust I. M. , Hemmes R. , Hirsch J.
Published: December 2017

The usual covariates of adiposity--adipocyte size, total body fat, and food intake--were experimentally dissociated using three new models of altered adipose tissue morphology in the rat. It was thereby possible to test the hypothesis that plasma immunoreactive insulin level (IRI) is a function of mean adipocyte size. In two of the models, experimental and control rats differ substantially in total body fat but show no difference in mean adipocyte size. In these models, no difference in plasma IRI was found between experimental and control animals. In a third model, experimental and control rats differ in mean adipocyte size but not in total body fat or daily food intake. In this model, plasma IRI was found to differ between experimental and control rats. These observations demonstrate a close link between adipocyte size and plasma IRI and suggest that the hyperinsulinemia and peripheral insulin resistance of obesity are more likely due to adipocyte hypertrophy than to increases in total body fat or daily food intake.
Induction of hepatic metallothionein by feeding zinc to rats of depleted zinc status
Authors: McCormick C. C. , Menard M. P. , Cousins R. J.
Published: December 2017

Induction of hepatic metallothionein was investigated in zinc-depleted rats after a single feeding (via stomach tube) of a complete diet with or without supplemental zinc. Subsequent to a maximum elevation in serum zinc (3.25 micrograms/ml) at 3 h, the rate of metallothionein synthesis increased 4.5-fold by 10 h after feeding. Changes in the rate of metallothionein synthesis coincided with similar changes in the level of translatable mRNA coding for metallothionein. Accumulation in liver of Zn2+ as metallothionein rose to a maximum by 12 h after diet administration and thereafter remained constant. Radioactive zinc (65Zn) included in the diet rapidly associated with newly formed metallothionein. Unlike Zn2+, 65Zn exhibited marked fluctuations within the 24-h period following feeding, indicating that zinc associated with metallothionein may be capable of exchange and/or dissociation. Changes in total liver 65Zn were accompanied by concomitant changes in metallothionein-bound 65Zn. This study has related (temporally) the metabolism of dietary zinc to the induction, apparently via transcription of mRNA, of hepatic metallothionein.
Intragastric pair feeding fails to prevent VMH obesity or hyperinsulinemia
Authors: Cox J. E. , Powley T. L.
Published: December 2017

Female rats with electrolytic lesions of the ventromedial hypothalamus (VMH) were pair fed with controls in a new paradigm in which food was delivered in normal temporal patterns, i.e., those of nonlesioned rats bar pressing for food. Lesions were produced after 6 days of adaptation to this regimen, and the experiment continued for 30 days postlesion. When they were killed, VMH rats contained substantially more carcass lipid than controls (20.4 vs. 12.4%) and were hyperinsulinemic (67.2 vs. 27.6 microunits/ml). Because these elevations developed in the absence of even minor disturbances in feeding patterns, these results support the view that VMH damage confers an essential metabolic bias favoring obesity. Separate VMH groups pair fed exclusively by either oral or intragastric routes exhibited virtually identical exaggerations in fat accumulation (18.4 and 17.7 g above controls, respectively) and plasma insulin (69.2 and 65.4 microunits/ml, respectively). Thus, contrary to a prediction of one form of Powley's (Powley, T. L. Psychol. Rev. 84:89–126, 1977) cephalic phase hypothesis, oropharyngeal stimulation and, hence, the cephalic responses of digestion are not critical for the metabolic disturbances that follow VMH damage.
Prior vagotomy blocks VMH obesity in pair-fed rats
Authors: Cox J. E. , Powley T. L.
Published: December 2017

Previously vagotomized, ventromedial hypothalamus (VMH)-lesioned rats and sham-lesioned controls were maintained on an intragastric pair-feeding regimen in which nonvagotomized VMH rats deposit excessive fat. Hypothalamic lesions were produced after 6 days of adaptation to pair feeding, and the experiment continued for 30 days postlesion. Extent of vagotomy was determined with a multiple-regression procedure with cell loss in the dorsal motor nucleus of the vagus, fasting gastric contents, and basal pancreatic protein output as predictor variables. The correlation was 0.95 between this set of indexes and the adequacy of a vagotomy for preventing hypothalamic obesity. Thus, radical vagotomies precluded the typical accumulation of significantly increased levels of carcass fat in lesioned animals (16.3 vs. 14.0% for controls). VMH rats with less extensive transections accumulated substantially more fat (25.9%). This outcome suggests that vagotomy produces a specific blockade of lesion-produced disturbances in metabolism leading to obesity. It fails to support a previous suggestion that vagal section blocks VMH obesity merely as a nonspecific surgical restriction of food intake because vagotomy was effective even though its effects on food intake could not operate.
Prostaglandins and renin release in vitro
Authors: Lin C. S. , Iwao H. , Puttkammer S. , Michelakis A. M.
Published: December 2017

