bluuebunny/crossref_metadata_2025 huggingface dataset:
A method for calculating the distribution of pH in tissues and a new source of pH error from the 31P-NMR spectrum
Authors: Graham R. A. , Taylor A. H. , Brown T. R.
Published: December 2017

The true distribution of the pH in tissues can be determined from the in vivo 31P-nuclear magnetic resonance (NMR) spectrum by converting the parts per million (PPM) axis of the pH responsive resonance to pH using the Henderson-Hasselbalch equation. In addition, the intensity axis of the resonance must be divided by the derivative of the Henderson-Hasselbalch equation to correct for the nonlinear relationship between pH and PPM. This nonlinear relationship causes the apparent center of the resonance in PPM to be dependent not only on the center of the pH distribution but also on its width and distance from the pKa, where Ka is the association constant. Therefore, the pH determined from uncorrected spectra may be in significant error, particularly if the pH distribution is distant from the pKa and is broad. The method was applied to the isolated perfused Morris hepatoma 5123C to determine the distribution of intracellular pH (pHi) using resonances from two intracellular compounds. The two resonances did not report the same pHi unless the spectral data were properly corrected. The method should be of interest to anyone interested in pHi.
Cerebral metabolism and electrocortical activity in the chronically hyperglycemic lamb
Authors: Rosenkrantz T. S. , Knox I. , Zalneraitis E. L. , Raye J. R. , Porte P. J. , Cramer R. , Smoloski R. , Philipps A. F.
Published: December 2017

Pages R1262–R1269: T. S. Rosenkrantz, I. Knox, E. L. Zalneraitis, J. R. Raye, P. J. Porte, R. Cramer, R. Smoloski, and A. F. Philipps. “Cerebral metabolism and electrocortical activity in the chronically hyperglycemic lamb.” Page R1265: Several errors relating to significance of changes in arterial oxygen concentration, cerebral oxygen delivery, and cerebral oxygen uptake were introduced into Table 1 by the publisher. All of the information in the abstract and text are correct as originally printed. A corrected version of the table is reprinted below. Note that there were no changes in the fetal arterial oxygen content during any of the study periods, cerebral oxygen delivery rose during G2 ( P = 0.06), and there was a significant increase in cerebral oxygen uptake during G2 ( P < 0.05). It is hoped that this clarifies conflicting information in the text and the table concerning the metabolic changes that occur in the brain of the chronically hyperglycemic fetal lamb.

(See PDF)
Membrane fluidity and hemilayer temperature sensitivity in trout hepatocytes during brief in vitro cold exposure
Authors: Williams E. E. , Hazel J. R.
Published: December 2017

Fluorescent membrane probes were used to assess the fluidity of hepatocyte plasma membranes (PM) from 20 degrees C-acclimated trout after exposure to 20 and 5 degrees C. PM isolated from cells after 6 h at 5 degrees C were significantly more fluid [fluorescence depolarization of 1,6-diphenyl-1,3,5-hexatriene (DPH)] than control membranes at both temperatures. The increased fluidity was sufficient to offset 45-50% of the cold-induced membrane ordering. In contrast, the fluidity of PM in intact cells from 20 degrees C-acclimated fish remained constant when exposed to 5 degrees C for a similar period. In addition, the fluidity of the inner hemilayer [1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene, p-toluenesulfonate (TMA-DPH)] was significantly less sensitive to temperature change than was the fluidity of the outer hemilayer [3-(p-(6-phenyl)-1,3,5-hexatrienyl)phenylpropionic acid (PA-DPH)]. Because the isolated membrane preparation was most likely enriched with canalicular membranes (based on 5'-nucleotidase recovery), these results suggest that the canalicular domain of the plasma membrane is preferentially modified during short-term cold exposure and that the fluidity of the inner hemilayer of the plasma membrane of intact cells is relatively temperature insensitive, thus requiring fewer modifications than the outer hemilayer during temperature acclimation.
Cardiac performance in semi-isolated heart of the crab Carcinus maenas
Authors: Wilkens J. L. , McMahon B. R.
Published: December 2017

A semi-isolated, in situ heart preparation of the shore crab, Carcinus maenas, supported by its alary ligaments, pumps vigorously for hours at a mean heart rate of 49.7 beats/min and cardiac output of 30 ml.kg-1.min-1. These hearts show no adaptive responses to changes in pericardial sinus pressure, outflow resistance, or afterload. Direct perfusion-induced stretch of the heart wall causes increases in contractile force but minimal changes in heart rate. Stroke work and power are lower than comparable values for animals with myogenic hearts and closed circulatory systems. The values for heart rate and cardiac output are lower than in vivo values and may in part reflect the technique used as well as intrinsic performance of the heart without neural and neurohormonal inputs. Morphometrically the heart represents 0.2% of whole body weight, and the mean stroke volume of 0.35-0.45 ml/kg represents an ejection fraction of 27-34% of ventricular volume (1.4 ml/kg).
Regulation of plasma vasopressin by plasma osmolality and carotid sinus pressure in anesthetized rabbits
Authors: Scott C. S. , Sharp-Kehl J. , Redekopp C. A. , Ledsome J. R.
Published: December 2017

