bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of intestinal factors on extraction of insulin in perfused rat liver
Authors: Ikeda T. , Yoshida T. , Honda M. , Ito Y. , Murakami I. , Mokuda O. , Tominaga M. , Mashiba H.
Published: December 2017

To study the direct effect of intestinal factors on hepatic extraction of insulin, an investigation was made into the extraction of insulin from isolated rat liver perfused with portal venous effluent (PVE) obtained from the isolated perfused rat intestine. Rat intestine was perfused with Krebs-Ringer bicarbonate medium for 45 min, and the PVE was collected from glucose-, lipid-, or NaCl-treated and untreated control intestines. The PVE, after adjustment of its glucose (180 mg/dl) and insulin (200 microU/ml) concentrations, was used as the perfusing medium for the liver of a different rat. The liver was perfused without recirculation with the PVE not containing insulin for 15 min and then perfused with the PVE containing insulin for the next 30 min. Insulin removal from liver perfused with PVE from lipid- or NaCl-treated intestine (52.6 +/- 5.4 or 46.6 +/- 4.1%) was similar to that from comparable controls (49.7 +/- 2.8 or 48.2 +/- 2.9%), respectively. However, that from glucose-treated intestine (39.7 +/- 6.2%) was significantly (P less than 0.01) lower than that from control intestine (51.1 +/- 2.5%). These results indicate that an intestinal factor secreted after glucose ingestion significantly reduces hepatic extraction of insulin and that at least a part of the incretin phenomenon is due to a decreased hepatic extraction of insulin after oral glucose administration.
Modulation of receptors and adenylate cyclase activity during sucrose feeding, food deprivation, and cold exposure
Authors: Scarpace P. J. , Baresi L. A. , Morley J. E.
Published: December 2017

Thermogenesis in brown adipose tissue (BAT) serves as a regulator of body temperature and weight maintenance. Thermogenesis can be stimulated by catecholamine activation of adenylate cyclase through the beta-adrenergic receptor. To investigate the effects of sucrose feeding, food deprivation, and cold exposure on the beta-adrenergic pathway, adenylate cyclase activity and beta-adrenergic receptors were assessed in rat BAT after 2 wk of sucrose feeding, 2 days of food deprivation, or 2 days of cold exposure. beta-Adrenergic receptors were identified in BAT using [125I]iodocyanopindolol. Binding sites had the characteristics of mixed beta 1- and beta 2-type adrenergic receptors at a ratio of 60/40. After sucrose feeding or cold exposure, there was the expected increase in BAT mitochondrial mass as measured by total cytochrome-c oxidase activity but a decrease in beta-adrenergic receptor density due to a loss of the beta 1-adrenergic subtype. This BAT beta-adrenergic receptor downregulation was tissue specific, since myocardial beta-adrenergic receptors were unchanged with either sucrose feeding or cold exposure. In contrast, food deprivation did not alter BAT beta-adrenergic receptor density. Forskolin-stimulated adenylate cyclase activity increased in BAT after sucrose feeding or cold exposure but not after food deprivation. The ratio of isoproterenol-stimulated to forskolin-stimulated adenylate cyclase activity decreased in the sucrose-fed and cold-exposed rats but not in the food-deprived rats. These data suggest that in BAT, sucrose feeding or cold exposure result in downregulation of beta-adrenergic receptors and that isoproterenol-stimulated adenylate cyclase activity was limited by receptor availability.
Involvement of vitamin D3 with cardiovascular function. II. Direct and indirect effects
Authors: Weishaar R. E. , Simpson R. U.
Published: December 2017

To determine whether the changes in cardiovascular function that accompany vitamin D3 deficiency are the direct result of hypovitaminosis D3 or a response to the hypocalcemia that accompanies vitamin D3 deficiency, rats were maintained for 9 wk on a vitamin D3-deficient diet containing either low (0.4%) calcium or high (2.5%) calcium to prevent hypocalcemia. Rats were also maintained on the low-calcium, vitamin D3-deficient diet for 9 wk and then transferred to diets designed to reverse hypocalcemia or vitamin D3 deficiency. The results demonstrate that the changes in in vitro cardiac contractile function that accompany vitamin D3 deficiency 1) could not be prevented by preventing the hypocalcemia that normally accompanies vitamin D3 depletion or 2) could not be reversed by restoration of serum calcium to normal levels after the initial period of vitamin D3 depletion. In contrast, the change in in vitro vascular muscle contractile function observed in vitamin D3-deficient, hypocalcemic rats could be prevented by maintaining serum calcium at normal levels and also reversed by restoration of serum calcium to normal after an initial period of vitamin D3 deficiency. These observations indicate that hypocalcemia does not account for the changes in cardiac contractile function that result from vitamin D3 depletion and suggest a direct role for vitamin D3 or its metabolite 1,25-dihydroxyvitamin D3 in regulating cardiac contractility. Possible mechanisms underlying this direct effect were also explored.
Evidence of a defect in insulin-receptor recycling in adipocytes from older rats
Authors: Trischitta V. , Reaven G. M.
Published: December 2017