The present studies were undertaken to explore further the role of prostaglandins in the release of renin from the renal cortex. To provide the best assessment of renin release, renin was determined by a radioimmunoassay for the direct measurement of renin. Slices of mouse renal cortex were incubated at 37 degrees C with arachidonic acid (AA), 5,8,11,14-eicosatetraenoic acid (ETA), indomethacin, prostaglandins, and synthetic prostaglandin endoperoxide analogue (EPA). Our results showed that AA at 1.5 X 10(-8) M significantly increased renin release at 10 and 30 min of incubation. This renin increase ws abolished by either ETA or indomethacin. Prostaglandin F2 alpha (PGF2 alpha) also significantly stimulated renin release at 10 and 60 min. PGE2 and 16,16-dimethyl PGE2 (DMPGE2) showed much less renin release-stimulating activity. EPA and PGI2 on the other hand very strongly stimulated renin release. However, at higher concentrations the stimulating effect of PGI2 and EPA disappeared and even became inhibitory in the case of EPA. Other prostaglandins were found to have no effect on renin release. The results suggest that the prostaglandin system directly affects renin release from the juxtaglomerular cells independent of systemic neurogenic and hemodynamic influences.
Porcine granulosa cells in suspension culture. II. Luteinization and hCG responsiveness
Authors: LaBarbera A. R. , Ryan R. J.
Published: December 2017

Granulosa cells from small follicles were cultured as suspensions in spinner flasks for 10 days in the absence or presence of follicle-stimulating hormone (FSH). With or without FSH, the cultured cells ultrastructurally resembled luteinized cells to different degrees. FSH increased progesterone accumulation in the culture medium. Ovine prolactin potentiated the effect of FSH in terms of the quantity of progesterone produced and the duration of accumulation. FSH increased acute human chorionic gonadotropin (hCG)-responsive progesterone secretion in short-term incubations of cultured granulosa cells. Responsiveness of FSH-cultured cells was maximal at day 4; that of control cultured cells was maximal at day 6. Adenylate cyclase activity of homogenates of cells cultured for 4, 6, or 8 days was measured. FSH induced in cultured cells an hCG sensitivity of the adenylate cyclase enzyme. These results indicate that FSH induced hCG-responsive progesterone secretion and hCG-responsive adenylate cyclase activity that correlate with ultrastructural signs of luteinization and with the previously reported FSH induction of hCG receptors.
Dose-response characteristics for effects of insulin on production and utilization of glucose in man
Authors: Rizza R. A. , Mandarino L. J. , Gerich J. E.
Published: December 2017

To determine the dose-response characteristics for the effects of insulin on glucose production, glucose utilization, and overall glucose metabolism in normal man, 15 healthy subjects were infused with insulin for 8 h at sequential rates ranging from 0.2 to 5.0 mU.kg-1.min-1; each rate was used for 2 h. Glucose production and utilization were measured isotopically ([3-3H]glucose). Tissue insulin receptor occupancy was estimated from erythrocyte insulin binding. Glucose production was completely suppressed at plasma insulin concentrations of approximately 60 microunits/ml. Maximal glucose utilization (10–11 mg.kg-1.min-1) occurred at insulin concentrations of 200–700 microunits/ml. The concentration of insulin causing half-maximal glucose utilization (55 + 7 microunits/ml) was significantly greater than that required for half-maximal suppression of glucose production (29 +/- 2 microunits/ml, P less than 0.01). Maximal effects of insulin on glucose production and utilization occurred at plasma insulin concentrations causing 11 and 49% insulin receptor occupancy, respectively. The above dose-response relationships indicate that in man 1) glucose production is more sensitive to changes in plasma insulin concentration than is glucose utilization; 2) both hepatic and peripheral tissues may contain “spare” insulin receptors; and 3) relatively minor changes in plasma insulin concentration or insulin receptor function can cause appreciable alterations in glucose metabolism.
Alloxan action on glucose metabolism in cultured fibroblasts. I. Stimulation and inhibition of glucose utilization
Authors: Ishibashi F. , Hidaka H. , Fields R. M. , Howard B. V. , Bennett P. H.
Published: December 2017

The influence of alloxan on mammalian cell glucose metabolism has been investigated using human diploid fibroblastic cells in culture. When cell monolayers were exposed to D-[14C]glucose, the presence of alloxan (0.31–1.87 mM) resulted initially in a dose-dependent enhancement of total cell glucose incorporation. This was observed within 2 min and declined by 6 min. After that time, alloxan inhibited glucose incorporation. When hexose transport was examined directly using glucose analogues, alloxan neither enhanced nor inhibited the uptake of 3-O-methylglucose or 2-deoxyglucose. Alloxan exerted no effect on cell permeability or cell viability. These results suggest that alloxan may directly influence cell glucose metabolism beyond the level of phosphorylation. The dual effect of alloxan on glucose incorporation may be related to the alloxan stimulation and subsequent inhibition of glucose-induced insulin release in pancreatic islets.
Simultaneous measurements of umbilical uptake, fetal utilization rate, and fetal turnover rate of glucose
Authors: Hay W. W. , Sparks J. W. , Quissell B. J. , Battaglia F. C. , Meschia G.
Published: December 2017

