bluuebunny/crossref_metadata_2025 huggingface dataset:
Retinyl ester hydrolase and the visual cycle in the chicken eye
Authors: Bustamante J. J. , Ziari S. , Ramirez R. D. , Tsin A. T.
Published: December 2017

The ability of chicken retina and retinal pigment epithelium (RPE) membrane to hydrolyze vitamin A esters ([9,10(-3)H]all-trans- and 11-cis-retinyl palmitate) was studied. Hydrolytic activity within the retina was optimal at acidic pH of 5.0, whereas in the RPE significant hydrolytic activity was exhibited over a broad range of hydrogen ion concentrations. The highest rate of hydrolysis was associated with the all-trans-isomer and located within retina and RPE membranes [the apparent maximal velocity (Vmax) and Michaelis-Menten constant (Km) were 770 pmol.min.-1.mg-1 and 45 microM and 300 pmol.min-1.mg-1 and 3.6 microM, respectively[. Retinyl ester hydrolase activities for 11-cis-retinyl palmitate in the retina and RPE were correspondingly lower (apparent Vmax of 204 pmol.min.-1.mg-1 and Km of 18.5 microM in the retina; apparent Vmax of 131 pmol.min.-1.mg-1 and Km of 4 microM in the RPE). Together with results from other laboratories, results from the present study suggest that chicken retina contains important enzymes to complete the visual cycle.
C1 neurons of neonatal rats: intrinsic beating properties and alpha 2-adrenergic receptors
Authors: Li Y. W. , Bayliss D. A. , Guyenet P. G.
Published: December 2017

Tyrosine-hydroxylase-immunoreactive (TH-ir) neurons (C1 cells) of the rostral ventrolateral medulla (RVL) control sympathetic tone and may be inhibited by the hypotensive drug clonidine. The present study seeks to describe some of the intrinsic properties of bulbospinal C1 cells and to establish whether they have alpha 2-adrenergic inhibitory "autoreceptors." RVL bulbospinal cells (retrogradely labeled with microbeads) were recorded in thin slices from 3- to 7-day-old rats with patch electrodes containing Lucifer yellow. Forty-two of eighty-nine cells were recovered histologically, and 69% were TH-ir. The properties of TH-ir cells could not be distinguished from those of the other bulbospinal cells. Most cells were spontaneously active (0.5-5 spikes/s). Blockade of fast postsynaptic potentials with kynurenic acid plus bicuculline plus strychnine or with a low-Ca(2+)-high-Mg2+ medium failed to reduce their firing rate. The membrane trajectory consisted of one of two patterns interconvertible by injections of depolarizing or hyperpolarizing current: 1) depolarizing ramps leading to spikes or 2) slow oscillations (tetrodotoxin resistant but absent at hyperpolarized potentials) leading to spikes or repolarization. The cells had Ca(2+)-dependent afterhyperpolarizations, large transient outward currents, and a hyperpolarization-activated inward current activated at potentials less than -65 mV, blocked by CsCl, and presumably responsible for a large inward rectification. Of 27 TH-ir cells, 21 had inhibitory responses to alpha 2-adrenergic agonists. These were blocked by idazoxan (n = 9). alpha 2-Agonists activated an inwardly rectifying potassium conductance (KIR). We confirm that bulbospinal C1 cells have intrinsic pacemaker properties and demonstrate that they have alpha 2-adrenergic receptors (autoreceptors) coupled to KIR channels.
Interactions of atrial and brain natriuretic peptides at pathophysiological levels in normal men
Authors: Hunt P. J. , Espiner E. A. , Richards A. M. , Yandle T. G. , Frampton C. , Nicholls M. G.
Published: December 2017

Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) are both circulating plasma hormones that are secreted by the heart and have similar physiological effects. We have shown previously that abrupt increases in plasma BNP in normal humans impair the clearance of ANP from plasma and result in additive physiological effects. Because large increases in plasma ANP are reported to have no effect on plasma BNP levels in patients with heart failure, we have studied ANP-BNP interactions in eight normal male subjects receiving background infusions of BNP (2 pmol.kg-1.min-1 for 5 h). Each subject also received a coinfusion of ANP ("active" day, 2 pmol.kg-1.min-1 for 2 h) or vehicle ("placebo" day) using a balanced random order, single-blind design. Metabolic clearance rate of ANP (mean 4.1 +/- 0.6 l/min) and disappearance rate from the plasma (t1/2 3.4 +/- 0.3 min) were similar to values measured previously in the absence of exogenous BNP. In contrast, steady-state plasma BNP levels were reversibly increased (mean BNP increment 10 pmol/l) during the administration of ANP (P = 0.038). Associated with these changes were significant (additive) physiological effects. Thus the addition of ANP increased plasma and urine guanosine 3',5'-cyclic monophosphate (P < 0.001 for both) and lowered systolic blood pressure (P = 0.049). When ANP was coinfused, significant differences were also observed in urine volume (P = 0.001) and sodium excretion (P = 0.043) between the infusion period (when urine volume and sodium excretion were enhanced) and postinfusion period (when values decreased). Taken together, our findings of similar interactions between ANP-BNP and BNP-ANP infusions occurring at pathophysiological concentrations of these two peptides, suggest that the interactions result from dissociation of prebound hormone, presumably from biological or clearance receptors.
Effects of afferent neural stimulation on critical oxygen delivery: a hemodynamic explanation
Authors: Kirkman E. , Zhang H. , Spapen H. , Little R. A. , Vincent J. L.
Published: December 2017