The purpose of the experiments was to investigate the effect of changes in carotid sinus baroreceptor stimulation on plasma vasopressin (AVP) at different plasma osmolalities in the anesthetized artificially ventilated rabbit. Both carotid sinuses were isolated and perfused with blood at servo-controlled pressures. The vagus and aortic depressor nerves were sectioned bilaterally to eliminate input from atrial and aortic arch baroreceptors. Saline (0.3%, wt/vol) was infused to lower plasma osmolality, and 5% saline was infused to raise plasma osmolality. At three plasma osmolalities, the carotid sinus pressure (CSP) was changed from 100 mmHg to 40 and 140 mmHg and returned to 100 mmHg. There were no changes in plasma AVP in response to changes in CSP at low plasma osmolality (289 mosmol/kgH2O), but at medium (309 mosmol/kgH2O) and high (323 mosmol/kgH2O) osmolality, plasma AVP was higher at 40 than at 140 mmHg CSP. The relationship between plasma AVP and plasma osmolality was expressed as a linear regression at each CSP. Changes in CSP changed the sensitivity but not the threshold of the osmotic control of AVP release.
Role of mast cell histamine in brown adipose tissue thermogenic response to VMH stimulation
Authors: Desautels M. , Wollin A. , Halvorson I. , Muralidhara D. V. , Thornhill J.
Published: December 2017

Electrical stimulation of the ventromedial hypothalamic area in rats caused a significant but transient increase in interscapular brown adipose tissue temperature. This response was markedly reduced by cimetidine, a histamine H2-receptor antagonist, but not by pyrilamine, an H1-receptor antagonist. Histamine is present in substantial amounts within mast cells in brown adipose tissue as injections of compound 48/80, which cause degranulation of connective tissue mast cells, reduced the tissue histamine content by > 85%. In contrast, histamine content in brown adipose tissue was not affected by loss of sympathetic neural input (with 6-hydroxydopamine) or sensory neural input (with capsaicin). Neither cimetidine nor histamine had any effect on basal and norepinephrine-stimulated rates of O2 consumption by isolated brown adipocytes. These results indicate that histamine released from mast cells acting on H2-receptors may play an important but indirect role in the thermogenic response of brown adipose tissue to stimulation of the ventromedial hypothalamic area.
Chloride-dependent intestinal transport of imino and beta-amino acids in the guinea pig and rat
Authors: Munck L. K. , Munck B. G.
Published: December 2017

The present study extends the observations of chloride-dependent intestinal amino acid carriers to the guinea pig and the rat using the technique of in vitro influx across the brush-border membrane of intact epithelium. Transport rates of D-glucose and L-amino acids are lowest in guinea pig proximal small intestine and are constant from midjejunum through distal ileum, except for leucine. The guinea pig possesses a sodium- and chloride-dependent, high-affinity, very low-capacity carrier of beta-amino acids for which taurine and beta-alanine compete and for which the Na(+)-taurine activation stoichiometry is 2.1 +/- 0.3:1. The imino acid carrier of the guinea pig is also chloride dependent with a Na(+)-Cl(-)-2-methylamino-isobutyric acid activation stoichiometry of 1.8 +/- 0.1:0.7 +/- 0.3:1. In contrast, the rat imino acid carrier is chloride independent and transport rates vary insignificantly along the small intestine. The rat taurine carrier has its maximal transport rate in midjejunum. It is chloride dependent but does not transport beta-alanine.
Response of the magnocellular system in rats to hypovolemia and cholecystokinin during pregnancy and lactation
Authors: Koehler E. M. , McLemore G. L. , Martel J. K. , Summy-Long J. Y.
Published: December 2017

To define changes in the magnocellular neuroendocrine system during lactation and pregnancy, we compared plasma levels of oxytocin (OT) and vasopressin (VP) after polyethylene glycol (PEG)-induced hypovolemia and cholecystokinin (CCK) stimulation. Conscious virgin, pregnant (day 20), and lactating (day 6) Sprague-Dawley rats were injected with either PEG (70-600 mg/ml; 35 or 70 ml/kg sc), CCK (100 micrograms/ml; 4 ml/kg ip), or vehicle and decapitated 4 h (PEG) or 5 min (CCK) later. Changes in thresholds for release of hormone and the responsiveness (slopes relating [hormone] to blood volume depletion or to plasma osmolality) of the OT and VP systems were determined using an iterative nonlinear threshold regression model. After PEG, plasma osmolality increased coincident with a decrease in blood volume, with both stimuli contributing to the rise in plasma VP and OT. Compared with virgin rats, neither the threshold nor the responsiveness of the VP system was altered by the combined stimulus, whereas the oxytocinergic system of pregnant rats was more responsive to osmotic component. Lactating rats, however, had a higher threshold for VP release and an apparent elevation of the OT threshold beyond 25% volume depletion. Regardless of the reproductive state, the threshold for VP release was always lower than that for OT. Intraperitoneal CCK elevated plasma [OT] in each reproductive state, although the response in lactating animals was attenuated. In virgin and lactating rats, plasma levels of VP also increased slightly but significantly in response to CCK. During gestation when cardiovascular volume is expanded, both the VP and OT neuroendocrine systems were reset, enabling secretion of both hormones in response to hypovolemia with hypertonicity. During lactation, both neuroendocrine systems are reset such that greater changes in fluid balance are needed to stimulate hormone release. Regardless of the reproductive state, the threshold for VP release was always lower than that for OT, indicative of preferential release of VP with less than a 5% (virgin, pregnant) or a 20% (lactating) loss in blood volume.
Swallowing and urine flow responses of ovine fetuses to 24 h of hypoxia
Authors: Brace R. A. , Wlodek M. E. , McCrabb G. J. , Harding R.
Published: December 2017