Fetal umbilical glucose uptake was compared with simultaneous measurements of glucose turnover and utilization rates in 12 pregnant sheep, at a mean of 137 days gestational age (range, 118–146 days). Umbilical glucose uptake was calculated by application of the Fick principle. Fetal glucose turnover rate was measured by a primed-constant infusion of [14C]- and [3H]glucose (glucose turnover rate = tracer infusion rate divided by fetal glucose sp act). The calculation of fetal glucose utilization rate required substraction of the loss of tracer to the placenta from the tracer infusion rate, thus defining the net tracer entry into the fetus for direct comparison with the net umbilical glucose uptake. In fed, normoglycemic sheep, these measurements demonstrated statistical equivalence of umbilical glucose uptake rate (4.77 mg.min-1.kg-1 +/- 0.34 SE) and glucose utilization rate ([14C]glucose, 5.58 mg.min-1.kg-1 +/- 0.54 SE; and [3H]glucose, 7.19 mg.min-1.kg-1 +/- 1.24 SE) when tested by two-way analysis of variance (P greater than 0.1). In three fasted, hypoglycemic sheep, the umbilical glucose uptake rate fell to 1.43 mg.min-1.kg-1 +/- 0.56 SE, which was considerably lower than the simultaneous glucose utilization rate ([14C]glucose, 4.78 mg.min-1.kg-1 +/- 0.48 SE; and [3H]glucose, 6.81 mg.min-1.kg-1 +/- 2.19 SE). Thus, in the normoglycemic, late-gestation fetal lamb, there appears to be little glucogenesis, whereas glucogenesis may become significant during fasting-induced fetal hypoglycemia.
Effects of hypophysectomy and thyroidectomy on leucine metabolism in adipose tissue
Authors: Coiro V. , Frick G. P. , Braverman L. E. , Goodman H. M.
Published: December 2017

Hypophysectomy doubled the rate of oxidation of L-[1–14C]leucine to 14CO2 by segments of rat epididymal adipose tissue. Thyroidectomy, but not adrenalectomy, produced identical results. Acceleration of leucine oxidation occurred even in the presence of glucose and saturating concentrations of insulin and leucine, suggesting that thyroidectomy increased the capacity to degrade leucine. Treatment of thyroidectomized rats with triiodothyronine (T3) decreased leucine oxidation, but at least 4 days were required. Treatment of hypophysectomized rats with T3 for 6 days was ineffective unless growth hormone was also given. A similar acceleration was also seen in the rate of oxidation of alpha-keto[1–14C]isocaproate, the deaminated analogue of leucine, but neither hypophysectomy nor thyroidectomy accelerated the rate of oxidation of isovalerate, the next metabolite in the degradative sequence. These observations suggested that hypothyroidism, whether primary or secondary, might increase the activity of the mitochondrial reaction responsible for the decarboxylation of alpha-ketoisocaproate. Because thyroidectomy failed to modify the rate of oxidation of [1–14C]pyruvate that occurs by an analogue reaction and requires the same cofactors, an effect of thyroidectomy on cofactor availability was ruled out. Direct assay in a cell-free homogenate revealed a nearly twofold increase in the activity of the alpha-ketoisocaproate dehydrogenase enzyme complex. The findings support the conclusion that hypothyroidism increases the amount or activity of the mitochondrial enzyme complex responsible for decarboxylation of branched-chain alpha-keto acids.
Inhibition of renin secretion by intrarenal alpha-adrenoceptor blockade
Authors: Blair M. L.
Published: December 2017

This study was designed to determine whether renal alpha-adrenoceptors can mediate tonic neural stimulation of renin secretion. The effect of alpha-adrenoceptor blockade by phenoxybenzamine (POB) or prazosin on renin secretion rate (RSR) was studied in pentobarbital-anesthetized dogs in which renal perfusion pressure was held constant with an adjustable aortic clamp. POB alone (100 micrograms.kg-1.min-1 iv) did not change arterial plasma renin activity (PRA). However, when beta-adrenoceptors were blocked by intravenous propranolol, intravenous POB infusion (100 micrograms.kg-1.min-1) decreased PRA and RSR to 48 +/- 8 and 21 +/- 9% of previous levels within 30 min. This effect was abolished by acute bilateral renal denervation. Direct intrarenal POB infusion (10 or 3.3 micrograms.kg-1.min-1) decreased RSR, whereas intravenous POB (3.3 micrograms.kg-1.min-1) had no effect on either RSR or PRA in propranolol-pretreated dogs. Prazosin (1 microgram.kg-1.min-1 iv) also significantly decreased PRA. These data indicate that when beta-adrenoceptors are blocked by propranolol, tonic neural stimulation of renin secretion is mediated by renal alpha-adrenoceptors.
Role of calcium in acute stimulated release of prolactin from neoplastic GH3 cells
Authors: Moriarty C. M. , Leuschen M. P.
Published: December 2017

