bluuebunny/crossref_metadata_2025 huggingface dataset:
A molecular genetic method for genotyping fatty (fa/fa) rats
Authors: Smoller J. W. , Truett G. E. , Hirsch J. , Leibel R. L.
Published: December 2017

After three decades of physiological research, the precise nature of the genetic lesion in Zucker fatty (fa/fa) rats remains unknown. Several methods have been used to identify preobese rats to detect the earliest phenotypic effects of the fa mutation. Most of these methods have used phenotypic characteristics that are not reliable until the second week of life, when increased adiposity is already evident. We have used a restriction fragment length polymorphism (RFLP) for a human genomic DNA probe (VC85) that is tightly linked to the fa locus on rat chromosome 5 to genotype the F2 progeny of a Zucker (13M) x Brown Norway (BN) fa/+ F1 intercross. Sixty-four rats, comprising five litters, were killed at 5-6 wk of age. DNA was isolated either from tail at age 4-7 days (36 rats) or from organs at the time of death (28 rats). Adiposity was scored using inguinal fat pad weight as a percentage of body weight. RFLP analysis was > 99% accurate in identifying obese (fa/fa) rats. This molecular genetic method can be used to genotype fatty rats from an appropriate genetic cross at any age, even prenatally. Moreover, this method can distinguish heterozygous from homozygous littermates, enabling an analysis of gene dosage effects.
Cardiac-renal-neural reflex plays a major role in natriuresis induced by left atrial distension
Authors: Miki K. , Hayashida Y. , Shiraki K.
Published: December 2017

The role of cardiac-renal-neural reflex in the natriuresis induced by left atrial balloon inflation was investigated in conscious dogs. Female mongrel dogs were assigned randomly to 1) sham-operated (n = 8), 2) cardiac-denervated (n = 6), and 3) renal-denervated (n = 8) groups. The dogs were chronically instrumented with a bipolar stainless steel wire electrode for measurement of renal sympathetic nerve activity (RSNA). Balloon inflation induced a step increase in left atrial pressure (Pla) by 7.7 +/- 1.7 mmHg, a step decrease in RSNA (-66.6 +/- 5.5%), and concomitant increases in urine flow (441 +/- 142%), osmolal excretion (60 +/- 12%), and sodium excretion (300 +/- 69%) in sham-operated dogs. Renal denervation abolished the diuresis and natriuresis during balloon inflation. Chronic cardiac denervation abolished also the diuresis and natriuresis in the face of a similar increase in Pla. RSNA did not change significantly throughout the experimental period in cardiac-denervated dogs. It is concluded that a sustained reduction of RSNA originating from left atrial mechanoreceptors plays a major role in the natriuresis during left atrial distension in conscious dogs.
Cardiovascular actions of ANF: contributions of renal, neurohumoral, and hemodynamic factors in sheep
Authors: Charles C. J. , Espiner E. A. , Richards A. M.
Published: December 2017

Whereas the short-term effects of atrial natriuretic factor (ANF) are well documented, less is known concerning the possible hypotensive action of sustained low-dose ANF infusions. Accordingly, we have examined the sequential renal, hormonal, and hemodynamic responses to 48-h low-dose ANF infusions in eight normotensive conscious sheep and evaluated the effects of these infusions on autonomic (baroreceptor), adrenocortical, and pressor responsiveness to exogenous stimulation. Plasma ANF levels tended to rise in response to ANF infusions, but the difference between ANF and control day levels was not significantly different. Systolic arterial pressure (SAP) was significantly lower after ANF infusion (P = 0.01) and was associated with a reduction in calculated total peripheral resistance (CTPR, P = 0.007). Mean arterial pressure also tended to be lower (P = 0.08) in response to ANF. No change was seen in urinary volume or sodium excretion or in plasma angiotensin II (ANG II), aldosterone, or cortisol levels. ANF significantly attenuated the pressor response to exogenous ANG II (P = 0.01) but did not affect the adrenocortical responsiveness or autonomic (baroreceptor) responsiveness to exogenous stimulation. This study demonstrates that chronic low-dose infusions of ANF that barely elevate plasma ANF levels induce significant cardiovascular effects, including lowering of SAP associated with a fall in CTPR and attenuation of pressor responsiveness to physiological increments in ANG II, thus providing further support for the importance of ANF in blood pressure homeostasis in normal sheep.
Developmental changes in renin gene expression in ovine kidney cortex
Authors: Carbone G. M. , Sheikh A. U. , Rogers S. , Brewer G. , Rose J. C.
Published: December 2017

The ontogeny of renin mRNA and renin content from renal cortical slices was studied in two groups of ovine fetuses at 92-94 days (0.64 gestation) and at 138-142 days (0.96 gestation), newborn lambs (0.4-2 days old), and adult sheep. Renal renin mRNA was identified by hybridization with a 32P-labeled full length rat renin cDNA. Renal renin content was measured as nanograms of angiotensin I generated per hour (active renin). There was a significant age effect on renin mRNA levels (F = 10.0, P < 0.001); values increase significantly between 0.64 and 0.95 g (P < 0.005), remain elevated in the newborns (P < 0.05), and subsequently decline in adulthood (P < 0.005). Likewise, renal renin content was significantly higher in late gestation fetuses and newborn lambs than in early gestation and adults (F = 8.3, P < 0.003). The renal renin content was strongly correlated with renin mRNA levels (R = 0.88, P < 0.0001). These results suggest that 1) the renin gene is developmentally regulated in the ovine kidney and 2) the renal content of active renin in basal conditions is regulated, at least in part, by events at the transcriptional level.
Mechanisms of renal vasodilation after protein feeding: role of the renin-angiotensin system
Authors: Woods L. L.
Published: December 2017

