bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of reciprocal pool specific activities to model leucine metabolism in humans
Authors: Schwenk W. F. , Beaufrere B. , Haymond M. W.
Published: December 2017

To investigate in humans whether interconversion of leucine and its alpha-keto acid, alpha-ketoisocaproate (KIC), is rate limiting for leucine metabolism, normal volunteers were infused with independent 3H and 14C labels of leucine and KIC. Total leucine carbon (leucine + KIC) entry into the plasma space determined by using both plasma specific activities (SA) of the infused isotopes or by using either plasma SA of the leucine moieties reciprocal to those being infused (e.g., [3H]KIC and [14C]leucine during [3H]leucine and [14C]KIC infusion) were nearly identical. When [3H]leucine and [14C]KIC were infused, the ratio of 3H/14C radioactivity of leucine in plasma proteins was similar both to the ratio of plasma [3H]KIC SA/[14C]leucine SA and to the ratio of the rate of [3H]leucine/[14C]KIC infused. Estimates of leucine oxidation using expired 14CO2 and the plasma SA of the 14C moiety reciprocal to infused [14C]leucine or [14C]KIC were similar. We conclude that the plasma SA of the leucine moiety reciprocal to infused labeled leucine or KIC may reflect more accurately whole-body leucine metabolism in postabsorptive humans.
Glucose metabolism in epitrochlearis muscle of acutely exercised and trained rats
Authors: Davis T. A. , Klahr S. , Tegtmeyer E. D. , Osborne D. F. , Howard T. L. , Karl I. E.
Published: December 2017

Effects of insulin on glycogen synthesis (GS), glycolytic utilization (GU), and glucose uptake (GT) were studied in isolated epitrochlearis muscles from exercise-trained or sedentary rats during recovery from acute exercise or at rest. During the 1st h after acute exercise, the enhanced basal and insulin-stimulated GT was directed mainly toward replenishment of glycogen but basal GU was also increased. During the second through third hours after exercise, basal GS decreased but remained greater than rest and basal GU and GT returned to normal. Insulin sensitivity of these parameters was enhanced. Training alone reduced basal GS but enhanced insulin sensitivity of GT and GU. Training reduced the acute exercise-stimulated increase in basal and insulin sensitivity of GS during recovery from acute exercise, probably due to elevated glycogen stores. Thus recovery from acute exercise or training, either alone or in combination, enhances insulin stimulated GT in muscle; however, the increased glucose is primarily channeled toward GS after acute exercise, which is reduced by prior training and is directed to GU in trained animals either at rest or after acute exercise.
Endotoxin treatment inhibits glucocorticoid induction of hepatic enzymes at a late induction step
Authors: Shackleford G. M. , Hart S. F. , Berry L. J.
Published: December 2017

Bacterial endotoxin inhibits the glucocorticoid induction of several hepatic enzymes including phosphoenolpyruvate carboxykinase (PEPCK). Experiments were performed to elucidate the mechanism of this inhibition by examination of the early events in the glucocorticoid induction process. At a dose of endotoxin 2-to 10-fold greater than that required to inhibit the induction of PEPCK activity, no effect on the entry of glucocorticoids into hepatocytes or their ability to form complexes with glucocorticoid receptors could be measured. Binding data showed no effect of endotoxin treatment on the association or dissociation kinetics of the steroid-receptor binding reaction. Scatchard analysis revealed no effect on the affinity and number of hepatic glucocorticoid receptor binding sites, indicating that down-regulation of receptors is not responsible for inhibition of induction. Finally, activation of receptor complexes was unaffected as well by endotoxin treatment. We conclude from these data that endotoxin does not act at the early events in the glucocorticoid induction process and must therefore intervene at a subsequent step.
Estrogen inhibition of LH and FSH secretion: effects of a GnRH antagonist
Authors: Charlesworth M. C. , Schwartz N. B.
Published: December 2017

Follicle-stimulating hormone (FSH) levels are not suppressed as rapidly or to the same degree as luteinizing hormone (LH) levels in ovariectomized rats treated with either gonadotropin-releasing hormone (GnRH) antagonist or estrogen. The acute inhibitory effects of various doses of estrogen on FSH and LH secretion were examined in cannulated, 2-wk ovariectomized rats. No dose of 17 beta-estradiol, up to 2,500 ng injected intravenously, suppressed FSH, although LH secretion was inhibited 50% within 1 h by 100 ng. In another experiment, estradiol benzoate (EB; 10 or 250 micrograms; sc injection in oil) was only marginally effective in suppressing FSH, compared with LH, levels in serum. Treatment with EB 24 h before or after a 500 micrograms dose of a GnRH antagonist did not reduce LH or FSH to levels lower than those achieved with antagonist alone. These results indicate that the GnRH-dependent component of FSH release and the GnRH-independent component that is unmasked in the presence of GnRH antagonist are sensitive to negative feedback by estrogen, indicating that this steroid is not the primary inhibitory ovarian factor regulating FSH in the rat.
Thermogenesis and sympathetic activity in BAT of overfed rats after adrenalectomy
Authors: Marchington D. , Rothwell N. J. , Stock M. J. , York D. A.
Published: December 2017