Using a calcium-sensitive electrode to monitor calcium movements, we found that neoplastic GH3 cells experienced a net accumulation of calcium when exposed to elevated (50 mM) K+. Acute prolactin (PRL) release was also stimulated under these conditions. Both calcium uptake and PRL release could be blocked by the calcium antagonist methoxyverapamil (D-600). Thyrotropin-releasing hormone (TRH) also stimulated PRL release but had no effect on cellular calcium accumulation. Likewise, D-600 had no effect on TRH-induced PRL release. Such results indicate that enhanced secretory activity does not require an increase in intracellular calcium content. The observation that secretagogues do not stimulate PRL release in the absence of extracellular calcium was investigated. When GH3 cells were placed in a Ca-free medium, they underwent a prompt and sustained loss of cellular calcium. The loss of such intracellular calcium could be blocked with D-600. We conclude that the inability of TRH to stimulate the release of PRL in Ca-free medium is due to the loss of intracellular calcium and not to the absence of external calcium per se.
Photoperiodic regulation of growth: a photosensitive phase during light-dark cycle
Authors: Schanbacher B. D. , Crouse J. D.
Published: December 2017

Growth and performance of 72 young ram lambs were evaluated for susceptibility to exposure to three different photoperiods. Lambs were penned in groups of two and exposed to either short (8L:16D), long (16L:8D), or split (7L:9D:1L:7D) photoperiods. Lambs exposed to 16L:8D and 7L:9D:1L:7D photoperiods ate more feed (P less than 0.01), gained weight more rapidly (P less than 0.01), and required less feed per unit gain (P less than 0.01) than did lambs exposed to the 8L:16D photoperiod. Final weight and carcass weight were increased (P less than 0.01) as a result of exposure to 16L:8D and 7L:9D:1L:7D photoperiods; however, carcass attributes were not affected. Lambs exposed to the 7L:9D:1L:7D photoperiod had elevated prolactin concentrations that generally paralleled the prolactin profiles of lambs exposed to the 16L:8D photoperiod; however, concentrations of luteinizing hormone and testosterone were not affected by photoperiod. Lambs exhibited a daily feed-intake rhythm that was closely associated with daily photoperiod. These rhythms could account in part for the increased feed intake and performance of lambs exposed to 16L:8D and 7L:9D:1L:7D photoperiods. From these results, we conclude that 1) light is an important Zeitgeber for the regulation of feed intake, prolactin secretion, and growth rate in young lambs, and 2) a photosensitive phase exists for these functions during the light-dark cycle in sheep.
Nucleus ambiguus stimulation increases plasma insulin levels in the rat
Authors: Bereiter D. A. , Berthoud H. R. , Brunsmann M. , Jeanrenaud B.
Published: December 2017

The ventral lateral brainstem has been explored for sites that facilitate insulin release unilaterally, using electrical stimulation techniques in male rats anesthetized with alpha-chloralose/urethane. Stimulation in the region of nucleus ambiguus (amb) produced a rapid rise (by 1 min) in plasma insulin levels, whereas stimulation of brainstem regions further than 500 micrometers from amb had no consistent effect on insulin levels. The amb-induced rise in insulin was markedly exaggerated by the alpha-adrenergic antagonist, phentolamine, and was greatly diminished by bilateral cervical vagotomy or atropine pretreatment. These results strongly suggest that the amb is one source of vagal motoneurons that facilitate insulin secretion. However, amb electrical stimulation alone also activated an apparent sympathetic efferent output whose inhibitory effect on insulin release could be blocked by phentolamine.
Differential effects of cyclic nucleotide analogues and GnRH on LH synthesis and release
Authors: Liu T. C. , Jackson G. L.
Published: December 2017