These studies were designed to determine the importance of the renin-angiotensin system (RAS) in the renal hemodynamic response to acute protein feeding. In chronically instrumented conscious dogs on a normal (80 meq/day) sodium intake, a 10 g/kg meal of raw beef caused glomerular filtration rate (GFR) to increase from 68 +/- 6 to 86 +/- 6 ml/min and effective renal plasma flow (ERPF) to increase from 211 +/- 14 to 263 +/- 15 ml/min. Plasma renin activity (PRA) was 0.44 +/- 0.14 ng ANG I.ml-1 x h-1 and did not change significantly. When the protocol was repeated during infusion of captopril, GFR increased from 67 +/- 11 to 97 +/- 10 ml/min, and ERPF rose from 264 +/- 74 to 392 +/- 82 ml/min after the meat meal. The dogs were then placed on a low-salt diet (approximately 7 meq/day) to physiologically activate the RAS. In sodium-restricted dogs, GFR increased from 71 +/- 7 to 104 +/- 10 ml/min and ERPF increased from 226 +/- 15 to 299 +/- 21 ml/min after the meat meal. PRA was 3.1 +/- 1.0 ng ANG I.ml-1 x h-1 and did not change. Thus neither blockade of the RAS with captopril nor activation of the RAS by salt restriction reduced the renal hemodynamic response to a meat meal. These data indicate that the RAS is relatively unimportant in the renal hemodynamic response to acute protein feeding.
Melatonin injections affect circadian behavior and SCN neurophysiology in Djungarian hamsters
Authors: Margraf R. R. , Lynch G. R.
Published: December 2017

We investigated the effects of daily melatonin (MEL) injection on phase angle of entrainment, duration of wheel-running activity (alpha), and frequency of suprachiasmatic nuclei (SCN) neuronal discharge in the photo-nonresponsive phenotype of the Djungarian hamster, Phodopus sungorus. Photo-nonresponsiveness is characterized by an absence of physiological adjustments to short days (SD). With respect to wheel-running activity, photo-nonresponsive hamsters have a large negative phase angle of entrainment and a compressed alpha under SD. These hamsters also have a delayed nocturnal MEL pulse. These circadian differences are correlated with the daily profile of SCN neuronal activity. In the present experiments, daily MEL injections to photo-nonresponsive hamsters resulted in molt, gonadal regression, and expansion in alpha until entrainment to lights off. Vehicle-injected controls did not exhibit any of these responses. SCN neuronal activity patterns recorded from MEL-injected photo-nonresponders, but not vehicle-injected controls, resembled electrical activity profiles of photoresponsive hamsters. These results demonstrate that MEL induces "photoresponsiveness" in previously photo-nonresponsive hamsters, that MEL modifies circadian behavior to resemble that of photoresponders, and that MEL injections affect the circadian rhythm of SCN neuronal firing.
Chemically induced hypotension increases PGE2 release and depresses macrophage antigen presentation
Authors: Ertel W. , Singh G. , Morrison M. H. , Ayala A. , Chaudry I. H.
Published: December 2017

Hemorrhagic shock causes severe depression of macrophage functions and is associated with increased susceptibility to sepsis. Because hemorrhagic shock and resuscitation encompasses several pathophysiological conditions, such as hypotension, low-flow conditions, hypoxia, and reperfusion injury, it remains unknown whether severe hypotension in the absence of blood loss has any adverse effects on macrophage functions. To study this, systemic arterial hypotension was induced in C3H/HeN mice for 15 min by intravenous infusion of sodium nitroprusside or ATP-MgCl2. Peritoneal macrophages (PM) was harvested 20 h later with lavage. Antigen presentation was measured by coculturing PM with the D10.G4.1 Th cell clone. Tumor necrosis factor (TNF), interleukin (IL)-6, IL-1, and prostaglandin (PG) E2 levels in supernatants of PM stimulated with lipopolysaccharide were measured with bioassays or radioimmunoassay. Systemic arterial hypotension resulted in a significant decrease of PM capacity to present antigen. Although the release of TNF, IL-6, and IL-1 by PM was unaltered after hypotension, PGE2 release by PM was significantly elevated compared with the control group. These data indicate that chemically induced systemic arterial hypotension without blood loss leads to a depression of antigen presentation, which may be caused by elevated release of the immunosuppressive eicosanoid PGE2.
Renal and systemic adaptation to sodium restriction in aging rats
Authors: Jover B. , Dupont M. , Geelen G. , Wahba W. , Mimran A. , Corman B.
Published: December 2017

The influence of age on the systemic and renal adaptation to dietary sodium restriction was assessed in 10-, 20-, and 30-mo-old female WAG/Rij rats. In control conditions, mean arterial pressure (MAP) was similar in all rats and plasma renin activity (PRA) was lower in 30- than in 10- and 20-mo-old rats (2.5 +/- 0.6, 5.1 +/- 0.4, and 3.9 +/- 1.0 ng ANG I.ml-1.h-1, respectively). Dietary sodium restriction was associated with a reduction in MAP in 30-mo-old rats, whereas no change occurred in 10- and 20-mo-old rats. Impairment in the early (days 1-6) renal adaptation to salt restriction was observed in 30- compared with 10- and 20-mo-old rats (6-day cumulative sodium excretion of 728 +/- 139, 437 +/- 53, and 478 +/- 37 mumol, respectively). During the 7- to 12-day period, MAP stabilized in the oldest rats and cumulative sodium excretion became similar to that of other age groups. The early increase in PRA and urinary aldosterone excretion observed in 10- and 20-mo-old rats was consistently blunted in 30-mo-old rats. These findings suggest that the delayed response of the renin-angiotensin-aldosterone system has a major role in the impaired renal and systemic adaptation to dietary sodium removal in senescent rats.
Abdominal vagotomy decreases the satiating potency of CCK-8 in sham and real feeding
Authors: Joyner K. , Smith G. P. , Gibbs J.
Published: December 2017

