Fasting associated with decrease in hypothalamic beta-endorphin.

In rats that were fasted for 2 to 3 days there was a decline in hypothalamic, but not pituitary, beta-endorphin. There was no change in pituitary or hypothalamic adrenocorticotropin content as a result of fasting. Endogenous opiates may be involved in physiological adaptation to fasting.

Activation of renal 1,25-dihydroxyvitamin D3 synthesis by phosphate deprivation: evidence for a role for growth hormone.

In vitro 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] production in kidney slices from normal intact rats averaged 16 +/- 4 pmol/g . h and was increased about 8-fold by phosphate deprivation and 5-fold by calcium deprivation to levels averaging 128 +/- 12 and 84 +/- 19 pmol/g x h, respectively. Hypophysectomy in phosphate-deprived rats completely abolished any increase in 1 alpha-hydroxylase activity, while calcium deprivation in hypophysectomized (hypox) rats resulted in a 4-fold increase in 1 alpha-hydroxylase activity. Replacement of hypox rats fed a low phosphorus diet with pituitary extracts resulted in a 4-fold stimulation of 1 alpha-hydroxylase activity in response to the hypophosphatemic stimulus. However, replacement of hypox rats fed a normal phosphorus diet with pituitary extract stimulated 1 alpha-hydroxylase activity only 2-fold. Replacement of hypox rats fed a low phosphorus diet with GH resulted in a 3.5-fold elevation in plasma 1,25-(OH)2D3 levels, while no such elevation in plasma 1,25-(OH)2D3 levels was observed in similarly treated animals replaced with PRL, ACTH, TSH, or T3. Replacement of hypox rats eating a normal diet with GH resulted in no significant change in plasma 1,25-(OH)2D3 levels. These results suggest that GH is required for maintenance of elevated plasma 1,25-(OH)2D3 levels during dietary phosphate deprivation and that this effect is mediated by increased renal 1,25-(OH)2D3 synthesis.

Vasoactive intestinal polypeptide is synthesized in anterior pituitary tissue.

Previous studies have suggested that vasoactive intestinal polypeptide (VIP) is involved in regulation of PRL secretion within the pituitary gland. In order to determine whether VIP is synthesized in anterior pituitary tissue, we performed three experiments. In all experiments, anterior pituitaries were obtained from male rats. The tissue was then labeled by incubation in leucine-free minimum essential medium containing [3H]leucine, 14 microCi/ml. In Exp I, the labeled tissue was homogenized, centrifuged, and the supernatant was chromatographed on Sephadex G-50F. The fractions indicated a large peak of counts near the void volume and another peak coeluting with VIP. These latter fractions were pooled and subjected to reverse phase HPLC. Fractions from the HPLC indicated: a protein peak, VIP immunoreactivity, and maximum counts immunoprecipitated by anti-VIP serum at the retention time of synthetic porcine VIP. Exp II consisted of perifusion of labeled pituitary quarters over a 120-min period followed by an additional 60 min in the presence of 56 mM KCl. During this latter period of KCl depolarization, a large amount of 3H-labeled material was secreted. These fractions were then chromatographed on Sephadex G-50F, and the fractions coeluting with [125I]porcine VIP were subjected to immunoprecipitation with anti-VIP serum. In addition, all fractions from the Sephadex column were assayed for VIP, and the only activity was at the elution volume of [125I]porcine VIP. In Exp III, the pituitary labeling procedure included 3.6 X 10(-5) M cycloheximide. Subsequently, the tissue was perifused and the perifusate collected during the 60-min 56 mM KCl perifusion phase was pooled and immunoprecipitated with anti-VIP serum. No immunoprecipitable counts were obtained. These experiments indicate that anterior pituitary tissue synthesizes VIP on the basis of the HPLC profile and immunoprecipitation with specific anti-VIP antiserum. These results, in addition to other studies by our laboratory and others, suggest that intrapituitary VIP may be an important regulator of anterior pituitary hormone secretion, particularly PRL.

Plasma free tryptophan, brain serotonin, and an endocrine profile of the genetically obese hyperglycemic mouse at 4--5 months of age.