Injury and activation of somatic afferent nerve fibers may alter critical oxygen delivery (DO2C), the point at which oxygen consumption becomes dependent upon delivery, and hence reduce tolerance to hypovolemia. The present study investigated the mechanism of this. Anesthetized mongrel dogs were divided into two groups: control (n = 6) and those subject to brachial nerve stimulation (BNS; n = 5). Whole body oxygen delivery (DO2I) and consumption were initially similar in both groups. DO2I was reduced by cardiac tamponade to determine DO2C. DO2C was significantly higher in BNS compared with control (11.5: 11.0-16.7 vs. 7.5: 6.9-9.5 ml.min-1.kg-1; median: Q1 - Q3), whereas critical oxygen extraction ratios were lower (54.8: 39.7-61.2 vs. 78.3: 53.5-92.4%). At approximately DO2C, normalized femoral blood flow was lower than renal flow in control (renal-femoral difference 17.4: 8.7-40.0%) but not in BNS (-7.8: -14.8 to +11.8%). These results indicate that activation of somatic afferent nerve fibers elevates DO2C. This could be due to an impairment in peripheral oxygen extraction as a consequence of a redistribution of blood flow away from metabolically active vital organs toward relatively inactive skeletal muscle.
Changes in regional hemodynamics after nitric oxide inhibition during ovine bacteremia
Authors: Lingnau W. , McGuire R. , Dehring D. J. , Traber L. D. , Linares H. A. , Nelson S. H. , Kilbourn R. G. , Traber D. L.
Published: December 2017

We studied the action of nitric oxide synthase (NOS) inhibition on changes in regional blood flow during a continuous infusion of live bacteria. Eighteen ewes were chronically instrumented. After a 7-day recovery period, an infusion of 10(6) colony-forming units/min Pseudomonas aeruginosa was begun. At 24 h, cardiac output increased significantly above baseline in all groups (5.9 +/- 0.4 vs. 8.2 +/- 0.6 l.min 1.m-2), systemic vascular resistance decreased (1,362 +/- 120 vs. 821 +/- 145 dyn.g.cm-5.m-2), and cerebral, cephalic mesenteric, and hindlimb blood flows increased. The animals were then equally and randomly assigned to a bolus of a NOS inhibitor, either 25 mg/kg N omega-nitro-L-arginine methyl ester (L-NAME) or 20 mg/kg N omega-monomethyl-L-arginine (L-NMMA), followed by a continuous infusion of 7 mg.kg-1.min-1 L-NMMA or saline. After NOS inhibition, cardiac index decreased [5.6 +/- 0.1 (L-NAME) and 5.5 +/- 0.4 l.min-1.m-2 (L-NMMA)] and remained significantly decreased for 12 h. 1-NAME decreased carotid and mesenteric blood flows to 64% of the preseptic baseline, and they remained below baseline for 20 h. L-NMMA decreased blood flows only to preseptic baseline values. NOS inhibitors may affect blood flows independently of their hemodynamic effects.
Acute effects of antisense antagonism of a single peptide neurotransmitter in the circadian clock
Authors: Scarbrough K. , Harney J. P. , Rosewell K. L. , Wise P. M.
Published: December 2017

The circadian clock that resides in the suprachiasmatic nucleus (SCN) of the hypothalamus is the major neural pacemaker driving most 24-h rhythms in mammals. Several neurotransmitter peptides are synthesized within this nucleus and communicate rhythmically with other cells in the SCN as well as with cells in other regions of the brain. At the present time, little is known about their role in regulating outputs of the clock. We demonstrate that antisense oligodeoxynucleotides corresponding to the NH2-terminus and the translation start site of vasoactive intestinal peptide (VIP) mRNA infused into the suprachiasmatic region of rats temporarily abolishes the circadian rhythm of corticosterone secretion without influencing stress-related corticosterone secretion in the same animals. Levels of VIP peptide are suppressed 30% on the second day after antisense treatment. These results indicate that a single neuropeptide transmitter in the circadian clock may serve a distinct role in the control of specific circadian rhythms.
Bidirectional effects of hepatic ischemia/reperfusion on E. coli-induced TNF-alpha gene expression
Authors: Epperly N. A. , Lechner A. J. , Johanns C. A. , Webster R. O. , Matuschak G. M.
Published: December 2017

We tested the hypothesis that gram-negative bacteremia (GNB) and brief (30 min) reductions in the hepatic O2 supply by low-flow ischemia differentially modulate tumor necrosis factor-alpha (TNF-alpha) gene expression owing to sequence-specific activation of cyclooxygenase vs. complement (C) pathways. Buffer-perfused Sprague-Dawley rat livers (n = 82) were studied over 180 min after intraportal 10(9) live E. coli serotype 055:B5 (EC) or 0.9% NaCl (NS) at t = 0. Compared with EC and NS controls receiving constant-flow perfusion, sequential GNB and ischemia/reperfusion (I/R) were studied in EC + 30 I/R and NS + 30 I/R livers, in which 30 min of ischemia (I) beginning 0.5 h after EC or NS was followed by 120 min of reperfusion (R). This sequence was reversed in 30 I/R + EC and 30 I/R + NS groups. Bacterial clearance, bioactive and antigenic TNF-alpha, prostaglandin E2 (PGE2), and hepatic O2 uptake and performance were serially assessed. Venous TNF-alpha increased in EC controls to peak at 155 +/- 29 U/ml after 180 min (P < 0.001 vs. NS controls) as did hepatic TNF-alpha mRNA. Both TNF-alpha transcripts and protein levels were markedly attenuated in EC + 30 I/R (P < 0.001 vs. EC) despite equivalent EC clearance by Kupffer cells. Indomethacin (10(-5) M) decreased I/R-induced PGE2 secretion and restored TNF-alpha to control levels. In contrast, TNF-alpha levels in 30 I/R + EC perfusates exceeded those of EC + 30 I/R livers (P < 0.05) and were indistinguishable from EC controls. Allopurinol pretreatment but not heat inactivation of C or infusion of soluble human complement receptor type 1 inhibited TNF-alpha production in 30 I/R + EC organs. These results identify a novel sequence-dependent interaction whereby hepatic O2 deprivation after GNB downregulates TNF-alpha via generation of cyclooxygenase metabolites, whereas ischemia preceding GNB increases cytokine expression via reactive O2 species but not C activation.
Alterations in the cardiovascular control by the chemoreflex and the baroreflex in old rats
Authors: Franchini K. G. , Moreira E. D. , Ida F. , Krieger E. M.
Published: December 2017