Although insulin-stimulated glucose uptake is known to be decreased in adipocytes isolated from old obese rats, the cause of this defect is not totally understood. In the present study, we examined the possibility that insulin resistance is associated with defects in the intracellular processing of the insulin-receptor complex. Adipocytes were isolated from control (2-mo-old rats) and obese, insulin-resistant rats (12-mo-old rats), and the following measurements were made: 1) insulin-stimulated glucose uptake; 2) insulin binding; 3) insulin-receptor internalization and recycling; 4) accumulation of insulin within the cell; and 5) rate of loss of insulin from the cell. The results indicated that maximal insulin-stimulated glucose uptake was significantly reduced in adipocytes from obese, insulin-resistant rats (increase over basal value was 500 +/- 53% in obese rats and 1,200 +/- 96 in control rats, P less than 0.01). 125I-insulin (A14) binding (cell-associated radioactivity) and the internalization of the hormone-receptor complex were not different in the two groups of animals studied. In contrast, insulin-receptor recycling was significantly decreased in adipocytes from obese rats (72.0 +/- 6.1 vs. 93.6 +/- 2.6%, P less than 0.01). In addition, loss of intracellular radioactivity was significantly prolonged in insulin-resistant rats (t1/2 = 12.05 +/- 0.9 vs. 9.4 +/- 0.3 min, P less than 0.05). Thus adipocytes isolated from the older rats were resistant to the insulin effect on glucose uptake, and this defect was not associated with a reduction in insulin binding. However, there was a decrease in insulin receptor recycling, and this phenomenon may be related to the insulin resistance present in these cells.
Food restriction modulates beta-adrenergic-sensitive adenylate cyclase in rat liver during aging
Authors: Katz M. S.
Published: December 2017

Adenylate cyclase activities were studied in rat liver during post-maturational aging of male Fischer 344 rats fed ad libitum or restricted to 60% of the ad libitum intake. Catecholamine (10(-5) M epinephrine, isoproterenol, or norepinephrine)-stimulated adenylate cyclase activity increased by 200-300% between 6 and 24-27 mo of age in ad libitum-fed rats, whereas in food-restricted rats catecholamine response increased by only 58-84% between 6 and 30 mo. In ad libitum-fed rats, glucagon (10(-5) M)-stimulated enzyme activity also increased by 40% between 6 and 12 mo and in restricted rats a similar age-related increase was delayed until 18 mo. beta-Adrenergic receptor density (Bmax, determined from saturation binding of the beta-adrenergic antagonist l-[125I]iodopindolol) increased by 50% between 6 and 24 mo in livers from ad libitum-fed but not food-restricted rats and showed a highly significant correlation with maximal isoproterenol-stimulated adenylate cyclase activity over the post-maturational life span. Age-related increases in unstimulated (basal) adenylate cyclase activity and nonreceptor-mediated enzyme activation (by guanine nucleotides, fluoride, and to a lesser extent forskolin) were retarded by food restriction. The results demonstrate that food restriction diminishes a marked age-related increase in beta-adrenergic-sensitive adenylate cyclase activity of rat liver. Alterations of adrenergic-responsive adenylate cyclase with age and the modulatory effects of food restriction appear to be mediated by changes in both receptor and nonreceptor components of adenylate cyclase.
Effect of phorbol myristate acetate on secretion of parathyroid hormone
Authors: Morrissey J. J.
Published: December 2017

The influence of phorbol myristate acetate (PMA), an activator of protein kinase c, on the secretion of parathyroid hormone from collagenase-dispersed bovine parathyroid cells was tested. The cells were incubated at low (0.5 mM) or high (2.0 mM) concentrations of calcium in the medium, and the hormone secreted into the medium was measured by a radioimmunoassay that recognizes both intact and C-terminal fragments of hormone. At low calcium, the secretory rate averaged 32 +/- 3.8 ng.h-1.(10(5) cells)-1. The addition of 1.6 microM PMA did not affect secretion. At high calcium there was a significant suppression of secretion by 38% to 19.8 +/- 3 ng.h-1.(10(5) cells)-1. The addition of 1.6 microM PMA significantly stimulated hormone secretion to 35.8 +/- 8 ng.h-1.(10(5) cells)-1, a rate indistinguishable from low calcium. This stimulatory effect of PMA at high calcium was seen at PMA concentrations as low as 1.6 nM, did not occur with a biologically inactive 4 alpha-isomer of phorbol ester, and was independent of changes in cellular adenosine 3',5'-cyclic monophosphate levels. Examination of 32P-labeled phosphoproteins by two-dimensional gel electrophoresis revealed acidic proteins of approximately 20,000 and 100,000 Da that were phosphorylated at low and high calcium + 1.6 microM PMA but not at high calcium alone. The protein kinase c activity associated with the membrane fraction of parathyroid cells significantly decreased 40% when the cells were incubated at high vs. low calcium. The data suggest that calcium may regulate parathyroid hormone secretion through changes in protein kinase c activity of the membrane fraction of the cell and protein phosphorylation.
TeBG- and CBG-bound steroid hormones in rabbits are available for influx into uterus in vivo
Authors: Chaudhuri G. , Steingold K. A. , Pardridge W. M. , Judd H. L.
Published: December 2017

The metabolic clearance rate (MCR) of gonadal or adrenal steroid hormones in rabbits often does not bear the expected inverse relationship with hormone binding to testosterone-binding globulin (TeBG) or corticosteroid-binding globulin (CBG). This suggests TeBG or CBG may not impede steroid hormone delivery to tissues. The effects of rabbit plasma proteins on the influxes of 3H-labeled steroids from the circulation into the rabbit uterus were measured in vivo using a tissue sampling single-injection technique. In the absence of plasma proteins, estradiol (E2) and testosterone (T) were freely diffusible through the uterine microvasculature (i.e., extraction greater than 80%). The extractions of dihydrotestosterone (DHT) and corticosterone (B) ranged from 60 to 72%, while that of cortisol (F) was reduced at 40%. Rabbit serum exerted no inhibition of the influxes of the steroids tested. The influxes of T and B greatly exceeded the rates that would be expected if only the free and albumin-bound fractions estimated in vitro were diffusible in vivo. However, the extraction of [3H]corticosteroid-binding globulin or bovine [3H]albumin were low, consistent with little, if any, extravascular uptake of the plasma proteins. The results indicate both albumin-bound and globulin-bound steroid hormone are available for transport into the uterus in the rabbit in vivo without significant exodus of the plasma protein, per se.
Dissociating antifertility effects of GnRH-antagonist from its adverse effects on mating behavior in male rats
Authors: Bhasin S. , Fielder T. , Peacock N. , Sod-Moriah U. A. , Swerdloff R. S.
Published: December 2017