Genetically obese hyperglycemic mice (ob/ob) were compared with their nonlittermate lean controls at 4-5 months of age with regard to brain serotonin, pituitary ACTH content, and circulating levels of glucose, glucagon, insulin, TSH, T3, T4, total tryptophan and free tryptophan. Brain serotonin pituitary ACTH content, and plasma insulin, glucose, total tryptophan, and free tryptophan were all significantly higher in obese mice than in the controls. TSH, T3, and T4 were not significantly different in obese mice vs. controls, suggesting that the obese mouse is euthyroid. Fasting improved but failed to normalize the glucose and insulin levels or insulin to glucagon ratios. Since serotonin is an important neurotransmitter with regard to hypothalamic-pituitary function and since its levels in the brain are dependent on the availability of tryptophan, the findings of elevated levels of free tryptophan in the plasma and serotonin in the brain of the obese hyperglycemic mouse may help to explain some of the previously observed abnormalities of pituitary hormone secretion in these animals.

A longitudinal hormonal profile of the genetically obese mouse.

Obese mice (C57BL/6J ob/ob) and their lean littermates were studied at various ages from immediately post weaning until 62 weeks of age, at which mortality increased markedly. Several age-related changes were noted. 1) Plasma glucose levels were elevated in obese mice 5-20 weeks and 62 weeks of age, but were similar to those in the lean mice at 20-60 weeks of age. Plasma insulin levels were elevated in obese mice, and there were no age-related differences. 2) Brain serotonin was elevated in obese mice at all ages and increased with age in both obese and lean animals. 3) Pituitary contents of ACTH and beta-endorphin were elevated in young obese mice and increased further as these mice approached their life expectancy. 4) The ratios of ACTH to beta-endorphin immunoreactivities were similar in obese and lean mice, except in obese mice over 50 weeks of age where this ratio was increased. We conclude that: 1) the obese mouse is characterized by hyperinsulinemia and hyperadrenocorticism throughout its life; 2) the insulin resistance of the obese mouse improves at 20 weeks of age, yet deteriorates as its life expectancy is approached; 3) the obese mouse has an elevated brain serotonin content similar to previously described elevations of the putative neurotransmitters dopamine and norepinephrine in these mice; and 4) as the obese mouse approaches its life expectancy, abnormalities may occur in the synthesis, processing, or secretion of ACTH and/or beta-endorphine.

Plasma prolactin-releasing factor-like activity in the amenorrhea-galactorrhea syndrome.

An in vitro bioassay for plasma PRL-releasing factor-like activity has been developed. The method is a three-phase methanol extraction of plasma with extracts of 1.0 ml plasma adjusted to a final volume of 50 microliter. Single 50-microliter aliquots of extract were incubated in 1.0 ml Krebs-Ringer phosphate (KRP) buffer with one rat hemipituitary after a 1-h preincubation. Samples were obtained basally and 30 min after addition of the extract. During each set of incubations, a parallel series of hemipituitaries was incubated in KRP alone. The total nanograms of rat PRL released per mg pituitary tissue during the initial 30 min after preincubation was calculated for all studies. The mean quantity released in KRP alone was considered basal and was subtracted from values obtained during incubation with plasma extracts. The quantity remaining was considered PRL-releasing activity (PRA) of plasma, expressed as nanograms of rat PRL released per mg pituitary. The PRA in plasma from 13 patients with the amenorrhea-galactorrhea syndrome was 132 +/- 17 ng/mg pituitary (X +/- SE), which was significantly greater (P less than 0.001) than the PRA in plasma from eight matched controls [31 +/- 10 ng/mg pituitary (X +/- SE)]. The patients' individual PRL levels were elevated (range, 48-248 ng/ml), and when compared to the PRA in the samples, a highly significant (P less than 0.001) positive correlation evolved. These results indicate that a circulating PRL-releasing factor-like material present in normal plasma is higher in plasma from hyperprolactinemic patients in direct relationship to the PRL concentration. It is possible that this material is related to the pathogenesis of PRL-secreting pituitary disorders.

The interrelationships among prolactin, 1,25-dihydroxyvitamin D, and parathyroid hormone in humans.