The aging process is associated with alterations in the autonomic control of cardiovascular function. In the present study, we observed that reflex bradycardia and tachycardia produced by pressor and depressor agents were attenuated in old (24 mo) rats [70 and 56% of responses in young rats (4 mo), respectively]. In contrast, the bradycardia induced by chemoreflex activation with increasing doses of KCN (60, 100, 140, and 180 micrograms/kg) was greater in old [-30 +/- 19, -155 +/- 32, -198 +/- 15, and -204 +/- 24 beats/ min (bpm)] than in young rats [-13 +/- 6, -75 +/- 20 (P < 0.05), -99 +/- 26 (P < 0.05), and -103 +/- 20 (P < 0.05) bpm]. The chemoreflex-pressor responses tended to be greater in old (12 +/- 6, 47 +/- 6, 54 +/- 7, and 55 +/- 5 mmHg) than in young rats (4 +/- 2, 32 +/- 8, 42 +/- 6, and 44 +/- 4 mmHg), but the differences between both groups were not statistically significant. However, pressor responses were higher in old rats after the chemoreflex-activated bradycardia was attenuated by atropine (4 mg/kg iv) [old (17 +/- 9(1) P < 0.05; 57 +/- 4, P < 0.05; 61 +/- 4; and 64 +/- 5 mmHg) vs. young (3 +/- 2, 29 +/- 9(1) 50 +/- 5, and 58 +/- 7 mmHg)]. Administration of the alpha 1-blocker prazosin (1 mg/kg iv) blunted pressor responses but did not change the bradycardia induced by chemoreflex in both groups. In conclusion, our data indicate that the bradycardia and pressor responses to chemoreflex activation were exaggerated, whereas the heart rate responses (bradycardia and tachycardia) to baroreflex were depressed in old rats.
Homeostatic sleep regulation in habitual short sleepers and long sleepers
Authors: Aeschbach D. , Cajochen C. , Landolt H. , Borbely A. A.
Published: December 2017

Homeostatic sleep regulation in habitual short sleepers (sleep episode < 6 h, n = 9) and long sleepers (> 9 h, n = 7) was investigated by studying their sleep structure and sleep electroencephalogram (EEG) during baseline conditions and after prolonging their habitual waking time by 24 h. In each sleep episode, total sleep time was > 3 h longer in the long sleepers than in the short sleepers. Sleep deprivation decreased sleep latency and rapid eye movement (REM) density in REM sleep more in long sleepers than in short sleepers. The enhancement of EEG slow-wave activity (SWA; spectral power density in the 0.75-4.5 Hz range) in non-REM sleep after sleep loss was larger in long sleepers (47%) than in short sleepers (19%). This difference in the SWA response was predicted by the two-process model of sleep regulation on the basis of the different sleep durations. The results indicate that short sleepers live under a higher “non-REM sleep pressure” than long sleepers. However, the two groups do not differ with respect to the homeostatic sleep regulatory mechanisms.
Swelling-activated K+ transport via two functionally distinct pathways in eel erythrocytes
Authors: Bursell J. D. , Kirk K.
Published: December 2017

Following osmotic swelling, erythrocytes from the European eel, Anguilla anguilla, underwent a regulatory volume decrease. This was prevented by replacement of Na+ with K+ in the suspending medium, consistent with a role for the (normally outward) electrochemical K+ gradient in the volume-regulatory response. The effect of cell swelling on K- transport in these cells was investigated using 86Rb+ as a tracer for K+. Osmotic swelling resulted in an increase in ouabain-insensitive K+ transport that was highest for cells in Cl- and Br- media but which was also significant in I- and NO3- media. Treatment of eel erythrocytes suspended in isotonic Cl- or Br- (but not I- or NO3-) media with the sulfhydryl reagent N-ethylmaleimide (NEM) resulted in a large increase in K+ transport. A quantitative comparison of the pharmacological properties of the “Cl(-)-dependent” NEM-activated pathway with those of the “Cl(-)-independent” pathway mediating swelling-activated K+ transport in cells in Cl(-)-free (NO3- containing) media showed there to be significant differences between them. By contrast, the pharmacological properties of the Cl(-)-independent swelling-activated K+ pathway were indistinguishable from those of the pathway responsible for the swelling-activated transport of taurine, the major organic osmolyte in these cells. A pharmacological analysis of ouabain-insensitive K+ transport in cells swollen in a hypotonic Cl(-)-containing medium showed there to be two components, one with the characteristics of the NEM-activated system, the other showing the characteristics of the Cl(-)-independent swelling-activated pathway. The data are consistent with the presence of two functionally distinct swelling-activated K+ transport mechanisms in eel erythrocytes: a KCl cotransporter that is activated under isotonic conditions by NEM and a Cl(-)-independent, broad-specificity channel that accommodates a diverse range of organic and inorganic solutes.
Central catecholaminergic neurons are involved in expression of the 10-Hz rhythm in SND
Authors: Orer H. S. , Zhong S. , Barman S. M. , Gebber G. L.
Published: December 2017

We studied the effects of adrenoceptor agonists and antagonists on sympathetic nerve discharge (SND) of urethan-anesthetized, baroreceptor-denervated cats. In cats in which a 10-Hz rhythm coexisted with irregular 2- to 6-Hz oscillations in SND, intravenous clonidine, an alpha 2-adrenoceptor agonist, blocked the 10-Hz rhythm without affecting power at lower frequencies. In contrast, power at frequencies < or = 6 Hz was depressed by clonidine in cats in which the 10-Hz rhythm was absent. These effects were reversed by intravenous administration of alpha 2-adrenoceptor antagonists, idazoxan and rauwolscine. Rauwolscine is devoid of affinity for imidazoline receptors. Furthermore, in cats untreated with clonidine, idazoxan and rauwolscine enhanced or induced the 10-Hz rhythm without affecting power at lower frequencies. Prazosin, an alpha 1-adrenoceptor antagonist, selectively blocked the 10-Hz rhythm in SND. Finally, the 10-Hz rhythm in SND was blocked by microinjection of clonidine into the rostral or caudal ventrolateral medulla. The results support the view that central catecholaminergic neurons play a role in expression of the 10-Hz rhythm in SND.
Effect of insulin on regional vascular resistances in conscious rats
Authors: Qadir E. , Porter J. P.
Published: December 2017