Resting oxygen consumption was elevated by 30% in young rats fed a cafeteria diet compared with their chow-fed controls and by 22% in cafeteria-fed, adrenalectomized (ADX) rats compared with the ADX chow-fed group, but injection of propranolol reduced oxygen consumption in the cafeteria-fed animals and abolished these differences. Brown adipose tissue (BAT) mass was increased by cafeteria feeding, and the activity of the mitochondrial proton conductance pathway (assessed from purine nucleotide binding) was enhanced by adrenalectomy and by cafeteria feeding. Norepinephrine turnover in BAT (determined from the time-dependent loss of tissue [3H]norepinephrine specific activity) was increased by 105% in sham-operated, cafeteria-fed rats, by 142% in chow-fed ADX rats, and by 400% in cafeteria-fed ADX rats, compared with chow-fed controls. Cardiac norepinephrine turnover was elevated by 80% in sham-operated, cafeteria-fed rats, but unaffected by adrenalectomy. These data indicate that the enhanced thermogenesis and BAT activity induced by adrenalectomy in chow- or cafeteria-fed rats is due to increased sympathetic activity in the tissue.
Glucoregulatory role of cortisol and epinephrine interactions studied in adrenalectomized dogs
Authors: Gauthier C. , el-Tayeb K. , Vranic M. , Lickley H. L.
Published: December 2017

The importance of basal cortisol (H) and epinephrine (E) levels on glucoregulation, and the effects of E, given to simulate moderate to severe stress (5 times basal rate of infusion), were examined in seven conscious adrenalectomized dogs. Although plasma glucagon (IRG) increased by 47%, insulin (IRI) decreased by 36%, norepinephrine (NE) increased by 103%, and FFA decreased by 26%, glucose concentration and kinetics remained normal after adrenalectomy. A 4-h infusion of H reestablished basal cortisol levels and returned IRG to its basal preadrenalectomy level with no change in IRI, NE, and FFA levels. Glucose production and metabolic clearance decreased concomitantly by 20%, maintaining euglycemia. A 90-min infusion of basal E caused only a transient increase in IRG. The simultaneous infusion of H with E prevented this increase in IRG and returned IRI to preadrenalectomy levels in the absence of any change in NE or glucose. A subsequent infusion of five times basal E, alone, raised circulating E levels and caused a transient decrease in plasma NE, but no change in IRI. There was a similar hyperglycemic response, as seen previously in normal dogs. The simultaneous infusion of H and E prevented the decrease in NE, but did not change the IRI and FFA responses. There was an 80% greater plasma glucose response than seen during infusion of E alone. In conclusion, what E and H lack after adrenalectomy is compensated for by an increase in IRG and a decrease in IRI, and normal glucose concentrations and kinetics are maintained. It appeared that normoglucagonemia required basal H release, whereas normoinsulinemia required both basal H and E secretion.(ABSTRACT TRUNCATED AT 250 WORDS)
Blood flow distribution in tissues of perfused rat hindlimb preparations
Authors: Gorski J. , Hood D. A. , Terjung R. L.
Published: December 2017

Aerobic muscle metabolism during concentrations requires adequate blood flow and oxygen delivery. Since the perfused rat hindquarter (HQ) has become widely used for muscle stimulation, we examined the blood flow distribution, using 15 microns radiolabeled microspheres, and oxygen consumption of the HQ, using different commonly used perfusion protocols. Perfusion via the abdominal aorta resulted in well-matched (r = 0.90) blood flows between tissues of both hindlimbs that were proportional to total perfusion inflow. Blood flows to the high-oxidative fast-twitch and slow-twitch red muscle sections were three- to fourfold greater than flows to sections of low-oxidative fast-twitch white muscle. However, a large fraction (28%) of the total inflow went to the trunk region, even though all apparent arterial branches to the trunk region were ligated. This trunk mass accounts for at least 40% of the total metabolic responses of the HQ and diverts a large blood flow that is often presumed to supply the hindlimbs. As a result, muscle performance of the distal hindlimb muscle during stimulation can be inordinately poor. Ligation of the iliac artery to the contralateral limb improves blood flow to the remaining hindlimb but does not eliminate trunk blood flow. In contrast, perfusion via the femoral artery restricted 95% of the inflow to the single hindlimb, thereby reducing the tissue mass perfused. Blood flow to the distal limb musculature was high, resulting in an enhanced muscle performance. Thus single hindlimb perfusion provides a preparation where the contracting muscle is a large fraction of the total tissue, and the venous effluent better reflects the metabolic events in the contracting muscle.(ABSTRACT TRUNCATED AT 250 WORDS)
TRH and GRF stimulate release of growth hormone through different mechanisms
Authors: Szabo M.
Published: December 2017

Thyrotropin-releasing hormone (TRH) is an effective stimulator of growth hormone (GH) release from cultured adenohypophysial cells of chronically hypothyroid rats in vitro. The present study explored the question of cAMP and calcium mediation of the GH-stimulatory effect of TRH in this system. A maximally stimulatory concentration of TRH was added together with various concentrations of human GH-releasing factor 40 (hGRF-40) whose action is cAMP mediated, or of dibutyryl cAMP (DBcAMP), to primary monolayer cultures of adenohypophysial cells from thyroidectomized rats. The GH responses to the combined addition of TRH with all doses of GRF or DBcAMP were fully additive, causing parallel elevations of the dose-response curves. Whereas the GH response to maximally effective concentrations of hGRF-40 and DBcAMP, added together, was not greater than that to either secretagogue alone, the inclusion of TRH increased the response to a new Emax. The calcium inhibitors, verapamil, EGTA, and CoCl2, markedly suppressed basal GH release and virtually completely blocked the GH response to TRH, suggesting calcium mediation. In chronically hypothyroid, urethan-anesthetized rats, the in vivo effect of the combined administration of maximally effective doses of TRH and GRF on plasma GH levels was also additive. These findings indicate that TRH stimulates GH release in adenohypophysial cells of hypothyroid rats by a cAMP-independent, calcium-dependent mechanism.
Synergistic improvement of glucose tolerance by sucrose feeding and exercise training
Authors: Vallerand A. L. , Lupien J. , Bukowiecki L. J.
Published: December 2017