Serum PRL, parathyroid hormone (PTH), and plasma 1,25-dihydroxyvitamin D [1,25(OH)2D]concentrations were measured in 6 women and 2 men with hyperprolactinemia, 6 normal men and 7 normal women, 4 men and 4 women with primary hyperparathyroidism, and 16 men and 4 women with Ca nephrolithiasis. Plasma 1,25(OH)2D and serum parathyroid hormone (PTH) concentrations were normal in the women and men with hyperprolactinemia. In patients with primary hyperparathyroidism and elevated serum PTH, plasma 1,25(OH)2D concentrations were elevated but serum PRL levels were normal. Likewise, serum PRL levels were normal in patients with Ca nephrolithiasis who had significantly elevated plasma, 1,25(OH)2D concentrations and normal serum PTH concentrations. Thus, hyperprolactinemia due to pituitary adenoma or idiopathic hypersecretion is not accompanied but elevated plasma concentrations of 1,25(OH)2D.

Hormonal, metabolic and morphologic studies of aged C57BL/6J obese mice.

Genetically obese mice (C57BL/6J-ob/ob), fed ad libitum, demonstrated a precipitous increase in the spontaneous death rate after 50 weeks. The first signs of morbidity were a ruffled hair coat and a progressive motor ataxia. Necropsy revealed that obese mice had pale and fatty livers, urolithiasis and grossly distended bladders. Microscopically, the hepatocellular changes observed in all aged obese mice included: a loss of orientation of hepatocytes, an enormous variability in the size of both hepatocytes and their nuclei, and an extensive deposition of both large and small lipid droplets, confirmed by an increase content of triacylglycerols. A subacute-to-chronic, multifocal, necrotizing hepatitis was also present. Kidneys from aged obese mice contained hypertrophied glomeruli and increased PAS-stained material. Tubular dilation with compaction of the tubular cells was also seen. There were no significant alterations in the microanatomy or mineralization of femurs from obese mice, yet there was a significant increase in plasma alkaline phosphatase activity. In obese mice at 62-63 weeks of age, hyperglycemia was present even in spite of hyperinsulinemia. Pituitary immunoreactive ACTH and its molar ratio to pituitary immunoreactive beta-endorphin were also increased in obese mice at this age. Even though the etiology of the decreased lifespan of genetically obese mice remains uncertain, the possibility is discussed that an overall defect in the central nervous system may be involved.

Prolactin and growth hormone responses in the amenorrhea-galactorrhea syndrome.

Fifteen patients with hyperprolactinemia, amenorrhea, and galactorrhea were studied; 10 of these patients had apparent tumors. Nine of 12 patients had a suppression of prolactin with oral L-dopa, and all of 4 patients undergoing dopamine infusion had suppression of prolactin. Eleven and 8 patients underwent chlorpromazine stimulation and insulin-induced hypoglycemia, respectively; none responded. Three of the 4 patients did not show a growth hormone response with dopamine infusion, and L-dopa testing failed to achieve a growth hormone response in 7 of 10 patients. The data suggest an intact dopaminergic inhibition of prolactin with exogenous agents. However, inhibition of endogenous dopamine by chlorpromazine failed to elicit a prolactin response. The disordered growth hormone response to dopaminergic agents suggests a central disorder of dopamine generation. The possible implication of these results with respect to the pathogenesis of hyperprolactinemia is discussed.

Influence of endogenous opioids on the response of selected hormones to exercise in humans.

To examine the influence of endogenous opioids on the hormonal response to isotonic exercise, eight males were studied 2 h after oral administration of placebo or 50 mg naltrexone, a long-lasting opioid antagonist. Venous blood samples were obtained before, during, and after 30 min of bicycle exercise at 70% VO2max. Naltrexone had no effect on resting cardiovascular, endocrine, or serum variables. During exercise epinephrine was higher [mean 433 +/- 100 (SE) pg/ml] at 30 min with naltrexone than during placebo (207 +/- 26 pg/ml, P less than 0.05). Plasma norepinephrine showed the same trend but the difference (2,012 +/- 340 pg/ml with naltrexone and 1,562 +/- 241 pg/ml with placebo) was not significant. Plasma glucose was higher at all times with naltrexone. However, the difference was significant only 10 min into recovery from exercise (104.7 +/- 4.7 vs. 94.5 +/- 2.8 mg/dl). Plasma growth hormone and cortisol increased during recovery and these elevations were significantly (P less than 0.05) augmented by naltrexone. Plasma vasopressin and prolactin increased with exercise as did heart rate, blood pressure, lactic acid, and several serum components; these increases were not affected by naltrexone. Psychological tension or anxiety was lower after exercise compared with before and this improved psychological state was not influenced by the naltrexone treatment. These data suggest that exercise-induced activation of the endogenous opioid system may serve to regulate the secretion of several important hormones (i.e., epinephrine) during and after exercise.