The aim of this study was to determine the effects of 24 h of hypoxia on fetal swallowing and urine flow rates. The study design included successive 24-h control, hypoxia, and recovery periods. To induce hypoxia, we infused nitrogen into the trachea of late-gestation pregnant sheep. During hypoxia, there were decreases in fetal arterial oxygen saturation [from 62.7 +/- 2.2 to 30.9 +/- 2.9% (SE)] and PO2 (from 21.1 +/- 1.0 to 14.1 +/- 0.9 mmHg) (n = 7). Fetal arterial pH decreased maximally by 0.04 +/- 0.01 at 6 h and returned to control thereafter. Fetal swallowing decreased from 13.4 +/- 2.1 to 0.6 +/- 0.2 ml.h-1.kg fetal wt-1 with the onset of hypoxia and gradually increased, but only to one-half control levels, during the last 12 h of hypoxia. Fetal urine flow gradually decreased from 9.6 +/- 1.6 ml.h-1.kg-1 to a minimum of 5.3 +/- 0.5 ml.h-1.kg-1 at 2-3 h of hypoxia and returned to control thereafter. During the first hour of the recovery period, fetal swallowing transiently increased to twice control levels before returning to control. This was followed by a delayed, transient increase in urine flow to 63% above control levels at 2-6 h after hypoxia. We conclude that the fetal swallowing and urine flow responses to prolonged hypoxia in the absence of acidemia are distinctly different from the initial inhibitory responses, in that swallowing partially and urine flow fully recovers from the initial suppression.
Evolution of marsupial and other vertebrate thyroxine-binding plasma proteins
Authors: Richardson S. J. , Bradley A. J. , Duan W. , Wettenhall R. E. , Harms P. J. , Babon J. J. , Southwell B. R. , Nicol S. , Donnellan S. C. , Schreiber G.
Published: December 2017

Binding of radioactive thyroxine to proteins in the plasma of vertebrates was studied by electrophoresis followed by autoradiography. Albumin was found to be a thyroxine carrier in the blood of all studied fish, amphibians, reptiles, monotremes, marsupials, eutherians (placental mammals), and birds. Thyroxine binding to transthyretin was detected in the blood of eutherians, diprotodont marsupials, and birds, but not in blood from fish, toads, reptiles, monotremes, and Australian polyprotodont marsupials. Globulins binding thyroxine were only observed in the plasma of some mammals. Apparently, albumin is the phylogenetically oldest thyroxine carrier in vertebrate blood. Transthyretin gene expression in the liver developed in parallel, and independently, in the evolutionary lineages leading to eutherians, to diprotodont marsupials, and to birds. In contrast, high transthyretin mRNA levels, strong synthesis, and secretion of transthyretin in choroid plexus from reptiles and birds indicate that transthyretin gene expression in the choroid plexus evolved much earlier than in the liver, probably at the stage of the stem reptiles. NH2-terminal sequence analysis suggests a change of transthyretin pre-mRNA splicing during evolution.
Receptor proteins and biological effects of C-type natriuretic peptides in the renal glomerulus of the rat
Authors: Brown J. , Zuo Z.
Published: December 2017

Binding studies on rat glomeruli using 125I-labeled Tyr0-C-type natriuretic peptide-(1-22) [125I-Tyr0-CNP-(1-22)] and 125I-labeled alpha-atrial natriuretic peptide (alpha-125I-ANP), and the unlabeled ligands CNP-(1-22), alpha-ANP, and des-Gln18,Ser19,Gly20,Leu21,Gly22-ANP-(4-2 3)-NH2 (C-ANP) suggest that receptor-like sites that bind both alpha-ANP and C-ANP fall into two categories, one with high [dissociation constant (Kd) approximately 10(-9)M] and one with low (Kd approximately 10(-5)M) affinity for CNP-(1-22). Covalent attachment of 125I-Tyr0-CNP-(1-22) and alpha-125I-ANP to these sites identifies two membrane proteins with corresponding properties. The first, which can be labeled by both radioligands, is a disulfide-bridged approximately 140-kDa protein that is reduced by dithiothreitol to approximately 67 kDa. This protein binds C-ANP and has Kd approximately 10(-10) M for CNP-(1-22). The second protein, which is labeled only by alpha-125I-ANP, also binds C-ANP, but has Kd approximately 10(-5)M for CNP-(1-22). This approximately 77-kDa protein may also have a disulfide-bridged, high-molecular-mass form of approximately 140 kDa in the absence of dithiothreitol. Studies of glomerular function show that alpha-125I-ANP is internalized whereas 125I-Tyr0-CNP-(1-22) is not. C-ANP abolishes the specific internalization of alpha-125I-ANP. CNP-(1-22) inhibits internalization of 400 pM alpha-125I-ANP weakly, only approximately 60% being inhibited by 10 microM CNP-(1-22). This implies that the approximately 77-kDa protein, with its low affinity for CNP-(1-22), mediates internalization. Furthermore, CNP-(1-22), as well as alpha-ANP and C-ANP, inhibits glomerular levels of adenosine 3',5'-cyclic monophosphate (cAMP), and CNP-(1-22) does so with a high affinity, which corresponds to its affinity for the approximately 67-kDa protein. The results suggest that the approximately 67-kDa receptor is distinct from the natriuretic peptide clearance receptor and may control cAMP levels.
Extravascular protein in the renal medulla: analysis by two methods
Authors: Pallone T. L.
Published: December 2017