We compared the effects of gonadotropin-releasing hormone (GnRH) and cyclic nucleotides on biosynthesis and release of luteinizing hormone (LH) by rat anterior pituitary glands (APs). APs from ovariectomized rats were incubated in the presence of test compounds, [3H]glucosamine, and [14C]alanine. GnRH significantly (P less than 0.01) increased both incorporation of [3H]glucosamine into LH (glycosylation) and release of immunoreactive LH (IR-LH) and [3H]LH. None of the cyclic nucleotides alone at 5 mM significantly stimulated release of IR-LH. Adenosine 3',5'-cyclic monophosphate (cAMP), N6,O2'-dibutyryl cAMP (DBcAMP), and 8-bromo-adenosine 3',5'-cyclic monophosphate (8-Br-cAMP) stimulated (P less than 0.01) glycosylation, whereas 8-Br-AMP, 8-bromoguanosine 5'-monophosphate (8-Br-GMP), cGMP, and 8-Br-cGMP had no detectable effect. Release of [3H]LH was increased by GnRH, 8-Br-cAMP, and DBcAMP, but not by other nucleotides. None of the cyclic nucleotides elevated [14C]LH, whereas GnRH elevated it (P less than 0.01) in the medium. Dose-response curves for glycosylation in response to 8-Br-cAMP and GnRH were parallel. In contrast, dose-response curves for release of both [3H]LH and IR-LH were nonparallel. It appears that cAMP can mimic the action of GnRH on LH glycosylation, but may have little direct effect on LH release.
Whole-body protein metabolism due to trauma in man as estimated by L-[15N]alanine
Authors: Birkhahn R. H. , Long C. L. , Fitkin D. , Jeevanandam M. , Blakemore W. S.
Published: December 2017

Changes in whole-body protein metabolism due to several skeletal trauma were estimated in five male volunteers and four male trauma patients using a single pulse of [15N]alanine and [13C]urea. The isotopic decay data were computer curve-fitted to obtain the best estimates for eight parameters of a four-pool model. The trauma patients excreted almost 5 times as much urinary nitrogen and utilized 33% more energy than did the volunteers on the same hypocaloric, protein-free diet. Trauma was found to expand the "active metabolic nitrogen" pool and the transfer rates of nitrogen into and out of a "slow turnover protein" pool. Whole-body protein synthesis increased 37% above normal and protein breakdown increased 79% above normal due to trauma. These values were in agreement with those determined at the same time using a [14C]leucine continuous infusion method. The data show that trauma accelerates the nitrogen flow and both protein synthesis and breakdown. This mechanism for negative nitrogen balance is consistent with that found for other severe stresses but is different from milder stresses.
Effects of fasting on flux and interconversion of leucine and alpha-ketoisocaproate in vivo
Authors: Nissen S. , Haymond M. W.
Published: December 2017

To determine directly the interconversion of circulating leucine and alpha-ketoisocaproate (KIC) in vivo, as well as the effects of fasting on leucine and KIC metabolism, 14- and 96-h fasted dogs were studied during simultaneous infusion of L-[4,5-3H]leucine and [U-14C]KIC. The specific radioactivities of 3H- and 14C-labeled leucine and KIC were determined and transfer rates calculated using a two-pool reversible model. In both groups, approximately 32% of the total leucine carbon entering was converted to KIC, whereas 60% of circulating KIC is converted to leucine. Plasma [3H]KIC specific radioactivity was only half of the circulating [3H]leucine specific activity, suggesting entry of KIC from an unlabeled pool. Fasting decreased the leucine-KIC interconversion, entry of unlabeled KIC, and irreversible loss of KIC. These data demonstrate that interconversion of circulating leucine and KIC is extensive and that fasting decreases the conversion of leucine to KIC in the labeled and unlabeled KIC pools, suggesting that transamination is a major regulator of leucine metabolism in fasting dogs.
Adipose tissue lipoprotein lipase and glycerol release in fasted Zucker (fa/fa) rats
Authors: Gruen R. K. , Greenwood M. R.
Published: December 2017

Fat cell hypertrophy occurs in most forms of obesity. In the obese Zucker rat, fat cell hypertrophy and increased lipoprotein lipase activity (LPL) are the earliest known signs of obesity. We studied the regulation of fat cell size in the obese Zucker rat by measuring changes in fat cell LPL activity and lipolysis in response to an overnight fast in 6- and 14-wk-old lean and obese rats. At both ages, fed obese rats had significantly increased fat cell size, LPL activity, and basal glycerol release in three adipose tissue depots compared to fed lean rats. Obese rats decreased LPL activity in response to fasting, but levels always remained equal to or greater than those in fed lean rats. Obese rats also showed a reduced lipolytic response to fasting. Thus, the obese rat after an overnight fast could not produce a coordinated response to fasting similar to the lean rat, and its homeostatic adjustments to this mild stimulus favored preservation of its enlarged fat cell size.
Metabolism of tritium- and 14C-labeled alanine in rats
Authors: Golden S. , Chenoweth M. , Dunn A. , Okajima F. , Katz J.
Published: December 2017