In the rat, but not in humans and other mammals, chronic administration of insulin produces hypertension. The present aim was to determine the effect of acute insulin infusion on regional vascular resistances and to determine the neurogenic contribution to the response. Conscious rats were infused with insulin (2 or 6 mU/min) before and after ganglionic blockade with chlorisondamine (5 mg/kg). The low dose of insulin produced an increase in arterial pressure and hindquarter vascular resistance; the high dose produced a gradual decrease in arterial pressure and renal resistance. After ganglionic blockade, the hindquarter vasoconstriction produced by the low dose was abolished. The high dose of insulin produced both hindquarter and renal vasodilation. Thus the low dose of insulin had a selective neurogenic vasoconstrictor effect in rat skeletal muscle vascular beds. With higher doses, direct vasodilatory effects in both skeletal muscle and renal vascular beds appeared. This greater sensitivity of the sympathoexcitatory effects of insulin in rats may explain the ability of chronic insulin infusions to increase blood pressure in this species.
Medullary sympathoexcitatory neurons are inhibited by activation of the medial prefrontal cortex in the rat
Authors: Verberne A. J.
Published: December 2017

Electrical stimulation of the medial prefrontal cortex (MPFC) reduces arterial blood pressure. To investigate the mechanism of this response, the effects of electrical and chemical stimulation of the MPFC on splanchnic and lumbar sympathetic nerve discharge and on the discharges of barosensitive neurons of the rostral ventrolateral medulla (RVLM) were studied in halothane-anesthetized rats. Electrical stimulation (20 Hz, 1 ms, 100 and sympathoinhibitory responses (reduced discharge of the splanchnic sympathetic nerve). Microinjection of glutamate (10 nmol/100 nl) into the MPFC also reduced arterial blood pressure and sympathetic discharge. Electrical stimulation (0.5 Hz, 1-ms pulse pairs, 3-ms interval, 150-300 microA) produced distinct patterns of splanchnic and lumbar sympathetic nerve discharge. A clear sympathoinhibitory phase with an onset latency of 146 +/- 14 ms was observed only in the case of the splanchnic sympathetic nerve activity. Electrical stimulation at depressor sites within the MPFC also inhibited the discharge of 10 of 21 RVLM barosensitive neurons tested. RVLM neurons were never excited by MPFC stimulation. These results indicate that the MPFC contains neurons that form part of a central sympathoinhibitory pathway.
Chronically altered body protein levels following lateral hypothalamic lesions in rats
Authors: Hirvonen M. D. , Keesey R. E.
Published: December 2017

Rats maintaining reduced body weights after lesions of the lateral hypothalamus (LH; LH rats) are characterized by smaller body protein masses. Two experiments were conducted to determine whether this reduced protein mass is actively defended. In the first, it was found that LH rats induced to overeat and restore body weight to the level of nonlesioned controls markedly increased their body fat without significantly increasing body protein. That is, LH rats at normal body weights were notably obese. In the second experiment, body protein losses produced by food restriction in LH rats were both relatively small and proportionally the same as those seen in similarly restricted nonlesioned controls. These observations demonstrate that LH rats retain the capacity for preserving body protein when challenged by either under- or overnutrition. The apparently irreversible reduction in the body protein mass thus appears to be the result of a specific lean tissue downregulation induced by LH damage.
Control of glycolysis in vertebrate skeletal muscle during exercise
Authors: Krause U. , Wegener G.
Published: December 2017

The gastrocnemius muscle of the frog (Rana temporaria) has a high capacity for anaerobic glycolysis from glycogen. Glycolytic metabolites and effectors of phosphofructokinase, particularly the hexose bisphosphates, were followed in muscle during exercise (swimming between 5 s and 5 min), recovery (rest for up to 2 h after 5 min of swimming), and repeated exercise (swimming for up to 60 s after 2 h of recovery). Glycogen phosphorylase and phosphofructokinase were swiftly activated with exercise. The hexose bisphosphates followed markedly different time courses. Fructose 1,6-bisphosphate was transiently increased in both exercise and repeated exercise. This appears to be an effect rather than a cause of phosphofructokinase activation. Glucose 1,6-biphosphate was accumulated only while phosphofructokinase was active and was unchanged at other times. Fructose 2,6-biphosphate showed a 10-fold transient increase on exercise in rested frogs, almost disappeared from the muscle during recovery, and did not change during repeated exercise. Fructose 2,6-biphosphate is a potent activator of phosphofructokinase in vitro under near physiological assay conditions, and it may serve this function also in vivo during exercise. Glucose 1,6-biphosphate could be an activator of phosphofructokinase in repeated exercise when fructose 2,6-biphosphate is not available.
Neuronal pathways involved in abdominal surgery-induced gastric ileus in rats
Authors: Barquist E. , Bonaz B. , Martinez V. , Rivier J. , Zinner M. J. , Tache Y.
Published: December 2017