The interactive effects of exercise training (5-7 wk) and sucrose consumption (ad libitum feeding of a 32% sucrose solution and Purina chow) on intravenous glucose tolerance and plasma insulin levels were investigated using a 2 X 2 experimental design. Rats were divided in Purina-sedentary, Purina-trained, sucrose-sedentary, and sucrose-trained groups. Sucrose feeding of sedentary animals significantly increased basal and glucose-stimulated insulin levels and improved basal glycemia and glucose tolerance. On the other hand, exercise training of Purina-fed animals significantly reduced basal as well as glucose-stimulated insulinemia without altering basal glycemia or glucose tolerance. Such a sparing effect of exercise training on insulin requirements was not as evident in rats consuming sucrose. These animals displayed a reduced basal glycemia (P less than 0.01) with normal basal insulin levels. Their glucose tolerance was markedly improved (P less than 0.01) but their insulin response during intravenous glucose tolerance test remained as high as in sucrose-sedentary animals. Results from these studies indicate that sucrose feeding of sedentary animals leads to hyperinsulinemia without compensatory insulin resistance, resulting in an improvement of glucose tolerance, exercise training increases the sensitivity of peripheral tissues to insulin, and the marked improvement of glucose tolerance observed in sucrose-trained animals results from a synergistic combination of the above two factors, i.e., increased insulinemia (induced by diet) and enhanced insulin sensitivity (induced by training).
Sex-related differences in GH secretion in rat using reverse hemolytic plaque assay
Authors: Ho K. Y. , Leong D. A. , Sinha Y. N. , Johnson M. L. , Evans W. S. , Thorner M. O.
Published: December 2017

It is not known whether enhanced growth hormone (GH)-releasing factor (GRF)-stimulated GH release observed in the male reflects differences in somatotrope numbers and/or secretory response to GRF. We addressed this question by using the hemolytic plaque assay which allows quantification of hormone secretion by single pituitary cells. Time-course studies and GRF-GH concentration-response relationships (0.01, 0.1, 1, 10, 100, 1,000 nM GRF) in age-matched male and diestrous day 2 female rats were compared by quantitating the percent of GH plaque-forming cells, and measuring the plaque areas. The male pituitary contained a greater percent (P less than 0.05) of somatotropes (% of plaque-forming cells 45 +/- 2 vs. 27 +/- 4% in the female; mean +/- SE). GRF induced a greater concentration-dependent increase in plaque areas in the male. Maximal responses were attained at 10 nM GRF in both sexes. However, mean maximal plaque area was significantly greater (P less than 0.001) and the EC50 was significantly lower (P less than 0.05) in the male (0.25 +/- 0.09 vs. 1.78 +/- 0.64 nM in the female). The data suggest that the greater percent of somatotropes in the male and greater secretory capacity and sensitivity to GRF may contribute to sex-related differences in GH secretion in the rat.
Role of vitamin D in neonatal skeletal development in rats
Authors: Mathews C. H. , Brommage R. , DeLuca H. F.
Published: December 2017

The role of vitamin D in rat pup growth and skeletal development without the influence of nutritional factors was investigated. Pups from vitamin D-replete and vitamin D-deficient dams receiving identical amounts of milk for 20 days were compared. Body weight gain, femur ash content and histomorphometric analyses of diaphysial and distal femur were determined. Up to 20 days of age, growth and skeletal development of the pups were completely normal in the absence of vitamin D. Skeletal changes found in vitamin D deficiency were not observed, i.e., there was no increased volume of osteoid or lack of bone mineralization as demonstrated by tetracycline labeling and ash content. Only increased cortical porosity was found in vitamin D-deficient pups. Therefore, abnormalities previously attributed to vitamin D deficiency in neonatal rats can be corrected by sufficient milk consumption and are thus not a direct function of vitamin D.
Binding of IGF I and IGF I-stimulated phosphorylation in canine renal basolateral membranes
Authors: Hammerman M. R. , Gavin J. R.
Published: December 2017

To characterize the interaction of the renal proximal tubular cell with insulin like growth factor I (IGF I), we measured binding of 125I-IGF I to proximal tubular basolateral membranes from dog kidney and induced IGF I-stimulated phosphorylation of basolateral membranes. Specific binding of 125I-IGF I to basolateral membranes was observed that was half-maximal at between 10(-9) and 10(-8) M IGF I. 125I-IGF I was affinity cross-linked to a 135,000 Mr protein in basolateral membranes that was distinct from the alpha-subunit of the insulin receptor and from the IGF II receptor. IGF I-stimulated phosphorylation of a 92,000 Mr protein was effected in detergent-solubilized membranes incubated with 100 microM [gamma-32P]ATP. The 32P-labeled protein was distinct from the beta-subunit of the insulin receptor, the 32P phosphorylation of which was stimulated by insulin. We conclude that specific receptors for IGF I are present in the basolateral membrane of the renal proximal tubular cell. Physiological actions of IGF I at this nephron site may occur through the binding of this peptide circulating in plasma, to specific basolateral membrane receptors, followed by IGF I stimulated phosphorylation.
Peptides PHI and VIP: comparison between vascular and nonvascular smooth muscle effect in rabbit uterus
Authors: Bardrum B. , Ottesen B. , Fahrenkrug J.
Published: December 2017