Antagonist analogues of gonadotropin-releasing hormone (GnRH-A) alone inhibit spermatogenesis in experimental animals, but concomitant decline in serum testosterone leads to abolition of mating behavior. We assessed if the antifertility effects of GnRH-A could be dissociated from its effects on mating behavior by combining it with a small dose of androgen. Seven groups of six adult male Wistar rats were treated for 70 days as follows: I) controls, II) GnRH-A alone (250 micrograms/day), III) GnRH-A + 0.05 mg testosterone enanthate (TE), IV) GnRH-A + 0.15 mg TE, V) GnRH-A + 0.50 mg TE, VI) GnRH-A + 1.50 mg TE, and VII) GnRH-A alone (recovery group). Testes, prostate, and seminal vesicle weights were markedly reduced by GnRH-A treatment alone. Doses of TE required to maintain prostate and seminal vesicle weights were between 0.15 and 0.50 mg. Testis weights were not restored to normal even by the highest dose of TE. Intratesticular sperm counts were markedly decreased by GnRH-A treatment and restored only at the highest dose of TE (1.50 mg). Five out of six animals in group II, six out of six animals in group III, and one out of six in group IV were azoospermic. When mated with normal females, all animals in groups I and VI were fertile, all animals in groups II, III, and IV were infertile, whereas only four out of six animals in group V were fertile. All measures of mating behavior were impaired by GnRH-A treatment and restored by the smallest dose of TE (0.05 mg).(ABSTRACT TRUNCATED AT 250 WORDS)
Effect of hypophysectomy on energy balance and brown fat activity in obese Zucker rats
Authors: Holt S. , Rothwell N. J. , Stock M. J. , York D. A.
Published: December 2017

Hypophysectomy (HYPX) in genetically obese (fa/fa) Zucker rats significantly reduced body weight and energy gains and stimulated energy expenditure (by 34%), the thermic response to food (by 144%), and brown adipose tissue (BAT) mitochondrial GDP-binding capacity (by 190%) compared with pair-fed, sham-operated obese rats. These changes in energy balance in obese HYPX rats were reversed by corticosterone replacement (1 mg/day), but the increased BAT activity was only partly restored to normal. HYPX had only small effects on energy balance in lean Zucker rats compared with pair-fed, sham-operated lean controls but increased the acute thermic response to food and BAT mitochondrial GDP-binding capacity; these effects were inhibited by replacement of HYPX rats with corticosterone. The results suggest that alterations in the hypothalamic-pituitary-adrenal axis play a fundamental role in the development and maintenance of genetic obesity.
Adaptive changes in individual acyl-CoA esters from hamster BAT during cold acclimation
Authors: Donatello S. , Spennetta T. , Strieleman P. , Woldegiorgis G. , Shrago E.
Published: December 2017

Long-chain fatty acyl-CoA esters (LCFACoAE) were extracted from freeze-clamped powdered brown adipose tissue (BAT) obtained from thermoneutral control and cold-acclimated hamsters and the CoA esters individually separated by high-performance liquid chromatography. LCFACoAE of carbon chain length C12 to C20 were identified by increasing column retention time in the following order: C12:0, C14:1, C14:0, C16:1, C18:2, C16:0, C18:1, C18:0, and C20:4. The mean total LCFACoAE concentrations were 235 +/- 40 nmol/g protein for the control hamsters and 648 +/- 105 nmol/g protein for the 22-day cold-acclimated hamsters. A rapid fourfold increase in the levels of C16:0, C18:0, and C18:1 occurred within hours after initiation of the cold temperature, whereas the concentrations of the other six LCFACoAE either increased only slightly or remained unchanged. Almost 50% of the total LCFACoAE in the BAT of cold-acclimated hamsters was made up of C16:0, C18:0, and C18:1. These results, which demonstrate some dynamic changes in adipose tissue LCFACoAE, are consistent with their proposed role in the initiation and maintenance of BAT thermogenesis.
Leucine incorporation into mixed skeletal muscle protein in humans
Authors: Nair K. S. , Halliday D. , Griggs R. C.
Published: December 2017

Fractional mixed skeletal muscle protein synthesis (FMPS) was estimated in 10 postabsorptive healthy men by determining the increment in the abundance of [13C]-leucine in quadriceps muscle protein during an intravenous infusion of L-[1-13C]leucine. FMPS in our subjects was 0.046 +/- 0.003%/h. Whole-body muscle protein synthesis (MPS) was calculated based on the estimation of muscle mass from creatinine excretion and compared with whole-body protein synthesis (WBPS) calculated from the nonoxidative portion of leucine flux. A significant correlation (r2 = 0.73, P less than 0.05) was found between MPS (44.7 +/- 3.4 mg.kg-1.h-1) and WBPS (167.8 +/- 8.5 mg.kg-1.h-1). The contribution of MPS to WBPS was 27 +/- 1%, which is comparable to the reports in other species. Morphometric analyses of adjacent muscle samples in eight subjects demonstrated that the biopsy specimens consisted of 86.5 +/- 2% muscular as opposed to other tissues. Because fiber type composition varies between biopsies, we examined the relationship between proportions of each fiber type and FMPS. Variation in the composition of biopsies and in fiber-type proportion did not affect the estimation of muscle protein synthesis rate. We conclude that stable isotope techniques using serial needle biopsies permit the direct measurement of FMPS in humans and that this estimation is correlated with an indirect estimation of WBPS.
Effect of physical exercise on sensitivity and responsiveness to insulin in humans
Authors: Mikines K. J. , Sonne B. , Farrell P. A. , Tronier B. , Galbo H.
Published: December 2017