Two methods have been used to test for the presence of extravascular protein in the interstitium of the renal inner medulla. First, ascending vasa recta (AVR) segments were perfused with buffer containing 5 g/dl of albumin. The hydraulic pressure in the perfused vessel was varied to control transmembrane volume flux (Jv) to the interstitium. Interpolation to the point of zero Jv was employed to estimate interstitial Starling forces in the hydropenic rat papilla. Analysis of those experiments predicts that interstitial protein concentration (Ci) is high. When AVR segments are filled with oil, the oil column spontaneously breaks up as fluid is secreted into the lumen from the papillary interstitium. To obtain a lower limit on Ci, isolated AVR segments (IAS) filled with oil were sampled to measure protein concentration in the secreted fluid. In hydropenic rats, protein concentration was 3.4 +/- 0.5 and 5.2 +/- 0.2 g/dl in IAS and adjacent free-flowing AVR, respectively (P < 0.01). In rats subjected to furosemide and saline diuresis, the values were nearly identical, 4.7 +/- 0.2 and 5.2 +/- 0.2 g/dl, respectively. These separate experimental approaches corroborate a high concentration of protein in the renal inner medullary interstitium.
Effects of ANG-converting enzyme and alpha 1-adrenoceptor inhibition on intrarenal hemodynamics in SHR
Authors: Numabe A. , Komatsu K. , Frohlich E. D.
Published: December 2017

To investigate the prolonged effects of angiotensin-converting enzyme (ACE) inhibition and alpha 1-adrenoceptor blockade on intrarenal hemodynamics, whole kidney and renal micropuncture studies were performed in male 21-wk-old Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats treated for 3 wk with quinapril (3 mg/kg), terazosin (1 mg/kg), or in their combination (quinapril 1.5 mg/kg and terazosin 0.5 mg/kg). In WKY, only quinapril significantly reduced mean arterial pressure (MAP) associated with reduced afferent arteriolar resistance; there were no other significant changes. In contrast, all treatments similarly decreased MAP in SHR. Quinapril increased renal plasma flow and decreased filtration fraction. With respect to intrarenal hemodynamics, quinapril increased single-nephron plasma flow and reduced glomerular capillary pressure (from 53.1 to 47.8 mmHg; P < 0.01), associated with reduced afferent (from 4.80 to 3.17 10(10)dyn.s.cm-5; P < 0.01) and efferent (from 1.70 to 1.17 10(10)dyn.s.cm-5; P < 0.01) arteriolar resistances, and increased ultrafiltration coefficient (from 0.037 to 0.052 nl.s-1.mmHg-1; P < 0.05). Terazosin only reduced arteriolar resistance. The combined treatment attenuated either agent's independent effects on glomerular hemodynamics. These data demonstrate that prolonged ACE and adrenergic inhibition therapy alone or in combination produce different effects than when given by vein, suggesting that prolonged renopressor system inhibition may be more effective than adrenergic in SHR.
Role of endogenous vasopressin in development of gastric ulcer induced by restraint and water immersion
Authors: Honda K. , Fukuda S. , Ishikawa S. E. , Kuzuya T. , Saito T.
Published: December 2017

To elucidate the role of arginine vasopressin (AVP) in the development of stress-induced gastric ulcer, the mucosal lesions after restraint and water immersion were examined in Brattleboro strain rats with hereditary hypothalamic diabetes insipidus (DI) and in Long-Evans rats (LE) used as controls. Restrained animals were immersed in water for 2 h, and the size of lesion was expressed as percentage of the lesion area to the total glandular mucosal area, which were defined as ulcer index (UI). In DI rats, UI was significantly higher than in control LE rats, despite the attenuated responses of plasma adrenocorticotropic hormone (ACTH) to stress. Although subcutaneous injection of selective antidiuretic analogue 1-desamino-8-D-AVP did not affect UI, intracerebroventricular (icv) administration of AVP reduced UI in DI rats, and icv administration of V1 antagonist [d(CH2)5Tyr(Me)]AVP elevated UI in LE rats. These results indicate that endogenous AVP plays a role in preventing the formation of gastric ulcers induced by stress via a central V1 receptor. Furthermore, we suggest that elevation of ACTH in plasma is not essential in the development of stress-induced gastric ulcer in rats.
Circadian rhythms and time course of adaptive sodium and potassium excretion in rats after uninephrectomy
Authors: Aizman R. I. , Rabinowitz L. , Mayer-Harnisch C.
Published: December 2017

The relationship between renal circadian cyclic excretion and renal compensatory adaptation after uninephrectomy for K, Na, and water was studied. Rats in a 12:12-h light-dark environment were given a liquid diet, and urine was collected for 16 days with consecutive 90-min periods. Days 1-4 were control, 5-10 followed a sham operation, and 11-16 followed uninephrectomy. The major findings were 1) the circadian cycles in excretion were virtually unchanged after sham and uninephrectomy; 2) an adaptive increase in excretion of Na, K, and water by the remaining kidney occurred within 90 min after uninephrectomy; 3) after uninephrectomy the distribution of the 24-h increment in excretion for the remaining kidney closely followed the preexisting pattern of circadian excretion for Na but was evenly distributed between light and dark phases for K; and 4) after uninephrectomy the ratio of excretion to intake was unchanged. This is the first study to document the time course of adaptation to uninephrectomy using consecutive brief collections over several days in unanesthetized and undisturbed rats. Adaptation after uninephrectomy occurred essentially immediately; was maintained unchanged; and preserved Na, K, and water homeostasis. Uninephrectomy did not alter the circadian control of excretion.
Effects of acute salt loading on vasopressin mRNA level in the rat brain
Authors: Shoji M. , Kimura T. , Kawarabayasi Y. , Ota K. , Inoue M. , Yamamoto T. , Sato K. , Ohta M. , Funyu T. , Sonoyama T. , et al.
Published: December 2017