To compare the effect of total abdominal vagotomy on the satiating potency of cholecystokinin COOH-terminal octapeptide (CCK-8) in sham feeding and real feeding, we administered CCK-8 intraperitoneally (2, 4, and 8 micrograms/kg) 1 min before a sham feeding or real feeding intake test. CCK-8 was at least as potent for inhibiting intake during sham feeding as it was during real feeding. Total abdominal vagotomy decreased the satiating potency of all three doses of CCK-8 equally in both feeding conditions. These results are consistent with the proposal that abdominal vagal afferent fibers are as necessary for the mediation of the satiating effect of CCK-8 in sham feeding as they are in real feeding.
Angiotensin type 2 receptors mediate depressor phase of biphasic pressure response to angiotensin
Authors: Scheuer D. A. , Perrone M. H.
Published: December 2017

Angiotensin (ANG) can produce a biphasic arterial pressure response, i.e., an increase followed by a decrease. Because ANG type 1 (AT1) receptors mediate the pressor response to ANG, we hypothesized that the opposing depressor action is mediated by the ANG type 2 (AT2) receptors. In thiobutabarbital (Inactin)-anesthetized rats bolus injections of angiotensin III (ANG III; 100, 300, and 1,000 ng/kg iv) produced peak increases in MAP at 20 s of 13.4 +/- 1.4, 20.1 +/- 2, and 27.5 +/- 2.8 mmHg and maximum decreases in pressure at 120 s of -6.3 +/- 1.5, -6.8 +/- 2.2, and -11.4 +/- 4.9 mmHg. During blockade of the AT1 receptors with DuP 753 (losartan, 10 mg/kg) the increases in MAP were eliminated (P < 0.01), whereas the depressor responses (-24.7 +/- 8, -32.8 +/- 9.3, and -42.0 +/- 10.0 mmHg) were significantly (P < 0.05) larger. In separate groups of rats, combined blockade of both AT1 and AT2 receptors eliminated all changes in MAP in response to ANG III, whereas blockade of AT2 receptors alone enhanced the pressor response to ANG III. During AT1 receptor blockade angiotensin II also caused consistent decreases in pressure, which were inhibited during combined blockade of AT1 and AT2 receptors. Therefore, we have demonstrated that the AT2 receptors mediate a depressor response to ANG.
Large enhancement of canine taste responses to amino acids by salts
Authors: Ugawa T. , Kurihara K.
Published: December 2017

The effects of salts on the canine taste responses to amino acids were examined by recording the activity of the chorda tympani nerve. 1) The tonic responses to most amino acids examined were significantly enhanced by the presence of various salts. 2) The degree of the enhancement varied with the species of amino acid and salt. The responses to most amino acids were enhanced by sodium, potassium, and calcium salts, but not enhanced by magnesium salts. The response to methionine, for example, was enhanced by NaCl, but not by Na phosphate of equimolar Na+. 3) The responses to glycine and alanine were suppressed by high concentrations of NaCl. 4) The presence of salts (NaCl and CaCl2) enhanced the responses to amino acids without affecting the thresholds for the amino acids, suggesting that the presence of the salts did not change the affinity of amino acids to the receptor sites. 5) The enhancing effects of salts on the responses to amino acids could not be explained in terms of permeability of cation and anion of salts. It was speculated that the binding of cation and anion of salts on the receptor membranes induces exposure of the receptor sites for amino acids available for binding of amino acids.
Effects of a novel orally effective V1-receptor antagonist, OPC-21268, on AVP-induced sympathoinhibition
Authors: Masaki H. , Imaizumi T. , Harada S. , Momohara M. , Hirooka Y. , Takeshita A.
Published: December 2017

Arginine vasopressin (AVP) plays as important role in control of circulation and may be involved in pathophysiology of cardiovascular diseases. Recently a novel oral V1-receptor antagonist, OPC-21268, has been produced for possible human therapeutic use (Y. Yamamure et al. Science Wash. DC 252: 572-574, 1991). OPC-21268 is a nonpeptide antagonist and inhibits AVP-induced vasoconstriction in rats and humans. In this study, we examined the influence of OPC-21268 on the sympathoinhibitory and bradycardic effects of AVP in conscious rabbits. Before OPC-21268, AVP at doses of 1, 3, 10, and 30 mU/kg decreased renal sympathetic nerve activity (RSNA) by 24 +/- 5, 40 +/- 5, 65 +/- 6 and 86 +/- 5%, respectively, and decreased heart rate from 243 +/- 10 beats/min at control to 232 +/- 11, 221 +/- 10, 197 +/- 9, and 166 +/- 6 beats/min, respectively (n = 12). Oral OPC-21268 at 30 mg/kg (n = 12) and 90 mg/kg (n = 5) did not alter the baseline values of arterial pressure, heart rate, and RSNA. After oral OPC-21268 the decreases in RSNA and heart rate evoked with AVP at graded doses were partially but significantly attenuated (P < 0.01). The attenuation of the AVP-induced decreases in RSNA and heart rate was similar between the two doses of OPC-21268. In another group of rabbits (n = 6), intravenous OPC-21268 at 30 mg/kg almost completely abolished AVP-induced decreases in heart rate and RSNA.(ABSTRACT TRUNCATED AT 250 WORDS)
Vasopressin release within the ventral septal area of the rat brain during drug-induced antipyresis
Authors: Wilkinson M. F. , Kasting N. W.
Published: December 2017