The distribution and effects of the two neuropeptides, vasoactive intestinal polypeptide (VIP) and peptide histidine isoleucine amide (PHI), on vascular and nonvascular smooth muscle in the urogenital tract of nonpregnant rabbit female, were investigated. Immunoreactive VIP and PHI were present in all regions except the ovary with the highest concentration in the uterine cervix. By using in vitro tension recordings of myometrial specimens, it was demonstrated that both peptides displayed a dose-dependent inhibition of the mechanical activity. The dose-response curves of VIP and PHI were superimposable with and ID50 of 3 X 10(-8) mol/l, and their combined effect was additive. In addition, the influence of the two peptides on myometrial blood flow (MBF) was investigated by the xenon-133 washout technique. Both peptides were found to increase MBF with the same potency and efficacy. Their combined effect was additive. In conclusion VIP and PHI are present in the rabbit urogenital tract, and the two peptides are equipotent inhibitors of mechanical nonvascular and vascular smooth muscle activity in the uterus.
Chronic exercise increases insulin binding in muscles but not liver
Authors: Bonen A. , Clune P. A. , Tan M. H.
Published: December 2017

It has been postulated that the improved glucose tolerance provoked by chronic exercise is primarily attributable to increased insulin binding in skeletal muscle. Therefore, we investigated the effects of progressively increased training (6 wk) on insulin binding by five hindlimb skeletal muscles and in liver. In the trained animals serum insulin levels at rest were lower either in a fed (P less than 0.05) or fasted (P less than 0.05) state and after an oral glucose tolerance test (n = 8) (P less than 0.05). Twenty-four hours after the last exercise bout sections of the liver, soleus (S), plantaris (P), extensor digitorum longus (EDL), and red (RG) and white gastrocnemius (WG) muscles were pooled from four to six rats. From control animals, killed at the same time of day, muscles and liver were also obtained. Insulin binding to plasma membranes increased in S, P, and EDL (P less than 0.05) but not in WG (P = 0.07), RG (P greater than 0.1), or in liver (P greater than 0.1). There were insulin binding differences among muscles (P less than 0.05). Comparison of rank orders of insulin binding data with published glucose transport data for the same muscles revealed that these parameters do not correspond well. In conclusion, insulin binding to muscle is shown to be heterogeneous and training can increase insulin binding to selected muscles but not liver.
Effects of thyroid hormones on calcium contents and 45Ca exchange in rat skeletal muscle
Authors: Everts M. E. , Clausen T.
Published: December 2017

In 4-wk-old rats, pretreatment with L-triiodothyronine (T3) increased calcium content by 100% and the 30-min 45Ca uptake by 64% in the soleus, whereas the extensor digitorum longus (EDL) muscle showed no significant change. The stimulation of 45Ca uptake was resistant to dantrolene and methoxyverapamil (D600) and could not be attributed to altered permeability of the plasma membrane to calcium, but appears to reflect increased net accumulation of calcium in intracellular pools. The stimulating effect of high K0 (20 mM) on 45Ca uptake was more pronounced in soleus than in EDL and could be suppressed by dantrolene and D600. The results indicate that the effects of T3 on calcium content and 45Ca exchange are primarily exerted on muscles containing a large proportion of slow-twitch, oxidative fibers. In soleus muscle from hyperthyroid rats the stimulating effects of high K0 on 45Ca uptake and lactate production were, respectively, 3.4 and 4.5 times larger than in those obtained from controls. These observations further support the earlier proposed idea [C. van Hardeveld and T. Clausen. Am. J. Physiol. 247 (Endocrinol. Metab. 10): E421-E430, 1984] that the metabolic effects of thyroid hormone depend on the availability of cellular as well as extracellular calcium.
Effect of pure zinc deficiency on glucose tolerance and insulin and glucagon levels
Authors: Park J. H. , Grandjean C. J. , Hart M. H. , Erdman S. H. , Pour P. , Vanderhoof J. A.
Published: December 2017

The effect of zinc deficiency on glucose tolerance was investigated using intragastric force feeding to obviate decreased food intake and altered eating patterns. Three groups of weanling male Sprague-Dawley rats were fed a purified zinc-deficient diet: zinc-deficient, ad libitum-fed animals (ZDA) were offered powdered zinc-deficient diet; zinc-replete, force-fed controls (ZRF) were tube fed a diet blended with water containing 25 ppm of zinc; zinc-deficient, force-fed animals (ZDF) were similarly tube fed the zinc-deficient diet. The ZRF and ZDF groups received a diet of identical amount based on the intake of ad libitum-fed, zinc-replete rats. After 8 days of feeding, the ZDF group had impaired glucose tolerance curves, yet blood insulin and glucagon levels were normal. The ZDA group had normal glucose tolerance with low insulin levels compared with the ZRF group. The islet cell morphology among the three dietary groups were similar. These results suggest that the glucose intolerance observed in ZDF rats is not due to altered blood insulin and glucagon levels but rather to peripheral resistance to insulin action.
Renin heterogeneity in stroke-prone hypertensive and normotensive rats
Authors: Sessler F. M. , Jokelainen P. T. , Sing C. F. , Strack A. M. , Malvin R. L.
Published: December 2017