The 20-min rate of gastric emptying of a noncaloric solution and c-fos expression detected by immunohistochemistry in the brain were monitored 3 h after abdominal surgery performed under 10-min enflurane anesthesia in rats. Abdominal surgery (laparotomy and 1-min manipulation of the cecum) decreased gastric emptying from 60.8 +/- 3.4 to 25.9 +/- 3.4%. Capsaicin applied to the celiac/superior mesenteric ganglia 2 wk before the experiment reduced the delay in gastric emptying induced by abdominal surgery (46.3 +/- 3.4%), whereas perivagal capsaicin application had no effect (23.6 +/- 7.9%). The corticotropin-releasing factor (CRF) antagonist [D-Phe12, Nle21,38,C alpha MeLeu37]CRF-(12--41) injected intracisternally (10-20 micrograms) prevented postoperative gastroparesis induced by surgery, while having no effect on basal gastric emptying. Abdominal surgery increased the number of Fos-positive cells in brain nuclei regulating autonomic outflow: the nucleus of the solitary tract, locus ceruleus, paraventricular nucleus, and supraoptic nucleus of the hypothalamus. These data indicate that capsaicin-sensitive splanchnic afferent fibers and activation of CRF receptors in the brain are part of the neuronal circuitry mediating gastric stasis 3 h after abdominal surgery.
Interleukin-1 beta peptides induce cerebral pial arteriolar dilation in anesthetized newborn pigs
Authors: Shibata M. , Parfenova H. , Zuckerman S. L. , Seyer J. M. , Krueger J. M. , Leffler C. W.
Published: December 2017

Inflammatory cytokines may affect cerebral circulation under pathological conditions. Responses of cerebral pial arterioles to one such cytokine, interleukin (IL)-1 beta and its inhibitor [soluble IL-1 receptor (sIL-1R)] were examined in anesthetized newborn pigs using closed cranial windows. Levels of prostanoids and cyclic nucleotides in periarachnoid cerebral spinal fluid (CSF) were measured. To examine the structure-activity relationship of the parent IL-1 beta molecule, two IL-1 beta fragments with amino acid sequences of 187-204 [IL-1 beta-(187-204)] and 208-240 [IL-1 beta-(208-240)] were tested for their effects on pial arterioles. Diameter changes of pial arterioles were sequentially recorded every 5 min for 30 min after topical application of IL-1 peptides. CSF was sampled at the end of the 30 min. IL-1 beta dose dependently induced arteriolar dilation and increased prostaglandin E2 (PGE2), 6-keto-PGF1 alpha adenosine 3',5'-cyclic monophosphate (cAMP), and guanosine 3',5'-cyclic monophosphate (cGMP). Intravenous indomethacin blocked the IL-1 beta-induced vasodilation, the increased prostanoids, and the increased cAMP, but not the increased cGMP. Neither heat-inactivated IL-1 beta nor IL-1 beta vehicle affected arteriolar diameter or CSF levels of prostanoids. The sIL-1R blocked the IL-1 beta-induced vasodilation and the increased CSF prostanoids. IL-1 beta-(208-240) also induced pial arteriolar dilation; however, its vasodilatory potency was 1,000 times less than that of the whole IL-1 beta molecule. IL-1 beta-(187-204) did not induce pial arteriolar dilation even when its dose was increased to the level of IL-1 beta-(208-240). These results suggest that IL-1 beta, through the activation of membrane-bound IL-1 beta receptors, induces pial arteriolar dilation via mechanisms that involve prostanoids and cyclic nucleotides. The results also indicate that the 208-240 amino acid sequence of IL-1 beta has a sequence-specific physiological function.
Hypoxia-reoxygenation impairs endothelium-dependent relaxation in isolated rat aorta
Authors: Yokoyama S. , Korthuis R. J. , Benoit J. N.
Published: December 2017

The effects of hypoxia followed by reoxygenation on endothelium-dependent relaxation in isolated rat aorta were investigated. Acetylcholine (ACh, 3 nM-10 microM) and calcium ionophore A-23187 (3 nM-300 nM)-induced endothelium-dependent vasorelaxation of isolated rate aortic vessel rings was impaired after 15 min of hypoxia followed by 30 min of reoxygenation. Impairment of ACh-induced relaxation was prevented by pretreatment with the combination of superoxide dismutase (200 U/ml) and catalase (1,000 U/ml). Hypoxia-reoxygenation did not affect sodium nitroprusside (0.1 nM-1 microM)-induced endothelium-independent relaxation nor the dissociation constant of ACh to endothelial M3 muscarinic receptors. Propidium iodide staining of the vascular endothelium revealed a significant increase in the number of dead endothelial cells on the aortic vessel rings following hypoxia-reoxygenation, but not on those pretreated with superoxide dismutase and catalase. These results suggest that hypoxia-reoxygenation impairs endothelium-dependent relaxation of rat aorta by a mechanism that involves oxidant-mediated endothelial cell death.
An endogenous ATP-sensitive glutathione S-conjugate efflux mechanism in Xenopus laevis oocytes
Authors: Ballatori N. , Wang W. , Li L. , Truong A. T.
Published: December 2017