The effect of acute physical exercise on insulin sensitivity and responsiveness of glucose uptake and hepatic glucose production was studied. Seven untrained men were subjected to four sequential euglycemic hyperinsulinemic clamps after rest (R), immediately after exercise (E), as well as 48 h after 60 min of 150 W ergometer exercise (ER). Insulin-mediated glucose uptake was higher on E and ER days compared with R days. Apparent Km decreased after exercise (52 +/- 3 R vs. 43 +/- 4 E and 40 +/- 3 ER microU/ml, means +/- SE) and Vmax increased (9.5 +/- 0.8 R vs. 10.9 +/- 0.7 E and 10.7 +/- 0.8 ER mg.min-1.kg-1). Glucose oxidation increased with the increasing insulin infusion rate, and maximal glucose oxidation rate was lower on E days compared with R days. The maximal conversion rate of glucose to glycogen was higher on E and ER days (8.0 +/- 0.3 and 7.2 +/- 0.2, respectively) than on R days (5.7 +/- 0.6 mg.min-1 kg-1). Muscle glycogen synthase I activity was higher immediately after exercise and remained higher for the next 48 h. No change in any glucoregulatory hormone or metabolite could explain the increased insulin action seen after exercise. In additional experiments (n = 3), no remaining effect existed 5 days after exercise. Both insulin and exercise suppressed the pancreatic secretion of insulin and proinsulin. The conclusions drawn are that prolonged moderate exercise increases insulin action on glucose uptake in humans by reducing apparent Km and increasing Vmax. This effect lasts 48 h but not 5 days. The increased insulin action may be related to an exercise-induced increase in glycogen synthase activity.
Free and total catecholamines in critical illness
Authors: Woolf P. D. , Hamill R. W. , Lee L. A. , McDonald J. V.
Published: December 2017

The magnitude of circulating free and total (free plus conjugated) catecholamine responses to a strong and sustained stimulus was investigated in four groups of patients: traumatic brain injury (n = 24), vascular brain injury (n = 10), polysystem trauma (n = 7) and medical/surgical patients in an intensive care unit (n = 29). Despite significant three- to sevenfold elevations in both free and total norepinephrine (NE) and epinephrine (E), the ratio of free to total NE and E remained constant over a very broad range of values. The proportion of free E was twice normal (30.1-33.5 vs. 16.2%) in all but patients with polytrauma, whereas the percentage of free NE was unchanged in all patients (43.0%). All dopamine (DA) parameters remained generally normal. In the patients with traumatic or vascular brain injury, significant inverse correlations were present between the degree of neurological dysfunction, as indicated by the concomitant Glasgow coma score, and free NE, E, and DA and total NE and DA levels. Thus, during conditions of intense and prolonged catecholamine release, the proportion of free catecholamine remains constant and the total as well as free catecholamine concentration is proportional to the Glasgow coma score.
Increased in vivo glucose utilization in 30-day-old obese Zucker rat: role of white adipose tissue
Authors: Krief S. , Bazin R. , Dupuy F. , Lavau M.
Published: December 2017

In vivo whole-body glucose utilization and uptake in multiple individual tissues were investigated in conscious 30-day-old Zucker rats, which when obese are hyperphagic, hyperinsulinemic, and normoglycemic. Whole-body glucose metabolism (assessed by [3-3H]glucose) was 40% higher in obese (fa/fa) than in lean (Fa/fa) rats, suggesting that obese rats were quite responsive to their hyperinsulinemia (140 vs. 55 microU/ml). In obese compared with lean rats, tissue glucose uptake (assessed by the 2-deoxyglucose technique) was increased by 15, 12, and 6 times in dorsal, inguinal, perigonadal white depots, respectively; multiplied by 2.5 in brown adipose tissue; increased by 50% in skin from inguinal region but not in that from cranial, thoracic, or dorsal area; and increased twofold in diaphragm but similar in heart, in proximal intestine, and in total muscular mass of limbs. Our data establish that in young obese rats the hypertrophied white adipose tissue was a major glucose-utilizing tissue whose capacity for glucose disposal compared with that of half the muscular mass. Adipose tissue could therefore play an important role in the homeostasis of glucose in obese rats in the face of their increased carbohydrate intake.
Metabolic fate of a gastric glucose load in unrestrained rats bearing a portal vein catheter
Authors: Smadja C. , Morin J. , Ferre P. , Girard J.
Published: December 2017

Metabolic and hormonal responses to the administration of a gastric load of glucose (2.4 mg/g body wt) were evaluated after a fast of 6-7 h in unrestrained rats bearing a portal vein catheter. For the purpose of this study, we have designed a new technique for a direct catheterization of the portal vein that allows serial blood sampling in unrestrained Wistar rats. On the 6th postoperative day, food intake, body weight gain, liver function, histology of the liver, pancreas, intestine, and splanchnic blood flows were similar to those of sham-operated control rats. In the basal state glucose turnover was 21.3 +/- 0.9 mg.kg-1.min-1. After glucose ingestion, a portoperipheral gradient of lactate was present, reflecting the production of lactate by the intestine. Insulin secretion was biphasic and peaked at 12 min (344 +/- 46 microU/ml), whereas portal glucagonemia (400-500 pg/ml) remained unchanged. Sixty minutes after gastric glucose administration, 50% of the load was delivered at the periphery, and glucose utilization was increased by 100%. Hepatic glucose production decreased after 20 min and was inhibited by 30% at 60 min. Liver glycogen concentration remained unchanged during the experiment despite a normal capacity for glycogen synthesis.
Role of Ca2+ in response of aldosterone to stimulation by angiotensin II in vivo
Authors: Johnson E. I. , McDougall J. G. , Coghlan J. P. , Denton D. A. , Scoggins B. A. , Wright R. D.
Published: December 2017