The techniques of push-pull perfusion and radioimmunoassay were used to determine concentrations of arginine vasopressin (AVP) in extracellular fluid derived from the ventral septal area (VSA) of the rat brain following antipyresis elicited by acetaminophen or indomethacin in conscious and unrestrained rats. Reduction of bacterial lipopolysaccharide (LPS)-induced fever by intraperitoneal indomethacin resulted in significant increases in AVP levels in VSA perfusion fluid (P < 0.05). In contrast, antipyresis after acetaminophen treatment was without significant effect on AVP output from VSA nerve terminals. In control animals (non-pyrogen treated), body temperature rose in apparent response to the perfusion procedure. Despite this elevation in core temperature, subsequent treatment with acetaminophen or indomethacin did not result in significant changes in AVP release from VSA perfusates. We conclude that AVP release into VSA extracellular fluids following intraperitoneal indomethacin is dependent upon the neuronal sequelae inherent to pyrogen-evoked fever and not nonspecific rises in body temperature. These results support the hypothesis that endogenous AVP, acting within the VSA, participates in the neuronal mechanisms mediating indomethacin-induced antipyresis.
Caudal hindbrain participation in the suppression of feeding by central and peripheral bombesin
Authors: Ladenheim E. E. , Ritter R. C.
Published: December 2017

We have previously demonstrated that bombesin (BBS) infused into the fourth cerebral ventricle in rats suppressed feeding at doses 10-1,000 times less than those required after lateral ventricular administration. These results suggest that centrally administered BBS acts in the caudal hindbrain to suppress food intake. To investigate the participation of caudal hindbrain structures in fourth-ventricular BBS-induced suppression of feeding, we made lesions that destroyed the following: 1) area postrema (AP) and 2) and medial nucleus of the solitary tract (NST). Additionally, we examined the role of the AP/NST in the suppression of food intake after peripheral BBS administration. Our experiments revealed that lesions of the AP/NST abolished the suppression of food intake by fourth-ventricular BBS and attenuated the suppression of food intake by peripheral BBS. Lesions restricted to the AP attenuated the suppression of food intake by fourth-ventricular BBS but had no effect on suppression of intake by peripherally administered BBS. These results suggest that the NST may be a common neural substrate for the inhibition of food intake by both centrally and peripherally administered BBS.
Norepinephrine does not stimulate protein and UCP synthesis in brown adipocytes of golden Syrian hamsters
Authors: Desautels M. , Dulos R. A.
Published: December 2017

Hamster brown adipocytes were incubated for up to 24 h with or without norepinephrine (NE) in Dulbecco's modified Eagle's medium supplemented with bovine serum albumin, calf serum, and antibiotics. Brown fat cells were viable for 24 h as defined by their ability to respond to NE by a 10-fold increase in oxygen consumption. However, prolonged exposure of the cells to NE led to a decline in NE-stimulated rates of O2 consumption, which was not the result of loss of cell thermogenic capacity. Brown fat cells incubated for 24 h with or without NE showed no significant change in succinate dehydrogenase activity or uncoupling protein (UCP) content. However, cell recovery after 24 h was significantly reduced in the absence of NE. In brown adipocytes isolated from rat, NE increased [35S]methionine incorporation into cell proteins and UCP. In contrast, [35S]methionine incorporation in hamster brown adipocyte proteins and UCP was greater than in rat brown fat cells and was not increased by NE. These results indicate that although NE may be required for cell survival, it does not stimulate protein and UCP synthesis in hamster brown fat cells.
Single-pass gill extraction and tissue distribution of atrial natriuretic peptide in trout
Authors: Olson K. R. , Duff D. W.
Published: December 2017

A new method is described in the present experiments to quantify atrial natriuretic peptide (125I-ANP) extraction from plasma in a single transit through the gill vasculature of an unanesthetized trout. Tissue distribution of 125I relative to an inert extracellular space marker, 58Co-EDTA, was also measured 1 h after injection of 125I-ANP. Single-pass extraction of 125I-ANP by the gill was 60% in control fish; it fell to 18% in fish previously injected with saline and became negative (relative to 58Co-EDTA) after treatment with the clearance (C-type) receptor inhibitor SC 46542. Approximately 90% of a 125I-ANP bolus injected into control trout is removed from the circulation within 2 min. 125I concentration in gill tissue was nearly seven times greater than that predicted by the 58Co-EDTA space. 125I was also concentrated in brain (2X), gallbladder, gastrointestinal tract, and eye (all 1X). SC 46542 decreased gill 125I binding from seven times to one times the 58Co-EDTA space and increased 125I accumulation in heart, kidney, fat, and skeletal muscle. Extraction of 125I-ANP by the isolated perfused gill was 53%, similar to that observed in vivo. These results show that the gill is a major site for ANP removal from trout plasma and that C-type receptors are predominant in this process.
Energy balance and lipid metabolism in transgenic mice bearing an antisense GCR gene construct
Authors: Richard D. , Chapdelaine S. , Deshaies Y. , Pepin M. C. , Barden N.
Published: December 2017