Six forms of renin are found in the rat kidney. We studied their secretion in renal slices from spontaneously hypertensive stroke-prone rats (SHRSP) and Wistar-Kyoto rats (WKY). Incubation media from renal slices were subjected to isoelectric focusing. Six peaks of renin activity with different isoelectric points were found. The renin concentration of each form was expressed as a percentage of the total recovered from the gel. We established that the forms secreted by renal slices of SHRSP differed from those of WKY: SHRSP slices released a higher proportion of forms focusing at the more acidic pH. The distribution of the six renin forms and of blood pressure (BP) among animals of the F1, F2, and backcross progenies resulting from the cross of SHRSP and WKY rats were studied. In the F1, BP, percentage of renin form 2, and a combination of the percentage of forms 4 + 5 + 6 were intermediate between the parental lines. The backcross rats showed BP and percentages of forms closer to their SHRSP or WKY parent. In the F2, the distribution of BP, percentage of forms 2 and 4 + 5 + 6 take the form of a unimodal distribution with a significantly larger variance than F1. The increase in the correlation between percentage of renin forms and BP, and between renin concentration of BP, in the segregating progenies over that observed in the parental lines and the F1, are support for the hypothesis that these traits are under the control of common genetic mechanisms.
Enhanced thermogenic response to a diet low but adequate in protein persists in older rats
Authors: Swick A. G. , Swick R. W.
Published: December 2017

Feeding a 5% lactalbumin diet to rats for 3 wk compared with a 15% lactalbumin diet resulted in lower feed efficiency and stimulated brown adipose tissue activity as measured by GDP binding. This occurred in 4-mo-old rats as well as in 7-mo-old rats. Furthermore the effects persisted after 3.5 mo of dietary treatment. An investigation of pattern of food intake revealed that no obvious alteration in pattern was involved in the lower feed efficiency associated with rats fed a 5% lactalbumin diet.
Effect of diet change on insulin action: difference between muscles and adipocytes
Authors: Maegawa H. , Kobayashi M. , Ishibashi O. , Takata Y. , Shigeta Y.
Published: December 2017

To investigate the difference of insulin action between skeletal muscle and adipose tissue in response to dietary manipulation, we studied the effect of high-sucrose (HS) and high-fat (HF) diet on insulin action by measuring insulin binding and insulin action both in soleus muscles and adipocytes. HS feeding led to a 14 and 28% decrease, and HF feeding led to a 25 and 36% decrease in insulin binding both to soleus muscles and adipocytes, respectively (P less than 0.01). In HF-fed rats, both rates of glucose uptake and intracellular glucose metabolism were impaired by 30-40% in soleus muscles (P less than 0.01), and adipocyte glucose uptake was also decreased by 30% in the submaximally insulin-stimulated state (P less than 0.05). On the other hand, in HS-fed rats with prominent hyperinsulinemia, glucose uptake was 2.3- to 2.7-fold increased in adipocytes (P less than 0.01). However, both rates of glucose uptake and intracellular glucose metabolism were not increased in soleus muscles from HS-fed rats. Steady-state plasma glucose (SSPG) level was unchanged in HS-fed rats during somatostatin, glucose, and insulin infusion, whereas the SSPG level of HF-fed rats was twice as much as that in controls (P less than 0.01). These results indicate that long-term regulation of glucose metabolism by ambient insulin in skeletal muscle may be different from that in adipocytes and that insulin action in muscle, rather than in adipocyte, may reflect insulin action of whole body.
Clenbuterol, a beta 2-agonist, retards atrophy in denervated muscles
Authors: Zeman R. J. , Ludemann R. , Etlinger J. D.
Published: December 2017

Denervated soleus, anterior tibialis, and gastrocnemius muscles, but not the extensor digitorum longus, contained 95–110% more protein after 2–3 wk of treatment with the adrenergic beta 2-receptor agonist, clenbuterol, than denervated controls. In addition, the twofold difference in the protein content of denervated solei was paralleled by similar changes in contractile strength and muscle fiber cross-sectional area. In contrast, when the innervated contralateral muscles were examined, the extensor digitorum longus and anterior tibialis showed relatively small increases in protein of 32 and 19%, respectively, whereas the soleus and gastrocnemius were unaffected. The magnitude of the effects of clenbuterol in sparing the mass and functional capacity of denervated muscle suggests that this agent may be important in studies of neuromuscular diseases and disuse atrophy.
Effects of forskolin on bone in organ culture
Authors: Krieger N. S. , Stern P. H.
Published: December 2017

The effects of forskolin, which directly activates adenylate cyclase in most systems, have been compared with the actions of parathyroid hormone and calcitonin, both of which have been suggested to utilize cAMP as a second messenger in their actions on bone. Forskolin alone stimulated calcium release from neonatal mouse calvaria and fetal rat limb bones in vitro in a dose-dependent manner. The effect was maximal at 10(-6) M in both systems. At higher concentrations forskolin completely inhibited stimulated bone resorption, although with submaximal concentrations the inhibition was only partially sustained up to 72 h. Forskolin directly stimulated cAMP release from calvaria into the medium at concentrations up to 10(-4) M. Forskolin had no effect on the interaction between parathyroid hormone and calcitonin, while calcitonin inhibited the stimulatory effect of forskolin comparably with its inhibition of parathyroid hormone-stimulated bone resorption. The results indicate that forskolin has dual effects on bone and can mimic responses of both parathyroid hormone and calcitonin in both bone culture systems. The observed response depends on the concentration of forskolin used and the length of treatment with the drug.
Estrogen effects on angiotensin receptors are modulated by pituitary in female rats
Authors: Douglas J. G.
Published: December 2017