To assess the mutual relationship between acute osmotic stimulation and arginine vasopressin (AVP) gene expression, 2 ml/100 g body weight of 0.9 M NaCl was intraperitoneally administered into conscious rats. They were decapitated to collect blood and brain samples before and 15 min and 1, 3, 6, and 9 h after the injection. The total RNA from the hypothalamus or whole brain tissue was used to determine AVP mRNA by Northern blot analyses with a complementary DNA probe. Plasma AVP and osmolality increased rapidly and transiently 15 min and 1 and 3 h after the injection. AVP mRNA was detected in the hypothalamus but not in the brain tissue without hypothalamus under basal and stimulated conditions. Brain AVP mRNA increased 2.2-fold at 3 h and 1.7-fold at 6 h (P < 0.05-0.01). These increases appeared to be due to the appearance of AVP mRNA with the shorter migration in the gel. These results suggest that an acute osmotic challenge increases AVP mRNA with size heterogeneity within the hypothalamo-hypophyseal tract.
Influence of pregnancy on ANF release from isolated atria
Authors: Kaufman S. , Deng Y. , Thai W.
Published: December 2017

Isolated perfused right atria were prepared from virgin cycling rats and from rats at 7, 14, and 21 days of pregnancy. Intraluminal pressure was raised from the basal control level (atmospheric) to 4, 6, and 10 cmH2O. Basal and stretch-induced release of atrial natriuretic factor (ANF) into the perfusate was measured by radioimmunoassay. When atria derived from unmated and from 7-day pregnant rats were distended, mean ANF secretion into the perfusion medium increased by 64 +/- 16 and 89 +/- 31 pg/ml at the highest distending pressure of 10 mmHg. (Mean basal secretion for the two groups was 196 +/- 104 and 181 +/- 63 pg/ml, respectively.) However, at 14 and 21 days, distension failed to elicit any significant increase in ANF release. There was no significant difference between the groups with respect to basal secretion of ANF, nor did pregnancy influence atrial compliance or volume. The slopes of the pressure-volume curves for atria derived from virgin, 7-, 14-, and 21-day-pregnant rats were 0.76 +/- 0.17, 0.623 +/- 0.178, 0.811 +/- 0.177, and 1.050 +/- 0.173, respectively. These results are in agreement with our findings in vivo that plasma ANF levels are elevated at 7 days of pregnancy but that, despite the progressive increase in intravascular volume, they decrease to control levels at 14 and 21 days. Our latest data suggest that this fall is a result of changes in the secretory characteristics of the atrial tissue itself and not necessarily of pregnancy-induced changes in atrial filling.
Role of peripheral capsaicin-sensitive neurons and CGRP in central vagally mediated gastroprotective effect of TRH
Authors: Kato K. , Yang H. , Tache Y.
Published: December 2017

We investigated in conscious rats the role of capsaicin-sensitive neurons and alpha-calcitonin gene-related peptide (CGRP), the form preferentially expressed in capsaicin sensory neurons, in mediating intracisternal thyrotropin-releasing hormone (TRH) analogue-induced vagal muscarinic gastroprotection against ethanol lesions. The TRH analogue RX-77368 (1.5 ng ic) reduced by 78 and 66% gastric hemorrhagic lesions induced by intragastric intubation of 60 and 80% ethanol, respectively. alpha-CGRP (1 nmol/kg iv) inhibited by 88% gastric lesions induced by 60% ethanol, and this peptide action was blocked by the CGRP antagonist, CGRP-(8-37) (128 nmol/kg iv). The protective effect of RX-77368 against 60% ethanol was completely abolished by the CGRP monoclonal antibody 4901 (4.8 mg/kg iv), CGRP-(8-37) (128 nmol/kg iv), and capsaicin pretreatment (125 mg/kg). Gastric lesions induced by 60% ethanol were not altered by the CGRP antagonist or antibody alone but were enhanced by capsaicin pretreatment. These results suggest that the gastroprotection induced by intracisternal TRH analogue involves an interaction between central vagal efferent pathways and splanchnic sensory afferent terminals containing CGRP.
Administration of 8-OH-DPAT into the midbrain raphe nuclei: effects on medial hypothalamic NE-induced feeding
Authors: Currie P. J. , Fletcher P. J. , Coscina D. V.
Published: December 2017

The 5-hydroxytryptamine (5-HT)1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) has been shown to elicit dose-dependent feeding in satiated rats when injected into the midbrain raphe nuclei. Because the feeding-stimulatory effects of this compound are associated with reduced 5-HT release in forebrain, and because recent evidence suggests that medial hypothalamic norepinephrine (NE) and 5-HT may function antagonistically in the control of feeding, the present study sought to determine whether 8-OH-DPAT would enhance feeding induced by infusing NE into the medial hypothalamus. All experiments were conducted using ad libitum-fed adult male rats with two indwelling cannulas, one aimed at the paraventricular nucleus (PVN), the other at either the dorsal or median raphe. In the first series of experiments rats received a fixed dose of 8-OH-DPAT (0.8 nmol) microinjected into the dorsal or median raphe, as well as NE (0, 10, 20 nmol) administered into the PVN. In the second series, rats received varying doses of raphe 8-OH-DPAT (0, 0.4, 0.8 nmol) coinjected with a single dose of PVN NE (20 nmol). Results indicated that 8-OH-DPAT and NE both elicited reliable increases in 1-h food intake, although their effects did not interact. In the final series, rats were injected with subthreshold doses of both compounds. While combined injections of 8-OH-DPAT and NE modestly increased feeding compared with saline control, this increase was not significantly greater than intake found after injection of 8-OH-DPAT or NE alone.(ABSTRACT TRUNCATED AT 250 WORDS)
Brief mesenteric ischemia increases PGE2, but not PGI2, in intestinal lymph of cats
Authors: Rendig S. V. , Pan H. L. , Longhurst J. C.
Published: December 2017