Constitutive efflux mechanisms for reduced glutathione (GSH) and the glutathione S-conjugates S-ethylglutathione (ethyl-SG) and S-(2,4-dinitrophenol)-glutathione (DNP-SG) were examined in Xenopus laevis oocytes. Oocytes were loaded by either microinjection with 50 nl of the 3H-labeled compounds or were exposed to unlabeled 1-chloro-2,4-dinitrobenzene and efflux of DNP-SG synthesized within the oocytes measured spectrophotometrically. Efflux of unlabeled DNP-SG (approximately 1.2 mM intracellular concentration) and microinjected 0.5 mM [3H]DNP-SG was a linear function of time, with approximately 20% released in 3 h at 25 degrees C. [3H] ethyl-SG, 0.5 mM, was released at a comparable rate, whereas only 4% of a tracer dose of [3H]GSH (2.5 mM intracellular GSH) was released in 3 h. Efflux of all three compounds was temperature sensitive and inhibited after ATP depletion but unaffected when Na+ in the culture medium was replaced with K+ or when the pH was changed from 7.5 to either 6.8 or 8.0. Efflux was saturable, with apparent Michaelis constant values of 0.91 +/- 0.19, 0.44 +/- 0.25, and 5.3 +/- 2.2 mM for DNP-SG, ethyl-SG, and GSH, respectively. Bilirubin ditaurate, 0.5 mM, cis-inhibited efflux of 0.5 mM [3H]DNP-SG, 0.5 mM [3H]ethyl-SG, and 2.5 mM [3H]GSH. DNP-SG and ethyl-SG efflux was also cis-inhibited by other glutathione S-conjugates, 0.25 mM 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, 0.5 mM sulfobromophthalein, and 0.5 mM dibromosulfophthalin, but not by 0.25 mM taurocholate. [3H]GSH release (2.5 mM) was unaffected by these compounds or by 10 mM extracellular GSH or methionine. These findings indicate that Xenopus oocytes have an endogenous ATP-sensitive mechanism for extruding glutathione S-conjugates, with properties comparable to ATP-dependent glutathione S-conjugate/organic anion transport systems described in a variety of cell types. However, in contrast to mammalian cells, GSH and ethyl-SG release from Xenopus oocytes was also inactivated after cellular ATP depletion but was not sensitive to membrane depolarization in high-K+ medium or trans-stimulated by extracellular GSH, indicating that efflux of these organic anions from Xenopus laevis oocytes is also mediated by an ATP-sensitive mechanism.
Mechanism of hepatocellular dysfunction during hyperdynamic sepsis
Authors: Wang P. , Chaudry I. H.
Published: December 2017

Because of its central role in metabolism and host defense mechanisms, the liver is thought to be a major organ responsible for the initiation of multiple organ failure during sepsis. It is, therefore, important to discuss whether hepatocellular dysfunction occurs during early sepsis and, if so, whether this occurs prior to hepatocellular damage as evidenced by elevation in serum enzyme levels. Because indocyanine green clearance has been demonstrated to be an early and extremely sensitive measure of active hepatocyte transport function, a technique for repeated measurement of hepatocellular function by in vivo indocyanine green clearance was developed in small animals, such as the rat. Studies have indicated that hepatocellular function is markedly depressed during early stages of polymicrobial sepsis despite the increased cardiac output and hepatic blood flow and decreased peripheral vascular resistance. The depression in hepatocellular function in early, hyperdynamic stages of sepsis does not appear to be due to any reduction in hepatic profusion but is associated with elevated levels of circulating proinflammatory cytokines such as tumor necrosis factor (TNF) and interleukin (IL)-6. Furthermore, administration of recombinant murine TNF-alpha at a dose that does not reduce cardiac output and hepatic perfusion produces hepatocellular dysfunction and increases plasma levels of IL-6. Thus upregulation of TNF and/or IL-6 may be responsible for producing hepatocellular dysfunction during early, hyperdynamic stages of sepsis.
Gonadectomy in the spring reinstates hibernation in male golden-mantled ground squirrels
Authors: Dark J. , Miller D. R. , Zucker I.
Published: December 2017

We tested the hypothesis that continued secretion of gonadal steroids is necessary to suppress hibernation in male golden-mantled ground squirrels in the weeks after the terminal arousal in spring. Juvenile and adult males were gonadectomized or sham gonadectomized 1 wk after the terminal arousal; 64% of castrated and none of the shamcastrated animals resumed hibernation. Latency to resumption of torpor was 9 +/- 2 days from the time of castration, and squirrels underwent 4.3 +/- 0.9 bouts before permanently regaining euthermia. Among squirrels that resumed hibernation, bout duration was significantly shorter and torpor was shallower after castration. Castration as late as 3 wk after the terminal arousal reinstated hibernation. We suggest that the terminal arousal of male squirrels in the spring is provoked by a steroid-independent mechanism similar to that operating earlier in the hibernation season; abandonment of hibernation is contingent on concomitant sustained increases in androgen secretion during the first few weeks of euthermia.
11 beta-hydroxysteroid dehydrogenase modulation of glucocorticoid activities in lymphoid organs
Authors: Hennebold J. D. , Ryu S. Y. , Mu H. H. , Galbraith A. , Daynes R. A.
Published: December 2017

The immunoregulatory effects of glucocorticoids (GCS) are linked to their capacity to alter the production of various species of cytokines associated with immune and inflammatory processes. The present study determined that the influences of GCS within particular lymphoid organs vary, depending on the specific activity of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) within the tissue. This enzyme converts active GCS to inactive metabolites and was found to display greater activity in peripheral than in mucosal lymphoid organs. A direct correlation was found between 11 beta-HSD activity and the preferential production of type 1 cytokines by T-cells residing within certain lymphoid organs. It was established that lymphoid organ 11 beta-HSD was localized in the immobile stromal cell components. Inhibition of 11 beta-HSD activity in vivo reduced type 1 and enhanced type 2 cytokine production by activated T-cells, and it also depressed the ability of animals to generate contact hypersensitivity responses. We conclude that GCS levels can be controlled within lymphoid organs through oxidative inactivation by 11 beta-HSD. GCS action is, therefore, dependent on tissue levels of 11 beta-HSD activity, the number of GCS receptors in a responsive cell, and the concentration of circulating GCS.
Involvement of nitric oxide in inhibitory innervation of urinary bladder of Atlantic cod, Gadus morhua
Authors: Olsson C. , Holmgren S.
Published: December 2017