[3-3H]- and [U-14C]alanine were administered to starved rats by bolus injection and by continuous infusion. The specific activities of alanine, glucose, and lactate in blood were followed. The tracer kinetics of alanine depended on the site of tracer administration and sampling. Tracer was either administered into the aorta and blood sampled from the vena cava (A-VC mode) or tracer was administered into the vena cava and arterial blood sampled (V-A mode) (Katz, J. F. Okajima, and A. Dunn. Biochem J. 194: 513-524, 1981). When tracer was infused in the A-VC mode the plateau specific activity of alanine was about half that obtained in the V-A mode. The parameters of alanine turnover were calculated from the specific activities obtained in the A-VC mode. The calculated apparent replacement rate averaged 1.9 mg.min-1.kg-1 for [U-14C]- and 3.9 mg.min-1.kg-1 for [3-3H]alanine, indicating a carbon recycling of about 50%. The apparent contribution of alanine carbon to that of glucose is 15%. The maximal activity in plasma water is attained at about 5 min after bolus injection of [3-3H]alanine and that of [14C]glucose in blood is attained about 10 min after the injection of [U-14C]alanine. Maximal specific activity of [3H]- and [14C]lactate is attained within about 1 min after injection. The apparent mean transit time and alanine mass were calculated from the areas of washout curves after the continuous infusion was terminated. The mean transit time for [3H]alanine was 10 min and apparent total body mass of alanine of the order of 40 mg/kg. The apparent means transit time for [U-14C]alanine ranged from 33 to 66 min corresponding to a mass of the order of 100 mg/kg of alanine or 40 mg/kg of alanine carbon.
Chondrosarcoma growth: influence of diabetes, caloric restriction, and insulin treatment
Authors: McCumbee W. D. , Lebovitz H. E.
Published: December 2017

Diabetes and malnutrition result in decreased somatomedin production and cartilage growth in rats. The growth and metabolism of the Swarm rat chondrosarcoma are dramatically affected by somatomedins. Data presented here show that streptozotocin diabetes and diet restriction inhibit in vivo chondrosarcoma growth. Tumors grown in diabetic rats were significantly smaller than tumors grown in diet-restricted rats showing the same changes in body weight. Insulin treatment increased the rate of tumor growth in diabetic rats. Tumors grown in rigidly controlled diabetic rats were as large as tumors grown in nondiabetic controls. Diet restriction and diabetes reduced the capacity of the serum of the rat to stimulate alpha-amino[14C]isobutyrate uptake and [3H]uridine incorporation into RNA in chondrosarcoma pieces grown in nondiabetic rats. This somatomedin activity of the serum was restored by treating diabetic rats with insulin. There was a significant correlation between the in vitro stimulatory effect of a particular serum pool on chondrosarcoma metabolism and in vivo chondrosarcoma growth in the animals from whom the serum was obtained. These studies demonstrate that the in vivo growth of malignant chondrocytes is similar to that of normal chondrocytes with respect to the role of nutrition and insulin.
Decreased turnover of soluble liver proteins in mice with alloxan-induced diabetes
Authors: Duncan W. E. , Bond J. S.
Published: December 2017

Mice with alloxan-induced diabetes were used as a model to assess whether the synthesis and/or degradation of soluble liver proteins in general is affected in vivo by the diabetic state. Protein turnover was measured 2-3 wk after diabetes was induced. Degradation of liver cytosol proteins was decreased in diabetic mice as measured by the loss of protein radiolabeled with [14C]bicarbonate. The incorporation of radiolabeled amino acids into protein was also decreased in diabetic mice. When [3H]leucine was administered as the precursor for protein synthesis, the radiospecific activity of leucine derived from leucyl-tRNA in livers was similar in control and diabetic mice. Thus, the rate of protein synthesis appears to be decreased. There was no indication that diabetes affected the turnover of long- or short-lived proteins differentially. The activities of several cellular proteinases were unaffected or slightly decreased in livers of diabetic mice. These data indicate that protein turnover is decreased in this chronic form of diabetes.
Nuclear progestin receptor-like component in human cervical tissue
Authors: Holt J. A. , Pinkert C. A. , Lorincz M. A.
Published: December 2017

To study progestin action in human glandular cervix, we extracted the nuclear pellet with 0.5 M KCl and immediately adsorbed a progestin-binding receptor-like component to hydroxylapatite. Binding of 2 nM [3H]R5020 at 4 degrees C for 18 h, in the presence or absence of 100-fold molar excess progesterone and other steroids, was used to measure the total extractable and adsorbed progestin-binding component. With this assay, the amounts of progestin-binding component in the nuclear-rich fraction in specimens from women in the follicular and luteal phases of the ovarian cycle were 930 +/- 143 and 995 +/- 148 fmol/g wet wt (+/- SE), respectively; total cytoplasmic progestin receptor was 2,989 +/- 812 and 1,861 +/- 188 fmol/g wet wt (+/- SE) in the follicular and luteal phases, respectively. The progestin-binding component extracted from the nuclear pellet has a sedimentation velocity coefficient in the 3.5 S region when measured in high salt on 5-20% linear sucrose density gradients that contain 10% glycerol. The progestin binding measured by the hydroxylapatite-exchange assay with [3H]R5020 and radio-insert progesterone is high affinity (Kd 2.6 X 10(-10) M) and heat labile and appears specific for progestins and progesterone target tissues.
Transport of glucose analogues in rat lung
Authors: Kerr J. S. , Fisher A. B. , Kleinzeller A.
Published: December 2017