Peripheral motilin administration stimulates feeding in fasted rats.

Although the physiologic function of the gastrointestinal hormone motilin remains uncertain, plasma levels of this peptide vary with migrating myoelectric complexes (MMCs) in the small intestine. In the fed state, both MMCs and plasma motilin are suppressed. During fasting, cyclical peaks of motilin in plasma occur at the same time as Phase III of the MMC cycle occurs in the duodenum. This dependence of motilin concentrations in plasma on the feeding state of the animal prompted an investigation of the effects of motilin on feeding behavior. Intraperitoneal injection of motilin into fasted, but not fed, rats stimulated eating in a dose dependent manner. A significant stimulation of feeding was seen at doses of 5 and 10 micrograms/kg. Sated rats did not eat whether injected with motilin or vehicle. The feeding response to motilin was blocked by prior injection of the rats with naloxone, naltrexone, or pentagastrin. The dose response suppression of food intake by naloxone was similar in fasted animals treated with motilin or vehicle. Motilin may function as a hunger hormone during periods of fasting.

Central administration of motilin stimulates feeding in rats.

Peripheral administration of motilin has been found to stimulate feeding behavior in rats. Since motilin immunoreactivity has been found in discrete brain sites, we tested the effect of motilin administered intracerebroventricularly on feeding in rats. Injection of 1 microgram of motilin significantly increased food consumption at 2 hours, 22 hours, and at 24 hours in animals tested either at or 2 hr prior to lights out. Motilin also significantly increased food consumption in animals maintained under continuous lights-on at 2 hours (488% of control), 22 hours (128% of control), and at 24 hours (140% of control).

SU-E-J-46: Use of Brainlab-ExacTrac IGRT with Dot-Decimal Electron Compensator(BolusECT) for Accurate Patient Positioning.

PURPOSE: To Validate Brainlab-ExacTrac IGRT with Dot-Decimal electron compensator(BolusECT) for accurate patient positioning. METHODS: Dot-Decimal electron compensators(BolusECT) are used to achieve conformal dose distributions and greater sparing to normal structures for cases with varying patient contours and treated with single electron beam.Setup for such patients is difficult to verify due to the small clearance between theelectron-cone and bolus. Hence additional verification is needed for fast setup to avoid discomfort for the patient. In this study we have explored theuse of Brainlab-ExacTrac IGRT system for such setups.A Preplan isrequired to construct the BolusECT and a post-plan is used for verifying the treatment plan. Before the patient is scanned again with BolusECT for post-plan, tungsten BBs were drilled in the Bolus at different depths and the holeswere filled with wax. Tungsten-BBs have high density and avoid high cost ofgold seeds.A wax phantom and BolusECT were scanned with BBs and a plan was generated. The plan was send to ExacTrac for DRR-generation.The Wax phantom with BolusECT was then set slightly off centered on thetreatment couch and the ExactTrac system was used to align the Phantom. The ExacTrac system was also tried on two patients for accurate setup. RESULTS: The ExactTrac system correctly identified the tungsten-BBs and aligned the phantom within +1mm. The SSD was then verified and checked with the planned SSD.The SSD was found to be within +0.5mm. In both patient setups the ExacTrac system gave Lateral, Longitudinal and Vertical Shifts within +1mm and the SSD was found to be within +0.3mm of the planned SSD. CONCLUSIONS: The BrainLab- ExacTrac IGRT system isan excellent,fast way of aligning the BolusECT. This avoids discomfort for patients by preventing longer setup times. Use of tungsten-BBs lowers the cost compared to gold seeds. Phantom verification should be performed before implementing this system for actual patient use. The SSDs must be verified after setup.