The aim of this study was to investigate the involvement of nitric oxide (NO) in the nonadrenergic, noncholinergic (NANC) relaxation of the urinary bladder of the Atlantic cod, Gadus morhua. NADPH diaphorase-reactive nerve cells, presumed to be able to produce NO, were found in the vesicular nerve. The cells occurred alone and in ganglia together with stained and unstained cells. The effect of inhibitors of NO synthesis on the relaxation was examined in vitro in isolated muscle preparations. NG-nitro-L-arginine methyl ester (10(-4) M) and NG-nitro-L-arginine (L-NNA; 10(-4) M) decreased the electrically induced relaxation to 32 +/- 6 (n = 8) and 28 +/- 6% (n = 8) of the control, respectively. L-Arginine (10(-3) M) increased the relaxation to 152 +/- 24% (n = 8), without affecting the inhibition by L-NNA. The beta-adrenoceptor antagonist propranolol together with L-arginine analogues abolished the relaxation in 7 of 11 preparations. The NO donor sodium nitroprusside (NaNP) caused a concentration-dependent relaxation of the bladder, with a maximal effect obtained at 10(-4) M. LY-83583 (10(-5) M), a guanylate cyclase inhibitor, decreased both the electrically (n = 8) and the NaNP (10(-6) M, n = 9)-induced relaxation to 69 +/- 5 and 20 +/- 4% of the control, respectively. Together these findings suggest that NO is involved in the NANC regulation of the motility of the urinary bladder of the Atlantic cod.
Blood pressure and fluid-electrolyte balance in mice with reduced or absent ANP
Authors: John S. W. , Veress A. T. , Honrath U. , Chong C. K. , Peng L. , Smithies O. , Sonnenberg H.
Published: December 2017

Atrial natriuretic peptide (ANP)-gene knockout mice of three genotypes (+/+, +/-, and -/-) were maintained on a low-salt diet (0.008% NaCl). They were then fed either the same low-salt diet or a high-salt diet (8% NaCl) for 1 wk. No differences were found among genotypes in daily food and water intakes or in urinary volume and electrolyte excretions. Arterial blood pressures measured in anesthetized animals at the end of the dietary regimen were significantly and similarly increased in -/- compared with +/+ mice on each diet. Renal excretion of fluid and electrolytes was measured in anesthetized mice before and after acute blood volume expansion. No genotype differences were observed before volume expansion. After volume expansion the wild-type (+/+) mice had much greater saluretic responses than either the heterozygous (+/-) or the homozygous mutant (-/-) animals on the low-salt diet but not on the high-salt diet. We conclude that ANP lowers blood pressure in the absence of detected changes in renal function; ANP is not essential for normal salt balance, even on high-salt intake; and ANP is essential for the natriuretic response to acute blood volume expansion on a low-salt but not high-salt intake.
Characterization of cholecystokinin binding sites in goldfish brain and pituitary
Authors: Himick B. A. , Vigna S. R. , Peter R. E.
Published: December 2017

The characterization and distribution of cholecystokinin (CCK)/gastrin binding sites were determined in the goldfish central nervous system (CNS). Binding of 125I-sulfated CCK octapeptide (125I-CCK-8s) in tissue sections was found to be saturable, reversible, time dependent, and displaceable by CCK/gastrin-like peptides. Analysis of saturable equilibrium binding revealed a high-affinity binding site (dissociation constant of 0.706 +/- 0.188 nM), which also displayed high affinity for gastrin-17s and caerulein. Lower affinities were observed for the nonsulfated forms of CCK-8 and gastrin-17. These findings suggest that a single primitive CCK/gastrin receptor exists in the goldfish CNS. The distribution of CCK/gastrin binding sites in the goldfish brain and pituitary revealed high densities within the telencephalon and preoptic hypothalamus, as well as within hypothalamic nuclei associated with the brain feeding center. High densities of binding sites were also localized within the midbrain tegmentum and optic tectum of the midbrain, the facial lobe and vagal lobe of the hindbrain, and within the pituitary pars distalis. Overall, these findings support previous studies that indicate that CCK/gastrin-like peptides play a role in the central regulation of feeding behavior and pituitary hormone secretion in fish.
Fatty acid oxidation modulates the eating response to the fructose analogue 2,5-anhydro-D-mannitol
Authors: Rawson N. E. , Ulrich P. M. , Friedman M. I.
Published: December 2017

The fructose analogue 2,5-anhydro-D-mannitol (2,5-AM) stimulates feeding behavior in rats apparently through its effects on hepatic energy metabolism, where it reduces glucose utilization, traps phosphate, and decreases ATP. The extent to which the magnitude and duration of the eating response are dependent on the ability of the liver to switch to fat oxidation for energy production was investigated by manipulating substrate availability through dietary and pharmacological means. Rats adapted to a high-fat, low-carbohydrate diet preferentially use fat fuels for hepatic energy production and were insensitive to the effects of 2,5-AM on food intake. The lack of an eating response occurred despite similar changes in plasma fuels and liver glycogen compared with rats fed a low-fat, high-carbohydrate diet. In contrast, inhibiting fatty acid oxidation with methyl palmoxirate, which blocks transport of long-chain fatty acids to the mitochondria, potentiated the ability of 2,5-AM to stimulate feeding without altering its effects on plasma and liver fuels. These data demonstrate that the eating response to 2,5-AM is modulated by the availability of fat fuels and implicate a mechanism for initiation of feeding that is not dependent on inhibition of carbohydrate metabolism per se but rather integrates information about the use of both types of fuels.
Effect of vagal and splanchnic nerve section on Fos expression in ferret brain stem after emetic stimuli
Authors: Boissonade F. M. , Davison J. S.
Published: December 2017

Previous studies have demonstrated that intraduodenal hypertonic saline (IHS) induces dense Fos expression within two regions of the ferret dorsal vagal complex (DVC): the area postrema (AP) and the medial subnucleus of the nucleus of the solitary tract (mn). The aims of the present experiments were to determine the peripheral pathways involved in excitation of DVC neurons after IHS and the relative importance of mn and AP excitation in the emetic response to this stimulus. The emetic response and the distribution of Fos were examined after IHS in animals that had received either vagotomy alone, vagotomy and splanchnic nerve section, or sham operation. The emetic response was studied in both awake and anesthetized animals, and Fos induction was studied in anesthetized animals. Vagotomy alone or combined with splanchnic nerve section abolished the emetic response and the area of dense labeling within the mn and reduced but did not abolish the labeling in the AP. It was concluded that both the emetic reflex and the dense expression of Fos within the mn after IHS are dependent on an intact vagus nerve. The excitation of neurons in the AP after IHS is partially dependent on vagal afferents, and the residual labeling that is present in the AP of neurectomized animals may be mediated via a blood-borne route.
Cortisol disposition and production rate in horses during rest and exercise
Authors: Lassourd V. , Gayrard V. , Laroute V. , Alvinerie M. , Benard P. , Courtot D. , Toutain P. L.
Published: December 2017