Energy balance and lipid metabolism were investigated in transgenic mice bearing an antisense glucocorticoid receptor (GCR) gene construct that impairs the normal expression of the GCR gene. Food intake was recorded during the 15 days preceding decapitation of adult normal and transgenic mice, and feces were collected to derive the digestible energy intake. Body composition measurements consisted of the determination of energy, protein, and fat content of the carcass. Carcass energy was determined by bomb calorimetry, whereas carcass protein was measured by the Kjeldahl procedure. Energy expenditure was estimated from the continuous oxygen consumption (VO2) monitoring over a 24-h period. Lipoprotein lipase (LPL) activity was quantified in epididymal white adipose tissue (WAT), heart, and vastus lateralis muscle (VLM) by measuring the in vitro hydrolysis of labeled triolein in the presence of tissue homogenates. Norepinephrine (NE) content of both interscapular brown adipose tissue (BAT) and heart were determined by high-performance liquid chromatography (HPLC). Energy intake and expenditure were significantly lower in transgenic mice than in controls. Concurrently, both fat content and total energy of the carcasses were significantly higher in the transgenic animals. In comparison with normal mice, heart and VLM LPL activity was significantly reduced in transgenic mutants. There was no difference between groups in LPL activity in WAT. Finally, heart and BAT NE contents were lower in transgenic animals than in control mice. These results suggest that a defective GCR system may affect energy balance through increasing energetic efficiency, and they emphasize the modulatory effects of hypothalamic-pituitary-adrenal axis changes on muscle LPL activity.
Effects of nifedipine and Bay K 8644 on stimulation-induced adrenal catecholamine secretion in the dog
Authors: Gaspo R. , Yamaguchi N. , De Champlain J.
Published: December 2017

The effects of nifedipine and BAY K 8644 on the adrenal medullary secretion in response to direct splanchnic nerve stimulation were studied in anesthetized dogs. Supramaximal stimulation (12 V) was given on the left splanchnic nerve at a frequency of 2 Hz with three different pulse durations (0.2, 2, and 20 ms) for a total period of 1.5 min. Each stimulation was given for 30 s without interruption between each stimulation. Plasma concentrations of epinephrine and norepinephrine were measured in adrenal venous and aortic blood. In the vehicle control group, epinephrine and norepinephrine concentrations in adrenal venous blood proportionally increased with the lengthening of the pulse duration without significant changes in catecholamine concentrations in aortic blood. In dogs receiving nifedipine (100 micrograms/kg iv), the net increase in adrenal venous epinephrine concentration during stimulation with 20-ms pulse duration was attenuated by approximately 50% (P < 0.05). In dogs treated with BAY K 8644 (30 micrograms.kg-1.min-1 iv), both adrenal venous epinephrine and norepinephrine secretions evoked by stimulation with 20-ms pulse duration were significantly enhanced by approximately 50%. The present results suggest that the secretion of adrenal catecholamines under in vivo conditions is controlled through mechanism(s) involving dihydropyridine sensitive L-type Ca2+ channels presumably localized on the surface of adrenal medullary chromaffin cells.
DPC blockade of transepithelial chloride absorption and single anion channels in teleost urinary bladder
Authors: Chang W. , Loretz C. A.
Published: December 2017

The columnar cell epithelium of the euryhaline goby (Gillichthys mirabilis) urinary bladder actively absorbs NaCl from the lumen, thereby driving water transport and reducing water loss to the hypertonic external environment. Transcellular transport of Cl- involves apical membrane entry via Na(+)-coupled cotransport driven by the Na+ electrochemical gradient and subsequent basolateral membrane exit. An anion channel in the basolateral cell membrane of columnar epithelial cells was identified using patch-clamp technique. This channel may be one avenue for basolateral Cl- exit from the urinary bladder columnar cell. Single-channel conductance (Gc) of channels in excised, inside-out membrane patches was approximately 75 pS in symmetrical solutions containing 140 mM Cl-. The channel was selective to Cl- over other anions [Cl- > 2-(N-morpholino)ethanesulfonic acid (MES) > F- approximately Br- approximately I- > NO3- approximately SO4(2-)). Channel activity, expressed as the open probability (Po), was voltage dependent in the physiological range of membrane potential, with membrane depolarization increasing Po. Decreasing the pH of the solution bathing the cytoplasmic face of the membrane patch over the range 8.4-6.0 reduced Po. There was no effect of pH on either Gc or ionic selectivity. Radiochloride flux technique was also applied to intact columnar epithelial cell sheets to relate anion channel activity to macroscopic transcellular transport. Serosal exposure to the anion channel blocker diphenylamine-2-carboxylic acid (DPC, 30 microM to 3 mM) reduced and abolished transcellular radiochloride fluxes and net Cl- absorption across short-circuited tissues in a dose-dependent fashion. DPC addition (10 microM to 1 mM) to the solution bathing the cytoplasmic face of excised, inside-out membrane patches reduced Po in a dose-dependent manner and had no effect on Gc. These parallel findings of DPC blockade on intact epithelia and on single anion channels support the notion that this anion channel is a basolateral membrane component of the pathway for Cl- movement in transcellular Cl- absorption.
Effect of elevated atrial pressure on plasma renin activity in hypotensive fetal lambs
Authors: Sheikh A. U. , Washburn L. K. , Jaekle R. K. , Rose J. C.
Published: December 2017

In adults, renin secretion is stimulated by reductions in arterial pressure and inhibited by increases in atrial pressure. In the late gestation fetus, a fall in arterial pressure stimulates renin secretion, but it is unknown whether elevation of atrial pressure will alter such an increase. Therefore we studied the effect of elevated atrial pressure on renin secretion in the presence of nitroprusside-induced arterial hypotension. Thirteen fetal lambs at 127.9 +/- 0.9 days of gestation were prepared 5 days before study with inflatable pulmonary artery occluders and right atrial, vascular, and amniotic catheters. Each fetus underwent two protocols (hypotension and hypotension with occlusion) using a randomized block design. Nitroprusside reduced arterial pressure by 34% in both groups. Right atrial pressure during the course of hypotension was significantly higher in the occlusion group (F = 14.2, P = 0.001). Plasma renin activity increased similarly in both groups during hypotension (F = 6.0, P = 0.003). Elevated right atrial pressure did not alter hypotension-induced renin secretion in the fetus.
Effect of outflow pressure on lymphatic pumping in vitro
Authors: Eisenhoffer J. , Elias R. M. , Johnston M. G.
Published: December 2017