Mesenteric ischemia of short duration (5-10 min) can stimulate A delta- and C-fiber afferent nerve endings in the viscera to reflexly activate the cardiovascular system. The mechanism of activation of abdominal visceral afferents is probably multifactorial and may involve prostaglandins (PGs), which have been shown to directly stimulate and/or sensitive visceral afferents when administered exogenously. We hypothesized that brief visceral ischemia is accompanied by release of PGI2 and PGE2 into the interstitium, where these cyclooxygenase products could stimulate or sensitize visceral afferent nerve endings. Accordingly, we measured immunoreactive PGE2 (iPGE2) and 6-keto-PGF1 alpha (i6-keto-PGF1 alpha), the stable metabolite of PGI2, in lymph draining the ischemic viscera as well as in portal venous blood. An intestinal lymph duct distal to the lymph node was cannulated in pentobarbital sodium-anesthetized cats. Lymph and plasma iPGE2 and i6-keto-PGF1 alpha concentrations were measured by radioimmunoassay before, during, and immediately after a 5- to 10-min occlusion of the descending aorta. The i6-keto-PGF1 alpha concentration increased significantly (P < 0.001) in portal venous plasma (61 +/- 12 to 107 +/- 18 pg/0.1 ml; n = 14) but not in lymph (148 +/- 30 to 159 +/- 24 pg/0.1 ml; n = 16). In contrast, iPGE2 concentration was significantly (P < 0.01) elevated in both venous plasma (156 +/- 16 to 207 +/- 26 pg/0.1 ml; n = 19) and lymph (520 +/- 48 to 590 +/- 52 pg/0.1 ml; n = 20).(ABSTRACT TRUNCATED AT 250 WORDS)
A novel role for the nitric oxide-cGMP signaling pathway: the control of epithelial function in Drosophila
Authors: Dow J. A. , Maddrell S. H. , Davies S. A. , Skaer N. J. , Kaiser K.
Published: December 2017

The nitric oxide (NO) signaling pathway plays major roles in the vertebrate vascular, nervous, and immune systems. Here we present evidence that all the elements in the NO pathway are present in, and act to control epithelial fluid secretion by, the Malpighian tubules of an insect, Drosophila melanogaster. This finding will allow both a physiological and a molecular genetic dissection of the NO pathway in the same tissue.
1H magnetic resonance imaging of freezing and thawing in freeze-tolerant frogs
Authors: Rubinsky B. , Wong S. T. , Hong J. S. , Gilbert J. , Roos M. , Storey K. B.
Published: December 2017

Proton magnetic resonance imaging (MRI) of the processes of freezing and thawing in the wood frog Rana sylvatica provided noninvasive and real-time analysis of the mode of ice propagation through the body of a freeze-tolerant vertebrate. MRI revealed a directional movement of ice from the exterior inward that required several hours to reach completion. Freezing in core organs such as liver, which produces and exports cryoprotectant, and heart, which circulates it, was delayed and occurred well after the organs were surrounded by extraorgan ice. Natural thawing was a very different process; thawing began uniformly throughout the body, but core organs melted more rapidly than peripheral ones, an adaptation that may be key to the early restoration of heartbeat and breathing. The images presented demonstrate the sensitivity and power of MRI and its potential to become a critical monitoring technology in the development of cryopreservation techniques for mammalian organ explants.
ATP-MgCl2 treatment after trauma-hemorrhage/resuscitation increases hepatocyte P2-purinoceptor binding capacity
Authors: Mahmoud M. S. , Wang P. , Hootman S. R. , Reich S. S. , Chaudry I. H.
Published: December 2017

Although our studies indicate that P2-purinoceptor binding capacity decreases after hemorrhage and resuscitation, it is not known whether ATP-MgCl2 administration after hemorrhage has any beneficial effects on the receptor dynamics. To study this, we performed laparotomy (i.e., trauma induced) on rats and bled them to and maintained them at a mean arterial pressure of 40 mmHg until 40% of maximum bleedout volume was returned in the form of Ringer lactate (RL). The animals were then resuscitated with 3 times the volume of maximum bleedout with RL over 45 min followed by 2 times RL along with ATP-MgCl2 (50 mumol/kg body wt) over 95 min. Hepatocytes were isolated at 4, 17, and 27 h after resuscitation. P2-purinoceptor binding characteristics were determined by using [alpha-35S]ATP. Scatchard analysis revealed high-affinity and low-affinity receptor components in the hepatocytes isolated from sham-operated or hemorrhaged animals with or without ATP-MgCl2 infusion. ATP-MgCl2 ameliorated and subsequently restored the decreased maximum binding capacity (Bmax) of the high-affinity receptor component and significantly improved Bmax of the low-affinity receptor component. ATP-MgCl2 administration also produced a progressive enhancement in the affinity of the low-affinity receptor component. Thus the beneficial effects of ATP-MgCl2 observed after trauma-hemorrhage and resuscitation may be, in part, due to the restoration of P2-purinoceptor binding capacity and the enhancement of the receptor affinity.
Descending pathways of powerful pressor response elicited by suprapontine cerebral ischemia in rabbits
Authors: Horiuchi J. , Takeuchi T.
Published: December 2017