Evidence that serotonin stimulates a prolactin-releasing factor in the rat.

Methanol extracts of rat plasma resulted in release of prolactin (PRL) from rat hemipituitaries in vitro with a linear log-dose relationship. This prolactin-releasing factor (PRF)-like activity was not altered in plasma from rats treated with bromocryptine or chlorpromazine despite significant suppression and stimulation of plasma PRL levels, respectively. Fluoxetine, a serotonin reuptake inhibitor, plus 5-hydroxytryptophan, the immediate precursor of serotonin, markedly stimulated both plasma PRL and plasma PRF-like activity. Neither fluoxetine, 5-hydroxytryptophan, nor the combination directly stimulated PRL release from rat pituitary tissue in vitro. We conclude that serotonergic stimulation augments PRL release via a PRF.

Erythrocyte aldehyde dehydrogenase and disulfiram-like side effects of hypoglycemics and antianginals.

Disulfiram-like responses to various drug therapies are caused by elevated ethnol-derived blood acetaldehyde concentrations resulting from drug-induced inhibition of aldehyde dehydrogenase enzymes. We have found that the nitrate ester antianginal drugs, isosorbide dinitrate and nitroglycerin, are potent inhibitors of human erythrocyte aldehyde dehydrogenase. To further characterize this drug-induced enzyme inhibition, erythrocyte aldehyde dehydrogenase activities were measured in patients undergoing therapy with nitrate ester antianginals (isosorbide dinitrate and nitroglycerin) and sulfonylurea hypoglycemics (chlorpropamide and tolazamide). The erythrocyte enzyme was decreased by approximately 25% in sulfonylurea-treated patients, whereas in the nitrate ester-treated patients, an 88% inhibition was observed. The minimal enzyme inhibition in the sulfonylurea-treated group was unexpected because these therapies have well-documented disulfiram-like side effects. This weak inhibition contrasted with the severe inhibition observed in the nitrate ester-treated group where the disulfiram-like side effects are not considered a serious clinical problem. This apparent anomaly was attributed to differences in inhibition of the erythrocyte and liver aldehyde dehydrogenase by the parent drugs and their hepatic metabolites.

Plasma adrenocorticotropin and cortisol responses to submaximal and exhaustive exercise.

Adrenocorticotropic hormone (ACTH) levels were compared before and after submaximal and exhaustive isotonic exercise in six normally active college students (3 men and 3 women). Each subject participated in three experiments conducted at the same morning hour. Venous plasma was obtained before and immediately after 20-min runs at 65 and 80% of maximal O2 consumption (VO2 max) and after a run of gradually increasing work intensity which resulted in exhaustion (100% VO2 max) in 12.6 +/- 1.3 min. ACTH (mean +/- SE) was 48 +/- 15, 57 +/- 12, and 61 +/- 11 pg/ml before the 65, 80, and 100% VO2 max runs, and increased to 61 +/- 15 (NS), 128 +/- 18 (P less than 0.05), and 292 +/- 72 (P less than 0.05) pg/ml, respectively. Plasma glucose, growth hormone, cortisol, and lactic acid concentrations increased in a similar fashion. Cortisol and ACTH levels were significantly correlated at the higher levels of exertion: r = 0.18 (NS) for the 65% VO2 max run, r = 0.65 (P less than 0.05) for the 80% VO2 max run, and r = 0.64 (P less than 0.05) for the run to exhaustion. Both the change in ACTH with exercise and its postrun concentration were significantly related to the change in plasma lactic acid (r = 0.65, P less than 0.05) and the postrun plasma lactic acid (r = 0.64, P less than 0.05). We conclude that exercise-induced increases in plasma ACTH and their correlation with circulating cortisol depend on the intensity of isotonic exercise. Our observations also suggest that plasma lactic acid may influence ACTH release during exercise.

Age-related changes in central nervous system beta-endorphin and ACTH.