The objective of this study was to determine how lymphatic vessels responded to outflow pressure changes in vitro. Bovine mesenteric lymphatics were suspended in an organ bath preparation with both inflow and outflow ends cannulated. Input to the duct was provided from a reservoir filled with Krebs solution. To initiate pumping, a transmural pressure was applied to the ducts by elevating the fluid reservoir and outflow catheters and making their heights equal to one another. The outflow catheter was then elevated above the liquid in the reservoir in 2-cmH2O increments, and pumping activity was monitored for 10 min at each outflow pressure. Outflow pressures were calculated as the product of the flow rate and outflow cannula resistance plus the height of the tip of the outflow catheter above the liquid in the organ bath. At low transmural pressures (2-4 cmH2O), elevations in outflow pressure often had little effect on flow rates until high outflow pressures had been attained. In contrast, elevations in outflow pressures resulted in an increasingly rapid decline in flow rates as transmural pressures were incrementally increased. The mean power (in mu W) required to produce the observed flow rate was estimated at each outflow pressure as the product of the flow rate and the pressure across the lymphatic vessel. The ability of the lymphatics to generate sustained or enhanced power output in response to an increasing outflow pressure challenge was most pronounced at lower transmural pressures. As transmural pressures were increased, the range of outflow pressures that stimulated increased power production was diminished. We conclude that elevations in outflow pressure in an in vitro preparation result in a nonlinear decline in flow rates. This nonlinearity is due to an active lymphatic pump mechanism.
Laryngeal afferent inputs to the nucleus of the solitary tract
Authors: Mifflin S. W.
Published: December 2017

The following study was undertaken to examine the integration of laryngeal afferent inputs within the nucleus of the solitary tract (NTS), the primary site of termination of laryngeal afferent fibers. Intracellular recordings were obtained from 63 cells that responded to electrical stimulation of the superior laryngeal nerve (SLN) with an excitatory postsynaptic potential (EPSP; n = 49), an excitatory-inhibitory postsynaptic potential (EPSP-IPSP) sequence (n = 13), or an IPSP (n = 1). Mechanical stimulation of laryngeal mechanoreceptors revealed a variety of response patterns (e.g., slowly and rapidly adapting depolarizations or hyperpolarizations). Two types of response to increasing SLN stimulus frequency were observed. In 11 cells SLN-evoked EPSP amplitude at 10 Hz was only 47 +/- 4% of the amplitude at 1 Hz, while in 6 cells EPSP amplitude at 10 Hz was virtually identical (93 +/- 3%) to that at 1 Hz. Time-dependent inhibitory interactions occurred between SLN inputs to NTS neurons at intervals between 50 and 400 ms and in the absence of any change in membrane potential. NTS neuronal responses to brief activation of laryngeal mechanoreceptors correspond well to discharge patterns described for individual laryngeal mechanoreceptors. Frequency-dependent filtering and time-dependent inhibitory interactions might modify NTS neuronal responses during more intense stimulation of laryngeal afferents.
Experimental cachexia: effects of MCA sarcoma in the Fischer rat
Authors: Smith B. K. , Conn C. A. , Kluger M. J.
Published: December 2017

Temporal patterns of the cachectic effects of tumor growth and their relation to systemic levels of tumor necrosis factor (TNF) and IL-6 (interleukin-6) were examined in a rat model of experimental cancer cachexia employing the methylcholanthrene (MCA) sarcoma. Fischer 344 rats, implanted with biotelemeters for measuring temperature and activity, were implanted subdermally with tumor tissue fragments. Ad libitum-fed and pair-fed controls were sham incised. Bioassays for TNF and IL-6 were performed on serial plasma samples, obtained via jugular vein at 3- to 6-day intervals throughout the experimental period. Tumor growth induced significant anorexia, weight loss, and a decline in motor activity corresponding to an increase in mean plasma IL-6 levels, independent of reduced food intake or weight loss alone as shown in pair-fed controls. A significant lowering of body temperature then developed, followed by a two- to threefold increase in water consumption. The patterns of weight loss and temperature reduction differed in rate and degree from those seen with pair feeding.
Central interferon-alpha inhibits natural killer cytotoxicity through sympathetic innervation
Authors: Take S. , Mori T. , Katafuchi T. , Hori T.
Published: December 2017

The brain has been known to produce high levels of interferon-alpha (IFN-alpha) during viral infections. We investigated the central and peripheral mechanisms of the brain IFN-alpha-induced suppression of natural killer (NK) cytotoxicity in the rat. The activity of NK cells in the spleen and the peripheral blood decreased 30-120 min after intracerebroventricular (icv) injection of recombinant human IFN-alpha of > 1,000 U but not after its intraperitoneal injection. This effect was antagonized by pretreatment with icv naltrexone (NLTX). Splenic denervation was observed to completely abolish the IFN-alpha-induced suppression of NK activity, whereas bilateral adrenalectomy did not. Furthermore, this immunosuppression was blocked by an icv injection of an antagonist of corticotropin-releasing factor (CRF), alpha-helical CRF-(9-41). The icv injection of CRF resulted in reduced NK activity, which was not affected by NLTX. The results suggest that brain IFN-alpha activates the CRF system through central opioid receptors and thereby suppresses the NK cytotoxicity predominantly through splenic sympathetic innervation.
Vasopressin-independent alterations in renal water excretion in quadriplegia
Authors: Wall B. M. , Williams H. H. , Presley D. N. , Crofton J. T. , Share L. , Cooke C. R.
Published: December 2017