We determined the magnitude of the pressor and sympathoexcitatory responses elicited by suprapontine cerebral ischemia (SCI) and the descending pathways in the rostral medulla mediating them. The suprapontine structures of anesthetized and artificially ventilated rabbits were selectively exposed to cerebral ischemia, by combined occlusions of basilar and common carotid artery. SCI produced a pressor response of 78 +/- 9 (SE) mmHg and an increase in renal sympathetic nerve activity of 289 +/- 21% compared with preischemic levels. The magnitude of the pressor and sympathoexcitatory response to SCI was comparable to those in response to global cerebral ischemia. Microinjection of the neurotoxin kainic acid into the pressor sites of the rostral ventrolateral medulla significantly (P < 0.05) reduced the SCI pressor response to 34 +/- 11% of the prelesion control response. Chemical lesions of the pressor sites in the rostral medial and ventromedial medulla resulted in a significant decrease in the pressor response to SCI to 78 +/- 12% of the control response. These results indicate that the suprapontine structures play an important role in the generation of the powerful pressor response elicited by cerebral ischemia and that the pressor response to SCI is mediated by pressor neurons in the rostral medulla.
Effects of plasma hypernatremia on nucleus tractus solitarius neurons
Authors: Hochstenbach S. L. , Ciriello J.
Published: December 2017

Experiments were done in chloralose-anesthetized, paralyzed, and artificially ventilated rats to investigate the effect of plasma hypernatremia and baroreceptor activation on the excitability of nucleus tractus solitarius (NTS) neurons. Extracellular, single unit recordings were made from 67 histologically verified neurons in the region of NTS. The firing frequency of 29 (43%) NTS neurons was increased by the intracarotid infusion of hypertonic saline (0.5 M). The acute activation of baroreceptors after the intravenous infusion of phenylephrine resulted in the excitation of seven (31.8%) or the inhibition of five (22.7%) of the units responsive to plasma hypernatremia. Thirteen of the hypertonic saline-responsive neurons were recorded in animals in which the organum vasculosum lamina terminalis and subfornical organ were lesioned and/or the left vagus nerve was cut. These data suggest that within NTS there exists a pool of neurons that alter their discharge rate in response to changes in plasma sodium levels and that these neurons may also function in cardiovascular regulation.
REM-sleep timing is controlled homeostatically by accumulation of REM-sleep propensity in non-REM sleep
Authors: Benington J. H. , Heller H. C.
Published: December 2017

Sleep structure in the rat was characterized during uninterrupted full-day recordings using an analytic procedure that identifies rapid eye movement (REM) sleep episodes based on REM-sleep-onset electroencephalograph phenomena, hence independently of REM-sleep duration. The data were used to determine whether REM-sleep timing is controlled homeostatically or by an oscillatory mechanism. The findings and conclusions are that 1) non-REM (NREM) sleep episode duration is positively correlated with prior REM-sleep episode duration, suggesting that REM-sleep expression is permissive of NREM sleep; 2) mean NREM-sleep episode duration decreases after repeated brief REM-sleep episodes (< 30 s), also suggesting that discharge of REM-sleep propensity is essential for NREM-sleep expression; 3) REM-sleep episode duration is independent of prior sleep history, suggesting that REM-sleep maintenance is controlled by factors other than accumulated REM-sleep propensity; 4) brief REM-sleep episodes occur progressively more frequently over the course of the NREM-sleep interval between sustained REM-sleep episodes (> 30 s), suggesting that REM-sleep propensity increases progressively within episodes of NREM sleep; and 5) the diurnal cycle of REM-sleep expression primarily reflects modulation in the efficiency of REM-sleep maintenance. These findings support the hypothesis that REM-sleep timing is controlled by accumulation of REM-sleep propensity during NREM sleep.
beta-Adrenergic control of lipolysis in primate white fat cells: a comparative study with nonprimate mammals
Authors: Bousquet-Melou A. , Galitzky J. , Carpene C. , Lafontan M. , Berlan M.
Published: December 2017

The beta-adrenoceptor subtypes involved in the control of lipolysis in white fat cells of rat, dog, marmoset (Callithrix jacchus), baboon (Papio papio), macaque (Macaca fascicularis), and human were compared. In all species [3H]CGP-12177 binding (up to 3 nM) indicated the existence of a homogeneous population of binding sites in fat cell membranes, and competition studies showed that beta 1- and beta 2-adrenoceptors were present. Selective beta 1 or beta 2-adrenoceptor agonists induced lipolysis. The efficiencies of isoproterenol and norepinephrine were similar. The use of selective beta 3-adrenoceptor agonists revealed that BRL-37344 and CL-316243 were full agonists, whereas CGP-12177 and SR-58611A were partial agonists in rat and dog white fat cells. beta 3-Agonists partially stimulated lipolysis in the marmoset, while CGP-12177 was weakly active in the baboon. In macaque and human fat cells, beta 3-agonists were ineffective. The lipolytic effect of norepinephrine involves beta 1-and/or beta 2-adrenoceptors in baboon, macaque, and human. The baboon and macaque constitute valuable models for studying the beta-adrenergic control of lipolysis.
Characterization and postnatal development of 5'-deiodinase activity in goat perirenal fat
Authors: Nicol F. , Lefranc H. , Arthur J. R. , Trayhurn P.
Published: December 2017