The present studies were designed to test the hypothesis that changes in angiotensin II (ANG II) receptors might modulate the altered target tissue responsiveness accompanying estradiol administration. Estradiol was infused continuously in oophorectomized female rats by employing minipumps to achieve plasma estradiol levels and simulating pregnancy levels in the rat. Aldosterone was also infused in control and experimental animals to avoid estrogen-induced changes in renin and ANG II. ANG II binding constants were determined in radioreceptor assays. Estradiol increased binding site concentration in adrenal glomerulosa by 76% and decreased binding sites of uterine myometrium and glomeruli by 45 and 24%, respectively. There was an accompanying increase in the affinity of ANG II binding to adrenal glomerulosa and uterine myometrium. Because estrogen is a potent stimulus of prolactin release from the pituitary of rodents, studies were also designed to test the hypothesis that prolactin may mediate some or all of the estrogen-induced effects observed. Hypophysectomy abolished estradiol stimulation of prolactin release and most ANG II receptor changes. The only effect that persisted was a 41% decrease in the density of uterine receptors. Prolactin administration to pituitary intact rats was associated with a 50% increase in receptor density of adrenal glomerulosa simulating estradiol administration. However, the changes in glomeruli and uterine myometrium were opposite in that both tissues also increased receptor density, suggesting that prolactin was not the sole mediator of the estrogen-induced receptor changes. In conclusion, regulation of ANG II receptors in a number of diverse target tissues by estradiol is complex with contributions from estrogens and pituitary factors, which include but do not exclusively involve prolactin.
Maitotoxin stimulates steroid and peptide hormone secretion by swine luteal tissue
Authors: Veldhuis J. D. , Yoshida K. , DuBois W. , Fields M. J.
Published: December 2017

The impact of calcium influx on the secretion of steroidal and nonsteroidal ovarian products was tested using a novel marine toxin, maitotoxin, a putative activator of endogenous calcium channels. In incubations of swine luteal minces in balanced salt solution, maitotoxin induced dose-dependent (3–12.5 ng/ml) and time-dependent (15–60 min) secretion of progesterone and relaxin. The magnitude of these stimulatory effects approached that observed for luteinizing hormone (LH) and was explicitly dependent on the presence of extracellular calcium ions. Maitotoxin's actions were not significantly impeded by substitution of choline for sodium ions in the incubation medium. Moreover, measurements of tissue calcium content by atomic absorption spectrometry indicated that maitotoxin augmented the total cellular content of calcium by 44 +/- 3.1% within 1 min. This time course was distinct from that observed for LH. The specificity of these actions on the calcium pathway was further suggested by the absence of any measurable effects of maitotoxin on luteal cAMP generation. We conclude that maitotoxin transiently increases luteal calcium content without altering cAMP generation and elicits concomitant release of both a steroidal (progesterone) and nonsteroidal (relaxin) product of the corpus luteum. Such actions are dependent on extracellular calcium but not sodium ions. These observations provide evidence for a calcium-dependent stimulatory pathway that activates both steroidal and nonsteroidal mechanisms of secretion in the swine corpus luteum.
WR-2721 inhibits parathyroid adenylate cyclase
Authors: Weaver M. E. , Morrissey J. , McConkey C. , Goldfarb S. , Slatopolsky E. , Martin K. J.
Published: December 2017

WR-2721 [S-2-(3-aminopropylamino)ethylphosphorothioic acid] is a chemoprotective and radioprotective agent that has been shown to lower serum calcium in dogs and in humans. This is secondary both to impaired release of Ca2+ from bone and diminished secretion of parathyroid hormone (PTH) from parathyroid glands. Because cAMP plays a role in the regulation of PTH secretion and WR-2721 has been shown to lower cAMP levels in radiated mouse spleen, we investigated the effects of WR-2721 on cAMP production in dispersed bovine parathyroid cells. Additionally, we studied the adenylate cyclase in plasma membranes from normal bovine parathyroid glands after exposure to WR-2721. With parathyroid cells incubated at 0.5 mM Ca2+, addition of WR-2721 in concentrations ranging from 0.02 to 2.0 mM resulted in a progressive decrease in intracellular cAMP (42-50%, respectively). In plasma membranes of bovine parathyroid cells a dose-dependent decrease in adenylate cyclase activity was noted. Inhibition of the cyclase was seen over a wide range of Mg2+ concentrations (2.5-40 mM). WR-2721 inhibited both basal and NaF, Gpp(NH)p, forskolin, and pertussin toxin-stimulated adenylate cyclase. These data suggest that WR-2721 inhibits the activity of parathyroid adenylate cyclase.
Polymyxin B selectively inhibits insulin effects on transport in isolated muscle
Authors: Gremeaux T. , Tanti J. F. , Van Obberghen E. , Le Marchand-Brustel Y.
Published: December 2017

Polymyxin B (PMB), a cyclic decapeptide antibiotic, inhibits the hypoglycemic effect of insulin in vivo. To elucidate the mechanism of PMB action, we have studied its effect in vitro on insulin-stimulated pathways in the mouse skeletal muscle. PMB, added to the incubation mixture, specifically inhibited insulin-stimulated 2-deoxyglucose transport and alpha-aminoisobutyric acid uptake in the isolated soleus muscle but did not affect the basal rates of transport (measured in the absence of insulin). PMB did not alter insulin binding and hexokinase activity. PMB effect was observed at all deoxyglucose concentrations tested, and PMB was also able to inhibit vanadate-stimulated glucose transport. By contrast, insulin activation of glycogen synthase was not prevented by PMB. Basal and maximally insulin-stimulated insulin receptor tyrosine kinase activity, tested in a cell-free system, was similar for both autophosphorylation and phosphorylation of exogenous substrates in the absence or in the presence of PMB. Furthermore, the insulin sensitivity of the kinase was increased in the presence of PMB. Our results suggest that the anti-insulin effect of PMB observed in vivo is due to an inhibition of insulin-stimulated glucose transport in the skeletal muscle perhaps through a specific blockade of the insulin-induced translocation of the glucose carriers.
Atrial natriuretic factor in maternal and fetal sheep
Authors: Cheung C. Y. , Gibbs D. M. , Brace R. A.
Published: December 2017