Postural effects on water excretion are known to be increased in patients with cervical spinal cord injury and may result in marked impairment of the ability to excrete a water load, especially in erect posture. Both vasopressin-dependent and vasopressin-independent mechanisms have been implicated. To assess the roles of these mechanisms and further identify the factors involved in the renal response to erect posture, sustained water loading studies were performed on 11 quadriplegic subjects and 9 healthy control subjects, supine and erect (sitting). Renal blood flow was assessed by p-aminohippurate clearance (CPAH) measurements in 7 quadriplegic and 5 control subjects. During maximal water diuresis, plasma vasopressin concentrations were reduced to unquantifiable levels in all subjects. Osmolar clearance, free water clearance (CH2O), and distal delivery of filtrate (DDF) were all lower in quadriplegic than in control subjects, supine and erect. The relationship between CH2O and DDF was the same in quadriplegic as in control subjects and was not altered by change in posture in either group. Creatinine clearance and CPAH were lower in erect than in supine posture in quadriplegic subjects but not in control subjects. We conclude that impairment of water excretion in stable normonatremic quadriplegic subjects can be attributed primarily to vasopressin-independent mechanisms involving reduced filtrate delivery to diluting segments of the renal tubules rather than to resistance to normal suppression of vasopressin release.
High-density localization of ANP receptors in chondrocytes of eel gill cartilage
Authors: Sakaguchi H. , Katafuchi T. , Hagiwara H. , Takei Y. , Hirose S.
Published: December 2017

Very high levels of atrial natriuretic peptide (ANP) receptor were found in a quite unexpected tissue, i.e., the chondrocyte of the eel gill cartilage. 125I-labeled ANP binding assay indicated that the eel gill contains large amounts of the receptor and its levels are much higher (> 10-fold) than those in the other tissues examined, including the brain, kidney, head kidney including steroidogenic interrenal cells, and gut. Autoradiography using gill sections revealed dense localization of ANP receptors in the chondrocytes of the cartilage as well as moderate localization in the parenchymal cells. Biochemical and pharmacological characterization of the chondrocyte receptor indicated that most of the receptors are type C receptors, having a broad ligand specificity and a reduced M(r) of 68,000. Because a hyaline cartilage in which chondrocytes reside lacks blood vessels, and all metabolic exchange is by slow diffusion through the matrix, the chondrocyte localization of the type C ANP receptor is notable and may stimulate not only chondrocyte research but also studies on physiological significance of the type C receptor, which is generally considered to be merely a clearance (silent) receptor.
Skeletal muscle Ca2+ flux and catabolic response during sepsis
Authors: Bhattacharyya J. , Thompson K. D. , Sayeed M. M.
Published: December 2017

Membrane Ca2+ flux and net protein catabolism were studied in the skeletal muscle during experimental sepsis. Sterilized rat fecal pellets with (septic) or without (sterile) gram-negative bacteria, Escherichia coli [10(2) colony-forming units (cfu)] and Bacteroides fragilis (2 x 10(3) cfu), were implanted into the abdomens of male Sprague-Dawley rats (110-120 g). Septic and sterile rats were febrile and hyperlactacidemic on day 1 postimplantation. These responses subsided by day 2 in sterile but not septic rats. Initial Ca2+ flux, estimated from measurements of 45Ca uptake by soleus muscles in vitro, was elevated on day 1 in both sterile and septic rats and on day 2 and 3 in septic rats only. The septic rat soleus muscle showed a significantly increased net protein catabolic response (measured as tyrosine release by soleus muscle, in vitro) over that found in muscles of sterile rats on day 1-3 postimplantation. The increase in Ca2+ flux in septic (day 1-3 postimplantation) and sterile (day 1 only) rats was abolished when the rats were treated with the calcium channel blocker diltiazem. In unoperated control rat soleus muscles the Ca2+ ionophore, ionomycin, concomitantly caused an increase in Ca2+ flux and net protein catabolism. Overall, the present study suggested that altered cellular Ca2+ regulation plays a role in the net protein catabolic response in the skeletal muscle during sepsis.
Differential timing of amino acid and 5-HIAA rhythms in suprachiasmatic hypothalamus
Authors: Glass J. D. , Hauser U. E. , Blank J. L. , Selim M. , Rea M. A.
Published: December 2017

In vivo brain microdialysis was used to characterize the daily pattern of extracellular excitatory amino acids (EAA; glutamate and aspartate), glutamine and 5-hydroxyindoleacetic acid (5-HIAA) in the region of the suprachiasmatic nucleus (SCN) in freely behaving male Syrian hamsters. Diurnal variations in the extracellular concentrations of EAA and 5-HIAA observed under 14:10-h light-dark (LD) photoperiod were confirmed by cosinor analysis. Peak levels occurred during the night, with the mean acrophasis for 5-HI-AA concentration preceding that for EAA by 1-2 h. Release of EAA was stimulated by K(+)-induced depolarization in a Ca(2+)-dependent manner and was not affected by tetrodotoxin. In the absence of light cues (constant dim red light, < 0.4 lx; DD) there was a significant time-of-day effect in peak glutamate concentration (occurring during subjective night), but the rhythmic pattern of 5-HIAA output was lost. There was no apparent direct temporal linkage between peaks in extracellular EAA or 5-HIAA and bouts of wheel-running activity under LD or DD. These results are evidence that 1) the daily release pattern of extracellular glutamate, but not 5-HIAA, is circadian in nature; and 2) this rhythm in glutamate is not based on Na+ channel-dependent action potentials. Finally, a discordance in the timing of peak nocturnal extracellular 5-HIAA and EAA concentrations was evident, possibly reflecting an interplay between serotonergic and EAA pathways in the SCN.
Role of hypothalamic interleukin-6 and tumor necrosis factor-alpha in LPS fever in rat
Authors: Klir J. J. , Roth J. , Szelenyi Z. , McClellan J. L. , Kluger M. J.
Published: December 2017