Uptake of nonmetabolizable glucose analogues was investigated in isolated rat lungs ventilated with 5% CO2 in air and perfused with Krebs-Ringer bicarbonate medium. In some experiments, [5–3H]glucose, methyl(alpha-D-[U-14C]gluco)pyranoside (alpha-MG), 3-O-methyl-D-[U-14C]glucose (3-O-MG), or various inhibitors were added to the initial perfusate to determine glucose utilization by the rate of 3H2O production or to characterize the uptake of glucose analogues. [1,2-3H]polyethylene glycol (PEG) was used as an indicator of the extracellular water space and gave a mean value of 0.35 ml/g tissue; calculated mean intracellular H2O space was 0.48 ml/g tissue. Glucose utilization was 56.4 +/- 6.6 (mean +/- SE, n = 6) mumol . g dry wt-1 . h-1 and decreased by 61% with 3 mM phlorizin. After 1–2 h perfusion, intracellular alpha-MG concentration was 1.4–1.9 times the extracellular concentration. The mean tissue-to-medium ratio (T/M) for alpha-MG decreased by more than 30% in the presence of glucose (5.0 mM), phlorizin (0.5 mM), ouabain (0.5 mM), or the absence of external Na+. Intracellular 3-O-MG concentration did not exceed extracellular concentration during 2 h of perfusion and the mean T/M did not change with any of the inhibitors studied. The results indicate that the nonmetabolizable glucose analogue alpha-MG is accumulated against a concentration gradient by an active Na+-dependent transport process, whereas 3-O-MG is apparently taken up by a different mechanism.
Muscle proteolytic enzyme activities in diabetic rats
Authors: McElligott M. A. , Bird J. W.
Published: December 2017

Proteolytic enzyme activities were measured in skeletal muscle of Sprague-Dawley rats with streptozotocin-induced diabetes [tail vein injection of streptozotocin (100 mg/kg), under ether anesthesia]. Assay of rat muscle homogenates from diabetic rats revealed a significant increase in alkaline serine protease activity as compared to untreated control rats and diabetic rats given insulin. There were no significant changes in lysosomal cathepsin activities in diabetic muscle as compared to controls. Gel studies of myofibrils isolated from the three groups of rats, subjected to autolysis, revealed that the serine protease had copurified with the myofibrils. Treatment of rats with compound 48/80, which degranulates mast cells, abolished the alkaline protease activity. There was no serine protease activity associated with the myofibrils isolated from compound 48/80-treated rats. Results from this study indicate that serine proteases are not involved in muscle protein breakdown in diabetes and are of mast cell origin.
Amino acid release from the hindquarter and urea appearance in acute uremia
Authors: Mitch W. E.
Published: December 2017

Hepatic urea production is increased in acutely uremic rats, but it is not known whether this is related to release of nitrogen from nonhepatic tissues. Rats with acute uremia had lower arterial concentrations of alanine, glutamine, and alpha-amino nitrogen when compared to sham-operated rats and released significantly more alpha-amino nitrogen from the hindquarter in situ. Release of alpha-amino nitrogen, alanine, and glutamine from the perfused hindquarter of acutely uremic rats was greater than that of sham-operated rats. These changes in situ and in the perfused hindquarter were more pronounced in rats deprived of food and water compared to fed animals and were not due to depletion of intracellular amino acids. In addition to increased amino acid nitrogen release, there was a higher urea appearance rate (excretion plus accumulation) in starved, uremic rats compared to sham-operated controls (244.7 +/- 11.2 vs. 182.0 +/- 12.4 mg. 100 g-1 .48 h-1); urea appearance also was suppressed partially by feeding. Both peripheral release of amino acids and diet influence waste nitrogen production in acute uremia.
Endotoxin-induced hyperinsulinemia and hyperglucagonemia after experimental liver injury
Authors: Cornell R. P.
Published: December 2017