The role of Ca2+ in stimulation of aldosterone secretion (ASR) has been evaluated in vivo using conscious sheep with an adrenal cervical autotransplant. The calcium antagonists verapamil, nisoldipine, and lanthanum and the calcium ionophore BAY K 8644 were infused into the adrenal arterial supply before or concomitantly with angiotensin II. Nisoldipine reversed stimulation of ASR (n = 4) from 13.6 +/- 3.2 to 4.8 +/- 1.2 nmol/h (P less than 0.01; control 2.3 +/- 0.6 nmol/h), as did verapamil. Lanthanum had an intermediate effect. In contrast, pretreatment with nisoldipine (n = 5) did not affect the response to angiotensin II, with ASR being 3.8 +/- 0.9 nmol/h after nisoldipine alone and 12.8 +/- 1.3 nmol/h after nisoldipine plus angiotensin II. In response to graded infusion of angiotensin II, nisoldipine blunted (P less than 0.01) to a small degree the response at all doses of the peptide. Close adrenal arterial infusion of the ionophore BAY K 8644 similarly reversed stimulation of ASR by angiotensin II. It also blocked the initiation of response to the peptide. These data are consistent with the involvement of two pools of calcium in the zona glomerulosa response to angiotensin II, an intracellular pool that is primarily responsible for the initiation of response and a transmembrane extracellular pool that is primarily involved in the "sustained" response to angiotensin II.
Pituitary and gonadal responsiveness is enhanced during GnRH-induced puberty
Authors: Spratt D. I. , Crowley W. F.
Published: December 2017

We hypothesized that the hypothalamic gonadotropin-releasing hormone (GnRH) signal that initiates sexual maturation is further amplified at both the pituitary and gonadal levels during puberty. To test this theory, six GnRH-deficient men were monitored during administration of exogenous GnRH at a physiological frequency for greater than or equal to 9 mo. GnRH doses were progressively increased until normal testosterone (T) concentrations and secondary sexual development were achieved. This "optimized" dose of GnRH was then sustained for at least 6 mo to allow maturation of the hypothalamic-pituitary-gonadal axis. The GnRH dose was then progressively decreased to a level that had been unable to stimulate normal T secretion before sexual maturation. Changes in pituitary responsiveness were analyzed in four of the six men by comparing gonadotropin responses to identical doses of GnRH before and after sexual maturation. Mean serum luteinizing hormone and follicle-stimulating hormone levels as well as luteinizing hormone pulse amplitudes were greater after the induction of sexual maturation than before despite identical doses of GnRH. Both pituitary and gonadal responsiveness was then analyzed in the remaining two subjects by choosing periods of evaluation where endogenous gonadotropin levels were matched before and after the period of sexual maturation. Serum T concentrations were greater after sexual maturation than before despite equivalent gonadotropin input to the testes and LH pulse amplitudes. Thus the testicular responsiveness to gonadotropins increased during sexual maturation. After initiation of puberty by GnRH secretion, amplification at both the pituitary and gonadal levels contributes to sexual maturation in the human.
Albumin gene transcription is enhanced in liver of nephrotic rats
Authors: Yamauchi A. , Imai E. , Noguchi T. , Tanaka T. , Yamamoto S. , Mikami H. , Fukuhara Y. , Fujii M. , Orita Y. , Kamada T.
Published: December 2017

The level of albumin mRNA and the transcription rate of the albumin gene were studied in the liver of control rats and rats with nephrosis induced by injection of the aminonucleoside of puromycin. Total RNA was extracted from liver by the guanidium thiocyanate method. The albumin mRNA level was measured by cDNA-RNA dot-blot hybridization, and the transcription rate of the albumin gene was measured by the "run-on" transcription assay using isolated nuclei. Urinary protein excretion in nephrotic rats was significantly higher than in control rats (258 +/- 132 vs. 12 +/- 2 mg/day), and the serum albumin concentration in nephrotic rats was significantly lower. There was no difference in body weight, liver weight, serum creatinine, or urea nitrogen between the two groups. Both the level of albumin mRNA and the transcription rate of the albumin gene in the nephrotic liver were about twice as high as those in the control liver. There was no difference in the level of beta-actin mRNA between the two groups. Northern blot analysis showed that both putative precursor RNA and the mature form of albumin mRNA were increased in nephrotic rats. We conclude that albumin synthesis is increased in the liver of nephrotic rats and a transcriptional process is responsible for this increase.
In vitro renal eicosanoid production during pregnancy in rabbits
Authors: Brown G. P. , Venuto R. C.
Published: December 2017

The low renal resistance to blood flow in the presence of an activated endogenous renin-angiotensin system in gravid animals may in part be mediated by the action of eicosanoids produced in situ. To evaluate intrarenal eicosanoid production during gestation in rabbits, we quantitated immunoreactive PGE2, 6-keto-PGF1 alpha (a stable metabolite of PGI2) and thromboxane B2 (a stable metabolite of thromboxane A2) in unextracted media after incubation of renal slices and isolated glomeruli. In cortical slices from nonpregnant and pregnant rabbits, PGE2 production (micrograms.g-1.30 min-1) was 0.04 +/- 0.005 and 0.08 +/- 0.01 (P less than 0.01) and 6 keto-PGF1 alpha was 0.03 +/- 0.01 and 0.06 +/- 0.01 (P less than 0.05), respectively. In papillary slices, PGE2 production was 14 +/- 2 and 21 +/- 2 (P less than 0.05) and 6 keto-PGF1 alpha was 4 +/- 1 and 5 +/- 1 (P greater than 0.05) for nonpregnant and pregnant rabbits, respectively. Thromboxane B2 production was unchanged during pregnancy in both cortex and papilla. Acute captopril administration to nonpregnant and to pregnant rabbits in vivo failed to alter in vitro renal slice eicosanoid production. Isolated glomeruli from nonpregnant and pregnant rabbits synthesized PGE2 at similar rates. Exogenous arachidonic acid increased PGE2 production (P less than 0.05), but angiotensin II had no effect on eicosanoid production in vitro. These data suggest that the net synthesis of vasodilator eicosanoids is enhanced during gestation in rabbits.(ABSTRACT TRUNCATED AT 250 WORDS)
Fatty acid-independent inhibition of hepatic ketone body production by insulin in humans
Authors: Keller U. , Gerber P. P. , Stauffacher W.
Published: December 2017