The purpose of this study was to determine, using push-pull perfusion, the levels of interleukin (IL)-1-like, IL-6-like, and tumor necrosis factor-alpha (TNF)-like activity in the anterior hypothalamus during lipopolysaccharide (LPS)-induced fever in rats. Additionally, slow anterior hypothalamic infusions of human recombinant IL-6 (hrIL-6) or TNF (hrTNF) for several hours were performed to determine possible febrile effects of these two cytokines. Artificial cerebrospinal fluid (aCSF) was infused as a control. Samples of cerebrospinal fluid were collected 60 min before and 60, 180, 300, and 420 min after the intraperitoneal injection of LPS. A control group was injected intraperitoneally with saline. The core temperature (measured by biotelemetry) of LPS-injected rats was significantly higher (P < 0.05) than the temperature of the rats injected with saline at 180, 300, and 420 min after the injection. The average postinjection IL-6 levels were significantly higher (P < 0.05) in the LPS-injected group. TNF was significantly higher (P < 0.05) than the baseline only at 180 min. There were no changes in levels of IL-1-like activity. Infusion of hrIL-6 at a level similar to the peak IL-6 level measured during LPS-induced fever resulted in a slowly developing and long-lasting increase in core temperature. Infusion of hrTNF at a level corresponding to the peak TNF level measured during LPS-induced fever did not induce a significant increase in core temperature. These results support the hypothesis that elevated hypothalamic concentrations of IL-6 are involved in the induction of fever elicited by peripheral (intraperitoneal) injection of LPS.
Nitric oxide and superoxide in cultured cells: limited production and influence on DNA synthesis
Authors: Firnhaber C. , Murphy M. E.
Published: December 2017

The ability of superoxide (O2-) and NO to influence DNA synthesis, as estimated by [3H]thymidine incorporation in logarithmically growing cells, was evaluated in three human cell types: embryonic lung fibroblasts, skin fibroblasts, and mesangial cells. Nontoxic rates of O2- generation (0.2 nM/min-0.2 microM/min) only slightly stimulated DNA synthesis in mesangial cells (10-13% increase) and did not affect fibroblast DNA synthesis. Nontoxic rates of NO generation (0.1-10 microM/min) also had only limited effects, inhibiting DNA synthesis in lung and skin fibroblasts slightly (10-20% decrease) but not affecting mesangial cells. In all three cell types, neither O2- nor NO was generated at high enough endogenous rates to support an autocrinic regulation of DNA synthesis. This and other lines of evidence indicate that a reaction between O2- and NO is unlikely to account for their opposing effects on DNA synthesis.
Anti-TNF-alpha antibodies normalized body temperature and enhanced food intake in tumor-bearing rats
Authors: Smith B. K. , Kluger M. J.
Published: December 2017

Anorectic weight-losing Fischer 344 rats bearing a methylcholanthrene-induced sarcoma received intraperitoneal injections of rabbit antiserum raised against murine tumor necrosis factor (TNF)-alpha at 14, 18, and 24 days after tumor induction. Treatment of tumor-bearing rats with TNF antiserum partially reversed the tumor-induced reduction in food intake compared with tumor-bearing rats that received control serum. In the same tumor-bearing animals, treatment with TNF antiserum delayed the onset and significantly reduced the decline in mean 12-h daytime and nighttime intra-abdominal temperatures on days 18-25. However, anti-TNF antibody treatment did not alter the declines in carcass weight or motor activity measured from day of tumor induction until death or reduce the tumor burden at death. We conclude that an endogenous TNF response may be one of the factors involved in the development of cancer anorexia and that this cytokine has temperature-lowering properties.
Kinetics of systemic and intrahypothalamic IL-6 and tumor necrosis factor during endotoxin fever in guinea pigs
Authors: Roth J. , Conn C. A. , Kluger M. J. , Zeisberger E.
Published: December 2017

The time course of activity of interleukin-6 (IL-6) and tumor necrosis factor (TNF) was measured in blood plasma and hypothalamic push-pull perfusates during the febrile response to intramuscular injection of bacterial endotoxin (Escherichia coli, 20 micrograms/kg) in 24 guinea pigs. Injection of endotoxin caused a dramatic increase of IL-6 activity in plasma. The logarithmic values of plasma IL-6 activities showed a linear correlation to the febrile change in body temperature (r = 0.898) during the whole time course of fever. IL-6 activity in hypothalamic perfusates increased 12-fold in the first hour after pyrogen application and declined slowly despite the further increase in body temperature. Hypothalamic IL-6 activity did not correlate with the febrile increase in body temperature (r = -0.048). TNF activity in plasma, not detectable before pyrogen application, had its peak in the first hour after endotoxin injection and rapidly declined to 15-20% of the peak activity within the next 2 h and to an undetectable value 5 h after injection. In the hypothalamus TNF was not detectable before endotoxin injection, but it could be monitored in most animals after pyrogen application without a clear correlation to the fever response. These results taken together indicate that endotoxin fever represents a physiological situation in which production and release of cytokines in the peripheral immune system and in the hypothalamus are regulated and stimulated in independent patterns.