Basal portal and systemic venous hyperinsulinemia and hyperglucagonemia were present in fasted rats at 24 h after experimental liver injury by partial (67%) hepatectomy, carbon tetrachloride gavage, and intravenous D-galactosamine administration. Both enhanced pancreatic hormone secretion and depressed hepatic hormone extraction were likely responsible for the insulin and glucagon oversupply. Endogenous gut-derived endotoxin is proposed as the causative factor for the exaggerated hormonal response because intravenous exogenous endotoxin elicited an identical elevation of insulin and glucagon. Systemic endotoxemia at 24 h after liver injury was indicated by marked (78–100%) lethality in lead-sensitized rats and positive Limulus lysate gelation tests of plasma samples. Furthermore, antiendotoxin treatments, including endotoxin tolerance, polymyxin B, and gut sterilization, significantly reduced both lead-sensitized lethality and hyperinsulinemic and hyperglucagonemic responses at 24 h in most liver-injury groups. Portal versus systemic venous administration of endotoxin at a low dose implied that normal endotoxin phagocytosis by the liver suppressed the pancreatic endocrine response. A physiological negative-feedback control system involving gut-derived systemic endotoxemia after liver damage with insulin and glucagon hypersecretion by the pancreas for stimulation of hepatic regeneration is hypothesized.
Insulin resistance for glucose metabolism in disused soleus muscle of mice
Authors: Seider M. J. , Nicholson W. F. , Booth F. W.
Published: December 2017

Our hypothesis was that insulin resistance for carbohydrate metabolism develops after a single day of muscular disuse. The immobilization of the mouse hindlimb for 24 h was used to produce muscular disuse (group c). As food intake was voluntarily decreased during the immobilization, two additional groups were used: group A was untreated and ate ad libitum, whereas group B was anesthetized with group C and was fed amounts of food similar to those eaten by group C. Because groups B and C differed only by limb immobilization, group B was used as the reference group. When insulin was present in the incubation media, the rates of 2-deoxyglucose uptake and glycogen synthesis were always significantly decreased in soleus muscles from group C (anesthetized, ate less, immobilized) as compared to group B (anesthetized, pair-fed food that group C ate). Significant interaction between the factors of insulin and muscular disuse for the rates of 2-deoxyglucose uptake and glycogen synthesis support the concept that disuse of skeletal muscle attenuates insulin action. These observations are a direct demonstration of a decrease in insulin responsiveness in skeletal muscle as the result of a single day of muscular inactivity. Because plasma insulin concentrations were significantly lower in groups B and C than in the untreated group, it seems likely that the development of insulin resistance in disused skeletal muscle is independent of plasma insulin levels.
Circadian profiles of insulin receptors in insulin-dependent diabetics in usual and poor metabolic control
Authors: Pedersen O. , Hjollund E. , Lindskov H. O. , Beck-Nielsen H. , Jensen J.
Published: December 2017

We have characterized the 24-h changes of insulin receptors on erythrocytes from patients with insulin-dependent diabetes of long duration. These diabetics were studied both in usual and poor metabolic control. Moreover, we have examined daytime changes of insulin receptors on monocytes from newly discovered diabetics. In both erythrocyte and monocyte studies, diabetics were compared to healthy controls. At insulin tracer concentration, insulin receptor binding to erythrocytes from diabetics in usual control and normal volunteers underwent a statistically significant diurnal variation with high binding values in the early morning, low daytime values with a nadir in the late afternoon, and a peak around midnight. Even diabetics in poor metabolic control due to insulin deprivation had preserved a similar 24-h rhythm of erythrocyte insulin receptors. Insulin receptor binding to monocytes at insulin tracer concentration declined significantly during the day both in newly discovered diabetics and in healthy controls. The mechanisms responsible for the acute phase changes of insulin-receptor binding are unknown, but the receptor changes seem related to the fed state. Moreover, analysis of the temporal interrelationship between erythrocyte insulin binding and plasma insulin concentration in diabetics during the 24-h period suggests that in these patients insulin may be one of the factors determining the rapid insulin receptor regulation.
Rate of bone loss in postmenopausal and osteoporotic women
Authors: Aloia J. F. , Ross P. , Vaswani A. , Zanzi I. , Cohn S. H.
Published: December 2017

Regional and total bone mass were determined in three groups of women by photon absorptiometry of the distal radius [bone mineral content BMC)] and total body neutron activation analysis [total body calcium (TBCa)], respectively. There were three groups of patients: group A, osteoporotic women treated with a variety of pharmacologic agents; group B, osteoporotic women (controls) taking only calcium supplements; and group C, normal postmenopausal women. The mean TBCa and BMC were considerably higher in the postmenopausal women than in the osteoporotic women. The rate of change of bone mass in group C was -0.45%/yr and -0.9%/yr for the total skeleton and radius, respectively. Group B had no significant rate of loss, whereas group A demonstrated a significant increase in TBCa of 0.75%/yr with no change in the BMC of the radius. There were no significant between-subject correlations for the slopes (rates of change) of the two bone mineral measurements.