Aging is associated with alterations in mood, thermoregulation, pain threshold, and stress response. Because these functions may be modulated by endogenous opiates, we measured immunoreactive ACTH with beta-endorphin in discrete brain areas and pituitary glands from rats aged 6 weeks (young), 6 months (mature), and 20-24 months (senscent). Beta-Endorphin and ACTH declined significantly with aging in the hypothalamus and corpus striatum. Beta-Endorphin and ACTH increased in the frontal lobe during early life; however, no change was noted after maturity. A discordant response with age was noted in the pituitary in that (ACTH did not change, while beta-endorphin increased early in life without change after maturity. Cerebellar tissue exhibited no immunoreactive ACTH or beta-endorphin. Age-related changes in brain and pituitary beta-endorphin and ACTH must be considered in the evaluation of the physiological aging process and when comparing studies of these neuropeptides.

Treatment of thrombotic thrombocytopenic purpura by exchange transfusion.

Three successive patients with thrombotic thrombocytopenic purpura (TTP) were treated by exchange transfusion on a total of 5 occasions in addition to receiving more conventional therapy such as corticosteroids, platelet inhibitors, heparin, and splenectomy. Dramatic relief of symptoms and objective improvement in hematologic values occurred within 24 hours of each exchange. One patient subsequently died of complications resulting from a massive hemorrhage, but in hematologic remission. The remaining 2 are now apparently well. These observations appear to confirm recent reports of a beneficial effect of exchange transfusion in TTP. Although the reason for its effectiveness is not yet known, exchange transfusion may be a highly useful adjunct in the treatment of this disorder and further evaluation of this form of therapy appears warranted.

Cardiovascular fitness and selected adrenal hormone responses to cognitive stress.

Individuals manifesting high maximal oxygen consumption (VO2max) have augmented adrenal responsivity to physical stress. We subjected 17 males representing a wide range of VO2max to cognitive stress and measured their forearm venous plasma epinephrine, norepinephrine, and cortisol responses. Six repeated blood samples were collected at resting, stress, and recovery intervals. Significant increases were observed for epinephrine (X +/- SD; rest, 50 +/- 30; stress, 81 +/- 77 pg/ml; p less than or equal to 0.05) and norepinephrine (rest, 404 +/- 235; stress, 481 +/- 314 pg/ml; p less than or equal to 0.05). Cortisol changes were not significant. A positive association (r = 0.54; p less than or equal to 0.05) was observed between epinephrine and VO2max during the first collection of the stress period. An inverse association (r = 0.49; p less than or equal to 0.05) was present between VO2max and norepinephrine. No significant correlations occurred with cortisol. These results indicate that enhanced cardiovascular fitness is characterized by augmented epinephrine responsivity to cognitive stress. This association exists in the presence of an inverse relationship between plasma free norepinephrine response and VO2max.

Antisera to vasoactive intestinal polypeptide inhibit basal prolactin release from dispersed anterior pituitary cells.

Vasoactive intestinal polypeptide (VIP) has been identified in hypothalamic tissue, is secreted into hypophysial portal blood, and stimulates prolactin (PRL) release in vivo and in vitro. It has been proposed, therefore, that VIP is a physiologic PRL-releasing factor. In this study, we confirm that VIP stimulates PRL release from rat pituitary cells in vitro, and demonstrate that an anti-VIP antiserum blocks VIP-induced PRL secretion. Surprisingly, the anti-VIP antiserum inhibited basal PRL secretion from rat pituitary cells in 3 separate experiments. Data from these experiments were pooled, as the responses were similar, revealing basal PRL release of 10.7 +/- 1.3 ng rPRL/10(5) cells (X +/- SE), while anti-VIP antisera significantly inhibited release to 4.4 +/- 0.6 ng rPRL/10(5) cells (p less than 0.001). PRL release in incubates containing control non-immune sera did not differ from basal release, 8.1 ng rPRL/10(5) cells. A further control experiment was conducted wherein cells were incubated with an anti-ACTH antiserum, representing another hyperimmune serum, which had no effect on PRL secretion. These data suggest that VIP, in addition to its possible role as a hypothalamic-derived PRL-releasing factor, may play a role within the pituitary as a regulator of basal PRL secretion.