To investigate whether elevated plasma insulin or glucagon concentrations are capable of modifying hepatic ketogenesis independently of plasma free fatty acid (FFA) concentrations, ketone body production was determined by [3–14C]acetoacetate infusions in overnight-fasted normal subjects during exogenous supply of FFA (Intralipid and heparin infusion). When plasma FFA concentrations were elevated from 0.73 +/- 0.07 to 1.53 +/- 0.16 mmol/l during low insulin concentrations (approximately equal to 13 microU/ml) in group A (n = 7), total ketone body production increased from 3.6 +/- 0.6 to 8.2 +/- 1.0 mumol.kg-1.min-1 (P less than 0.001). When plasma FFA were similarly elevated during raised plasma insulin concentrations (approximately equal to 110 microU/ml) in group B (n = 5), total ketone body production was only 3.8 +/- 0.8 mumol.kg-1.min-1 (P less than 0.01 vs. group A). Low plasma FFA and low insulin concentrations resulted in total ketone body production of 0.70 +/- 0.18 mumol.kg-1.min-1 in group C (n = 7; P less than 0.01 vs. groups A and B). Elevation of plasma glucagon during Intralipid infusion in group D (n = 7) failed to affect ketogenesis, but the beta-hydroxybutyrate-to-acetoacetate concentration ratio decreased significantly (P less than 0.01). The data indicate that elevation of plasma insulin to high physiological concentrations restrains FFA-induced ketogenesis.
Slow to fast alterations in skeletal muscle fibers caused by clenbuterol, a beta 2-receptor agonist
Authors: Zeman R. J. , Ludemann R. , Easton T. G. , Etlinger J. D.
Published: December 2017

Chronic treatment of rats with clenbuterol, a beta 2-receptor agonist (8–12 wk), caused hypertrophy of histochemically identified fast- but not slow-twitch fibers within the soleus, while the mean areas of both fiber types were increased in the extensor digitorum longus (EDL). In contrast, treatment with the beta 2-receptor antagonist, butoxamine, reduced fast-twitch fiber size in both muscles. In the solei and to a lesser extent in the EDLs, the ratio of the number of fast- to slow-twitch fibers was increased by clenbuterol, while the opposite was observed with butoxamine. The muscle fiber hypertrophy observed in the EDL was accompanied by parallel increases in maximal tetanic tension and muscle cross-sectional area, while in the solei, progressive increases in rates of force development and relaxation toward values typical of fast-twitch muscles were also observed. Our results suggest a role of beta 2-receptors in regulating muscle fiber type composition as well as growth.
Role of gastric inhibitory polypeptide in postprandial hyperinsulinemia of obesity
Authors: Roust L. R. , Stesin M. , Go V. L. , O'Brien P. C. , Rizza R. A. , Service F. J.
Published: December 2017

To assess the contribution of gastric inhibitory polypeptide (GIP) to the postprandial hyperinsulinemia of obesity, secretion rates of GIP (generated from kinetic analyses from infusions of porcine GIP) and insulin (from C-peptide applied to a validated kinetic model) to meals of 3 sizes were determined in 10 obese (5 male and 5 female) and 10 lean, sex- and age-matched healthy subjects. Although the postprandial secretion rates of GIP were greater in obese subjects (P = 0.03), postprandial concentrations of GIP were not. The latter may be explained by the greater volume of distribution of GIP in obese subjects (P = 0.036). Secretion rates and volume of distribution of GIP were correlated (r = 0.652, P less than 0.01). Despite excessive integrated postprandial (P = 0.010) insulin concentrations, insulin secretion was not significantly different between obese and lean subjects. We conclude that 1) although postprandial plasma GIP concentrations are normal, GIP secretion is increased in obesity, 2) the postprandial hyperinsulinemia of obesity is not due to excessive insulin secretion but is likely secondary to altered insulin clearance, and 3) GIP cannot account for the hyperinsulinemia of obesity through its insulinotropic action.
Microtubule and plasmalemmal reorganization: acute response to estrogen
Authors: Szego C. M. , Sjostrand B. M. , Seeler B. J. , Baumer J. W. , Sjostrand F. S.
Published: December 2017

The acute ultrastructural effects of estrogen in endometrial epithelial cells were investigated by transmission electron microscopy (TEM), with special reference to the microtubule (MT) apparatus and the luminal surface. Ovariectomized rats anesthetized with pentobarbitol sodium were injected intravenously with estradiol-17 beta (E2 beta), 0.5 micrograms/100 g body wt. At intervals from approximately 30 s to 30 min thereafter, 70-80 nm cross sections of a uterine horn were prepared for TEM. In placebo controls, cytoplasmic MT were conspicuous in length and number, whereas only a minimal population of short microvilli (MV) was evident. In contrast, the specimens subjected to E2 beta for only 35 s showed a significant decrease in MT number and length, with virtually complete depletion of these organelles by approximately 80 s. Concomitantly, the luminal MV exhibited striking enhancement in length and density. Thereafter, these rapid and reciprocal alterations of MT and MV underwent inversion. Thus MT structures began to reappear within 2 min, increasing progressively so that by 30 min their numbers were again substantial, although lengths remained diminished. During the same interval, the initial surge of luminal MV gradually subsided, to near-control appearance by 30 min. These coordinate, reciprocal, and biphasic responses are consistent with biochemical evidences of abrupt membrane perturbation associated with interception of estrogen at its cellular targets. The resultant modification of the intracellular environment may contribute to limited reorganization of cellular architecture and propagation of the hormonal signal.
Calmodulin antagonist effects on GnRH and secretogogue-induced release of bovine LH
Authors: Kile J. P. , Amoss M. S.
Published: December 2017