The influence of a 56-km endurance exercise on cortisol kinetics and production rate was evaluated in six horses administered [3H]cortisol. Exercise resulted in an immediate two- to threefold increase in plasma cortisol, with values returning very rapidly to preexercise levels. During exercise, clearance and steady-state volume of distribution of total cortisol were greatly increased (338 +/- 95 vs. 137 +/- 34 ml.kg-1.h-1 for clearance and 359 +/- 82 vs. 229 +/- 18 ml/kg for volume of distribution), whereas the terminal half-life decreased significantly (0.97 +/- 0.16 vs. 1.55 +/- 0.33 h). The estimated cortisol production rate was 4.41 +/- 1.06 micrograms.kg-1.h-1 at rest and 26.75 +/- 5.11 micrograms.kg-1.h-1 during exercise. We conclude that exercise triggers a large increase (x 6) in the adrenal secretion rate, which is not accurately reflected by the more limited increase (x 2-3) in plasma cortisol concentration, the actual measurement of plasma cortisol clearance being a prerequisite to assessment of adrenal gland function during exercise.
Characteristics of renal sympathetic nerve activity in sodium-retaining disorders
Authors: DiBona G. F. , Sawin L. L. , Jones S. Y.
Published: December 2017

Characteristics of renal sympathetic nerve activity in conscious rats with established congestive heart failure, cirrhosis, or nephrotic syndrome were analyzed using three methods: mean integrated voltage over time, power spectrum analysis, and sympathetic peak detection analysis. Compared with control rats, all three disease models had increased mean integrated voltage. On power spectrum analysis, all three disease models had increased relative power at the heart rate frequency, indicating that it was related to renal sympathetic nerve discharge coupled to the cardiac cycle. Congestive heart failure and nephrotic syndrome rats showed increased relative power in the low-frequency range, whereas cirrhotic and nephrotic syndrome rats showed decreased relative power in the high-frequency range. On sympathetic peak detection analysis, the frequency of sympathetic peaks was greater in the three disease models compared with the control rats. In cirrhotic rats, the distribution of sympathetic peak heights was shifted toward an increased number of peaks of lesser height. It is concluded that basal renal sympathetic nerve activity is chronically increased in these disease models. This is manifest as increased power coupled to the cardiac cycle, which may reflect the disease-specific defects in arterial and cardiac baroreflex control. In cirrhosis, there is possible selective activation of a subgroup of renal sympathetic nerve fibers.
Local cytokine induction by LPS in the rat air pouch and its relationship to the febrile response
Authors: Miller A. J. , Luheshi G. N. , Rothwell N. J. , Hopkins S. J.
Published: December 2017

Peripheral induction of cytokines is a critical event in the induction of febrile responses. The sequence of induction and site of action of these cytokines, however, remain unclear. The objective of the present study was to investigate the kinetics of local and systemic production of interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF), with the aim of identifying the relationship between these cytokines and the febrile response induced by injection of lipopolysaccharide (LPS) into a subcutaneous air pouch in the rat. Intrapouch injection of LPS induced dose-dependent fevers and increases in the concentration of bioactive IL-6 in the plasma. Further studies using 100 microg/kg LPS demonstrated significant increases in local (air pouch) concentrations of bioactive IL-1, TNF and IL-6, and circulating IL-6. No significant increases in TNF or IL-1 were detected in the plasma of the same animals. Local TNF was induced rapidly and peaked 1 h after LPS injection. The kinetics of local IL-1 and IL-6 induction were similar and both peaked after 3 h. The rise in local IL-6 preceded that of plasma IL-6 and reached a peak concentration that was 25-fold higher than that observed in the plasma. The data indicate that IL-1 and TNF act locally at the site of inflammation and that locally induced IL-6 is the important systemic mediator of the response.
Febrile response to tissue inflammation involves both peripheral and brain IL-1 and TNF-alpha in the rat
Authors: Luheshi G. N. , Stefferl A. , Turnbull A. V. , Dascombe M. J. , Brouwer S. , Hopkins S. J. , Rothwell N. J.
Published: December 2017

We investigated the role and interaction between tumor necrosis factor (TNF)-alpha, interleukin (IL)-1, and IL-6 in the development of fever and their involvement in brain and systemic pathways in response to localized tissue inflammation caused by injection of turpentine (TPS) in the rat. Intramuscular injection of 10 microl TPS caused significant increases in body temperature, of up to 2 degrees C, compared with saline-treated animals. Fevers were maximal 7-8 h after injection and were preceded by significant increases in plasma bioactive IL-6. No changes in circulating bioactive IL-1 or TNF-alpha were detected. Systemic injection of IL-1 receptor antagonist (IL-1ra, 2 mg/kg i.p.) or anti-TNF-alpha antiserum (0.4 ml i.v.) almost completely abolished the febrile responses to TPS over 8 h and markedly inhibited the rise in plasma IL-6 bioactivity measured 6 h after TPS. To test the involvement of brain cytokines, anti-TNF-alpha antiserum and IL-1ra were injected intracerebroventricularly. Injections of anti-TNF-alpha antiserum (3 microl/rat i.c.v.) or IL-1ra (400 microg/kg i.c.v.) significantly (P < 0.01 and P < 0.05, respectively) inhibited fever induced by TPS. These data suggest that both localized peripheral and brain IL-1 and TNF-alpha are involved directly in the pyrogenic response to inflammation. The results indicate that, in the periphery, IL-1 and TNF-alpha cause increased production of IL-6, the most likely candidate as a circulating endogenous pyrogen.