To determine atrial natriuretic factor (ANF) concentrations in the circulation and body fluids of adult pregnant sheep and their fetuses, pregnant ewes were anesthetized with pentobarbital sodium, and the fetuses were exteriorized for sampling. ANF concentration, as measured by radioimmunoassay, was 47 +/- 6 (SE) pg/ml in maternal plasma, which was significantly higher than the 15 +/- 3 pg/ml in maternal urine. In the fetus, plasma ANF concentration was 265 +/- 49 pg/ml, 5.6 times that in maternal plasma. No umbilical arterial and venous difference in ANF concentration was observed. Fetal urine ANF concentration (13 +/- 2 pg/ml) was significantly lower than that in fetal plasma, and was similar to that measured in amniotic and allantoic fluid. In chronically catheterized maternal and fetal sheep, fetal plasma ANF was again 5.1 times that in maternal plasma, and these levels were not different from those measured in acutely anesthetized animals. These results demonstrate that immunoreactive ANF is present in the fetal circulation at levels higher than those found in the mother. The low concentration of ANF in fetal urine suggests that ANF is probably metabolized and/or reabsorbed by the fetal kidney.
Metabolic basis of diethylaminoethoxyhexestrol-induced phospholipid fatty liver
Authors: Kubo M. , Hostetler K. Y.
Published: December 2017

Diethylaminoethoxyhexestrol caused a foam cell lipidosis in humans characterized by phospholipid storage in the liver, spleen, and other tissues, and this represents the first description of acquired lipidosis caused by a drug. It has been proposed that diethylaminoethoxyhexestrol causes phospholipid fatty liver by concentrating in lysosomes and inhibiting phospholipases but it has not previously been possible to measure the intralysosomal concentration of diethylaminoethoxyhexestrol. In this paper we report for the first time the intralysosomal concentration of this drug in rats. After a single oral dose of diethylaminoethoxyhexestrol (100 mg/kg) the intralysosomal concentration was 7.9 mM at 2.5 h, 15.6 mM at 12 h, and 20.9 mM at 24 h, respectively. The total phospholipid content of lysosomes in drug-treated rats increased 1.9-, 6.0-, and 7.6-fold over control at 2.5, 12, and 24 h, respectively. Purified lysosomal phospholipase A1 was strongly inhibited by diethylaminoethoxyhexestrol in vitro. In phospholipid fatty liver, phospholipid accumulation in lysosomes appears to be caused by the presence of diethylaminoethoxyhexestrol in lysosomes at concentrations estimated to be 7.9–20 mM, because drug levels above 1 mM completely block the activity of purified lysosomal phospholipase A1 in vitro.
Validation of a new method for determination of free fatty acid turnover
Authors: Miles J. M. , Ellman M. G. , McClean K. L. , Jensen M. D.
Published: December 2017

The accuracy of tracer methods for estimating free fatty acid (FFA) rate of appearance (Ra), either under steady-state conditions or under non-steady-state conditions, has not been previously investigated. In the present study, endogenous lipolysis (traced with 14C palmitate) was suppressed in six mongrel dogs with a high-carbohydrate meal 10 h before the experiment, together with infusions of glucose, propranolol, and nicotinic acid during the experimental period. Both steady-state and non-steady-state equations were used to determine oleate Ra ([3H]oleate) before, during, and after a stepwise infusion of an oleic acid emulsion. Palmitate Ra did not change during the experiment. Steady-state equations gave the best estimates of oleate inflow approximately 93% of the known oleate infusion rate overall, while errors in tracer estimates of inflow were obtained when non-steady-state equations were used. The metabolic clearance rate of oleate was inversely related to plasma concentration (P less than 0.01). In conclusion, accurate estimates of FFA inflow were obtained when steady-state equations were used, even under conditions of abrupt and recent changes in Ra. Non-steady-state equations, in contrast, may provide erroneous estimates of inflow. The decrease in metabolic clearance rate during exogenous infusion of oleate suggests that FFA transport may follow second-order kinetics.
Deprivation of corticosterone does not prevent onset of obesity in Zucker fa/fa pups
Authors: Bazin R. , Planche E. , Dupuy F. , Krief S. , Lavau M.
Published: December 2017

Adrenalectomy has been shown to reduce the development of obesity in adult Zucker fatty rat. In this study, we examined whether adrenalectomy could prevent the emergence of obesity and correct any of the first abnormalities to develop in fa/fa pups. Four-day-old Zucker pups were adrenalectomized and fed by adrenalectomized wet nurses until 11 days of age. The frequency distribution curves of fat cell volume clustered in two groups as they do in control litters, providing evidence that two phenotypes were present. Oxygen consumption measured at 8 days of age was significantly lower in fa/fa than in Fa/fa. Adrenalectomy did not restore the decreased oxygen consumption of fa/fa. In control litters, the GDP binding to brown adipose tissue mitochondria was twofold lower, whereas fatty acid synthase activity of this tissue was significantly increased in fa/fa pups. In inguinal adipose tissue of fa/fa pups, fatty acid synthase, and lipoprotein lipase activities were twice as active as in the tissue of lean pups. In adrenalectomized fa/fa pups, none of these metabolic abnormalities was corrected. The results demonstrate that adrenalectomy early in life did not prevent the emergence of obesity in suckling fa/fa rats.
Sexual dimorphism in adrenergic regulation of hepatic glycogenolysis
Authors: Studer R. K.
Published: December 2017