A study was performed to determine the possible role of calmodulin (CaM) in regulating gonadotropin-releasing hormone (GnRH)-induced luteinizing hormone (LH) release from the bovine pituitary using three structurally unrelated calmodulin antagonists. Primary calf anterior pituitary cell cultures (3 X 10(5)/well) were treated with either LH secretogogue (GnRH, 100 ng/ml; A23187, 2.5 microM; theophylline, 1 mM; prostaglandin E2, 1 microM; estradiol, 25 ng/ml; or KCl, 25 mM; final concentrations) or secretogogue plus CaM inhibitor in 1 ml Hanks' balanced salt solution plus 10 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid for 6 h. Significant (P less than 0.01) inhibition of GnRH- and A23187-stimulated LH release was obtained with calmidazolium (CMZ; 1-10 nM) and N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7; 1-10 microM). Both CMZ (10 nM) and W-7 (10 microM) significantly reduced (P less than 0.05) LH release by all of the other agents tested as well. In contrast, trifluoperazine (TFP; 0.1-100 microM) had no effect against most of the secretogogues tested. These results suggest that GnRH-stimulated LH release is in part a Ca2+-CaM-dependent process and may implicate a common CaM-dependent mechanism for LH release in general. The data also demonstrate a marked dichotomy in response between the phenothiazine, TFP, and other CaM antagonists in the calf pituitary.
Effect of a thermogenic agent, BRL 26830A, on insulin receptors in obese mice
Authors: Rochet N. , Tanti J. F. , Gremeaux T. , Van Obberghen E. , Le Marchand-Brustel Y.
Published: December 2017

The effect of a new type of antidiabetic agent, BRL 26830A, has been tested in obese mice. Since this drug increases thermogenesis, insulin receptor binding and kinase activity were studied in brown adipose tissue and skeletal muscle of mice made obese by gold thioglucose. At 1 mg.kg-1.day-1, a 3-wk treatment normalized the glycemia and increased the uncoupling protein content of brown adipose tissue. The insulin receptor number and its associated kinase activity increased only in brown adipose tissue. At 2 mg.kg-1.day-1, additional effects, i.e., a 20% reduction in body weight and a normalization of insulin receptor number both in brown adipose tissue and in skeletal muscle, were observed. All those results were obtained even though hyperinsulinemia was not corrected. At the higher drug dosage, insulin receptor kinase activity evolved in direct proportion to the receptor number in brown adipose tissue. By contrast, in skeletal muscle, the receptor kinase activity toward exogenous substrates increased more than the receptor number, suggesting that the alteration of insulin receptor kinase activity previously reported in skeletal muscle of obese mice was partly reversed by BRL 26830A. None of these parameters was modified by the drug in lean mice. These results show that, even without affecting obesity, BRL 26830A improves insulin resistance in obese mice, probably through its effect on insulin receptors. This action prevails in brown adipose tissue, supporting the idea that this tissue plays an important role in glucose homeostasis. Thermogenic drugs could thus be powerful agents for the treatment of noninsulin-dependent diabetics.
Effects of repeated cycles of fasting-refeeding on brown adipose tissue composition in mice
Authors: Desautels M. , Dulos R. A.
Published: December 2017

Mice fasted for 24 h showed reductions in carcass fat and gonadal fat depots and atrophy of brown adipose tissue (BAT) that was characterized by loss of protein and succinate dehydrogenase. These changes were reversed on 24 h of refeeding. Cycling mice experienced 14 cycles of 1 day of fast followed by 2 days of refeeding, whereas control mice were fed ad libitum. Weight loss during each fast remained constant, and the animals lost and regained in excess of twice their initial weights within 6 wk. However, final weight and carcass and gonadal fat weights were similar to those of animals fed ad libitum. Total food intake was similar between cycling mice and those fed ad libitum suggesting an increase in feeding efficiency. There was no development of resistance to food deprivation since the preceding fasting experience of the animal had no effect on weight and carcass fat loss during a 24- or 48-h fast. Norepinephrine-stimulated oxygen consumption that was reduced in cycling mice was probably the result of a reduction of BAT thermogenic capacity. BAT succinate dehydrogenase content and the concentration of uncoupling protein in isolated mitochondria were significantly reduced. These changes in BAT composition were not observed when the refeeding period of each cycle was increased to 6 days. These results suggest that reduced energy expenditure in BAT may play a role in the conservation of energy during intermittent and frequent bouts of food deprivation.
Pancreatic islet discrimination of hexose anomers. I. Steady-state computer simulation
Authors: Achs M. J. , Garfinkel D.
Published: December 2017

Pancreatic islets detect glucose level by phosphorylating it and converting the glycolytic rate to a signal to secrete insulin. Insulin secretion is greater from the alpha- than from the beta-anomer when the D-glucose level is below 22 mM. D-mannose behaves similarly but at nearly twofold higher concentrations. Two explanations have been proposed: 1) glucokinase, which has the same anomeric preference, is the principal hexose phosphorylating enzyme and limits glycolytic rate. 2) Phosphofructokinase limits glycolysis and hexokinase is the principal enzyme phosphorylating hexose; hexosediphosphate activators of phosphofructokinase are more readily synthesized from alpha-anomers of hexose phosphates. We have simulated both alternatives with a detailed anomerically specific model of the hexose-metabolizing glycolytic enzymes. The pathway preference for alpha-anomer of both hexoses was adequately reproduced with anomerically active limiting glucokinase. The other mechanism did not reproduce the observed pathway preference.
Steroid inhibition of canine ACTH: in vivo evidence for feedback at the corticotrope
Authors: Keller-Wood M. , Leeman E. , Shinsako J. , Dallman M. F.
Published: December 2017