The total phosphorylase a plus b of hepatocytes isolated from females and incubated in the absence or presence of estradiol and progesterone at concentrations found in vivo does not vary during the estrous cycle. However, there is a slight but significant influence of the estrous cycle on basal and epinephrine-stimulated phosphorylase a activity, with a nadir being seen on diestrus. The relative contributions of the alpha- and beta-mediated pathways to phosphorylase a activation do not vary with the estrous cycle but are constant at 75 and 56%, respectively, of the response to 5 X 10(-8) M epinephrine. When the epinephrine-stimulated glucose release from glycogen stores in cells from females and males is compared, the release from the female is greater than that from the male, while the alpha-receptor-mediated stimulation in the female is comparable with that in the male. The beta-mediated pathway causes glucose release which averages 45% of that stimulated by epinephrine alone in cells from females. Basal cytosolic free calcium is similar in cells from males and females (142 vs. 149 nM, respectively). The epinephrine-stimulated increase in cytostolic free calcium (Cai) is greater in the male than the female at 10(-6) M (489 vs. 380 nM) but greater in the female than the male at 5 X 10(-9) M (54 vs. 27 nM). The changes in Cai are equivalent at intermediate epinephrine concentrations. When considered with our prior analysis of 45Ca efflux after adrenergic stimulation, this suggests there may be a sexual dimorphism in hepatocyte calcium transport systems. The glucose release for a given increase in Cai is greater in the female than the male probably due to the concomitant action of the beta-mediated increase in cAMP and the alpha-mediated increase in Cai. This supports the conclusion that the beta-mediated component does make a significant contribution to the catecholamine regulation of glycogenolysis in hepatocytes from adult female rats.
Effect of aldosterone on sodium and potassium concentrations in human mononuclear leukocytes
Authors: Wehling M. , Armanini D. , Strasser T. , Weber P. C.
Published: December 2017

The in vitro effect of aldosterone on intracellular sodium and potassium concentration was investigated in human mononuclear leukocytes (HML). HML were separated from blood by a Percoll gradient and intracellular Na+ and K+ were determined before and after incubation for 1 h at 37 degrees C. The intracellular (ic) Na+ and K+ concentrations after separation of HML were 17 +/- 5 and 59 +/- 18 mmol/kg wet cells (mean +/- SD, n = 6), respectively. In the absence of aldosterone the ic Na+ concentration decreased to 12 +/- 4, whereas the ic Na+ concentration remained constant at 18 +/- 8 mmol/kg wet cells when aldosterone (1.4 nM) was added to the incubation medium. In parallel, the ic K+ concentration decreased without added aldosterone but remained constant with aldosterone. The effect of aldosterone on the ic Na+ and K+ concentrations of HML was blocked by the aldosterone antagonist canrenone (140 nM). Cortisol at a physiological concentration (40 nM) did not alter ic Na and K concentrations in these cells. The results suggest that aldosterone binding to specific receptors in human mononuclear leukocytes significantly contributes to the regulation of monovalent cation levels in these cells. Thus, this easily accessible model of HML allows study of the effects of mineralocorticoids in relation to their receptors in intact cells in normal and pathological states.
The pancreatic-adrenocortical-pituitary clamp technique for study of counterregulation in humans
Authors: De Feo P. , Perriello G. , Ventura M. M. , Brunetti P. , Santeusanio F. , Gerich J. E. , Bolli G. B.
Published: December 2017

The present experiments were undertaken to develop an approach to analyze the contribution of individual glucose counterregulatory hormones in humans. For this purpose, 24 normal subjects were studied twice: once (control experiments) hypoglycemia was induced by subcutaneous infusion of insulin; and once [pancreatic-adrenocortical-pituitary (PAP) clamp technique] the spontaneous responses of plasma glucagon, growth hormone, and cortisol to hypoglycemia were prevented by intravenous somatostatin and oral metyrapone, respectively, and each hormone was infused at variable rates, which reproduced spontaneous changes in their circulating concentrations in the control experiments. Plasma glucose rate of decrease (0.052 +/- 0.003 vs. 0.06 +/- 0.003 mg X dl-1 X min-1), plasma glucose nadir (49.8 +/- 1.2 vs. 50 +/- 1.0 mg/dl), initial suppression of glucose production (0.22 +/- 0.01 vs. 0.23 +/- 0.01 mg X kg-1 X min-1), subsequent compensatory increase in glucose production (0.54 +/- 0.05 vs. 0.48 +/- 0.04 mg X kg-1 X min-1), and the increase in glucose utilization (0.45 +/- 0.05 vs. 0.42 +/- 0.05 mg X kg-1 X min-1) in PAP clamp and control experiments, respectively, were not significantly different and were significantly correlated. Changes in plasma alanine, lactate, free fatty acids, 3-beta-hydroxybutyrate concentrations were also virtually identical in the PAP clamp experiments and in control experiments. We conclude that the PAP clamp technique can faithfully reproduce the spontaneous hormonal and substrate responses to hypoglycemia and should be useful to assess the contribution of individual hormones during counterregulation by creating an isolated (total or partial) deficiency of a particular hormone without confounding compensatory changes in secretion of other counterregulatory hormones.