We infused submaximal feedback doses of either dexamethasone (DEX; 0.1 microgram.kg-1.min-1) or corticosterone and cortisol (B+F; 1.5 micrograms.kg-1.min-1) intravenously for 40 min into conscious dogs and measured the adrenocorticotropic hormone (ACTH) responses to hypoglycemia induced by insulin (0.1 U/kg) or to ovine corticotropin-releasing factor (oCRF; 1 microgram/kg); both agents were injected at 120 min. The dose of DEX was chosen to produce suppression of the ACTH response to oCRF equivalent to that produced by B+F. The purpose of the study was to determine 1) whether CRF- and hypoglycemia-induced ACTH secretion are equally inhibited by glucocorticoid treatment and 2) whether DEX and B+F have differential effects in the inhibition of stress-induced ACTH secretion. We found that peak ACTH responses to hypoglycemia and CRF were equally inhibited by DEX (36 +/- 6 and 52 +/- 9%, respectively). The peak ACTH responses to hypoglycemia and CRF were also equally inhibited after B+F infusion (45 +/- 13 and 65 +/- 5%, respectively). There was no significant interaction between the steroid administered and the stimulus given in controlling the ACTH response (by 2-way analysis of variance). The results suggest that pituitary feedback is of primary importance in suppression of canine ACTH secretion by delayed feedback and that the natural and synthetic steroids both act at this site.
Effects of flow rate and insulin on triacylglycerol secretion by perfused rat liver
Authors: Topping D. L. , Storer G. B. , Trimble R. P.
Published: December 2017

In rat livers perfused with undiluted rat blood at perfusion rates of 6, 12, or 18 ml/min, hepatic O2 consumption rose with blood flow. Lipogenesis was unaffected by blood flow in control livers and was enhanced by insulin at 12 and 18 ml/min. Very-low-density lipoprotein triacylglycerol secretion also rose with increased flow and was stimulated by insulin at both 6 and 12 ml/min. When glucose was added to livers perfused at 12 or 18 ml/min, uptake was independent of perfusion rate and was slightly stimulated by insulin. Total lipogenesis and the secretion of newly synthesized fatty acids in very-low-density lipoprotein triacylglycerols were unaffected by insulin at either flow rate. The hormone stimulated triacylglycerol secretion at 18 ml/min but inhibited it at 12 ml/min. It seems that in perfused liver, effects of insulin on lipogenesis and very-low-density lipoprotein secretion may be modified not only by changes in O2 consumption (in this case through alterations in blood flow) but also by the choice of substrate.
Gonadal steroid effects on LH response to arachidonic acid and protein kinase C
Authors: Kamel F. , Kubajak C. L.
Published: December 2017

Cultured rat pituitary cells were used to examine, first, the effects of luteinizing hormone-releasing hormone, Ca2+ mobilization, protein kinase C (PKC) activation, and arachidonic acid (AA) on luteinizing hormone (LH) secretion and AA release, and, second, gonadal steroid modulation of these effects. A23187, 12-O-tetradecanoylphorbol 13-acetate (TPA), and AA stimulated LH secretion by both perifused and static cultures; TPA facilitated the responses to both A23187 and AA. LHRH, A23187, and TPA stimulated AA release. Inhibition of AA metabolism reduced the LH responses to LHRH, A23187, TPA, and melittin. Pretreatment with testosterone inhibited the LH response to LHRH but not the responses to TPA or AA. Pretreatment with 17 beta-estradiol stimulated the LH responses to LHRH, TPA, and low concentrations of AA. These results suggest that LHRH action involves a cascade of events, in which the effects of Ca2+ mobilization and PKC activation are mediated at least in part by AA release. They further suggest that both testosterone and 17 beta-estradiol modulate LH secretion by affecting AA release; 17 beta-estradiol may also affect some process subsequent to AA release.
Metabolic role of the exercise-induced increment in epinephrine in the dog
Authors: Moates J. M. , Lacy D. B. , Goldstein R. E. , Cherrington A. D. , Wasserman D. H.
Published: December 2017

The role of the exercise-induced increment in epinephrine was studied in five adrenalectomized (ADX) and in six normal dogs (C). Experiments consisted of an 80-min equilibration period, a 40-min basal period, and a 150-min exercise period. ADX were studied with epinephrine replaced to basal levels during rest and to increased levels during exercise to simulate its normal rise (HE) and on a separate day with epinephrine maintained at basal levels throughout the study (BE). Cortisol was replaced during rest and exercise in ADX so as to simulate the levels seen in C. Glucose was infused as needed in ADX to maintain the glycemia evident during exercise in C. Glucose production (Ra) and utilization (Rd) were assessed isotopically. In C, epinephrine had risen by 95 +/- 25 pg/ml by the end of exercise. In HE, the increment in epinephrine (117 +/- 29 pg/ml) was similar to that seen in C, whereas in BE epinephrine fell by 18 +/- 9 pg/ml. Basal norepinephrine levels were 139 +/- 9, 260 +/- 25, and 313 +/- 33 pg/ml in C, HE, and BE, respectively. In response to exercise, norepinephrine increased by nearly twofold in all protocols. Basal and exercise-induced changes in plasma glucagon and insulin were similar in C and ADX. Ra increased similarly in C (5.3 +/- 0.6 mg.kg-1.min-1) and HE (4.9 +/- 0.6 mg.kg-1.min-1). In BE, Ra rose normally for the initial 90 min but then declined resulting in a rise of only 2.9 +/- 0.5 mg.kg-1.min-1 after 150 min of exercise.(ABSTRACT TRUNCATED AT 250 WORDS)