The capacity to generate the active thyroid hormone 3,3',5-triiodothyronine (T3) from thyroxine (T4) has been assessed in the major adipose tissue depot (perirenal) of newborn goats through measurements of iodothyronine 5'-deiodinase activity. High activity of the enzyme was found in the perirenal adipose tissue. Developmental studies demonstrated that there was a rise, then a fall, in iodothyronine 5'-deiodinase activity (per milligram of protein) over the first week of postnatal life, with a further decline between the second and third weeks. At 3 wk of age, the activity per gram of tissue was only 2% of that of the newborn; however, because of age-related increases in the lipid content of the perirenal adipose tissue, the activity per cell (per microgram of DNA) at 3 wk of age was approximately 10% of the activity in the newborn. Kinetic studies and inhibition characteristics with propylthiouracil, gold thioglucose, and T4 indicated that the iodothyronine 5'-deiodinase in goat perirenal adipose tissue is of the type I form; there was no evidence for the type II enzyme. Affinity labeling of the iodothyronine 5'-deiodinase with bromoacetyl-rT3 or bromoacetyl-T4 was consistent with this view. In contrast to goats, only the type II enzyme is evident in rats and mice. It is concluded that the perirenal (brown) adipose tissue is likely to be a significant source of T3 for other tissues in newborn goats and that there are major species variations in the form of iodothyronine 5'-deiodinase present in brown fat, with the goat exclusively exhibiting type I.
SIN-1 reverses attenuation of hypercapnic cerebrovasodilation by nitric oxide synthase inhibitors
Authors: Iadecola C. , Zhang F. , Xu X.
Published: December 2017

We sought to determine whether the attenuation of the hypercapnic cerebrovasodilation associated with inhibition of nitric oxide synthase (NOS) can be reversed by exogenous NO. Rats were anesthetized (halothane) and ventilated. Neocortical cerebral blood flow (CBF) was monitored by a laser-Doppler probe. The NOS inhibitor N omega-nitro-L-arginine methyl ester (L-NAME; 40 mg/kg iv) reduced resting CBF [-36 +/- 5% (SE); P < 0.01, analysis of variance] and attenuated the increase in CBF elicited by hypercapnia (partial pressure of CO2 = 50-60 mmHg) by 66% (P < 0.01). L-NAME reduced forebrain NOS catalytic activity by 64 +/- 3% (n = 10; P < 0.001). After L-NAME, intracarotid infusion of the NO donor 3-morpholinosydnonimine (SIN-1; n = 6) increased resting CBF and reestablished the CBF increase elicited by hypercapnia (P > 0.05 from before L-NAME). Similarly, infusion of the guanosine 3',5'-cyclic monophosphate (cGMP) analogue 8-bromo-cGMP (n = 6) reversed the L-NAME-induced attenuation of the hypercapnic cerebrovasodilation. The NO-independent vasodilator papaverine (n = 6) increased resting CBF but did not reverse the attenuation of the CO2 response. SIN-1 did not affect the attenuation of the CO2 response induced by indomethacin (n = 6). The observation that NO donors reverse the L-NAME-induced attenuation of the CO2 response suggests that a basal level of NO is required for the vasodilation to occur. The findings are consistent with the hypothesis that NO is not the final mediator of smooth muscle relaxation in hypercapnia.(ABSTRACT TRUNCATED AT 250 WORDS)
Glucose uptake and GLUT-4 protein distribution in skeletal muscle of the obese Zucker rat
Authors: Brozinick J. T. , Etgen G. J. , Yaspelkis B. B. , Ivy J. L.
Published: December 2017

The rates of muscle glucose uptake of lean and obese Zucker rats were assessed by hindlimb perfusion under basal conditions (no insulin), in the presence of a maximally stimulating concentration of insulin (10 mU/ml), and after muscle contraction elicited by electrical stimulation of the sciatic nerve. After perfusion, plasma and microsomal membranes were isolated from selected hindlimb muscles for determination of GLUT-4 protein distribution. Under basal conditions, rates of glucose uptake were similar for lean and obese rats despite plasma membranes from lean rats containing 82% more GLUT-4 protein than obese rats. Insulin stimulation resulted in significant increases in plasma membrane GLUT-4 protein concentration in lean but not obese rats. Glucose uptake of lean rats (35.3 +/- 4.7 mumol.h-1.g-1) in the presence of insulin was approximately fourfold greater than that of obese rats (8.8 +/- 1.3 mumol.h-1.g-1), but this difference in glucose uptake could not be completely accounted for by the difference in plasma membrane GLUT-4 protein concentration. Stimulation by contraction resulted in significant increases in plasma membrane GLUT-4 protein concentration in both lean and obese rats and similar rates of glucose uptake. These results suggest that the muscle insulin resistance of the obese Zucker rat is due to 1) a reduced plasma membrane GLUT-4 protein concentration, which results in part from an impairment in the insulin-stimulated GLUT-4 protein translocation process, and 2) a defect in the insulin-stimulated activation of this protein. However, contraction-stimulated glucose uptake, GLUT-4 protein translocation, and activation are normal in the obese Zucker rat.
Fractal nature of short-term systolic BP and HR variability during lower body negative pressure
Authors: Butler G. C. , Yamamoto Y. , Hughson R. L.
Published: December 2017

We have shown previously that the heart rate variability (HRV) signal is fractal in nature with a high degree of complexity, as given by the calculated fractal dimension (DF). We have also reported that loss of complexity, as indicated by a reduction in DF of HRV, is associated with orthostatic hypotension and impending syncope. To extend this investigation of cardiovascular responses, we have investigated the signal characteristics of short-term systolic blood pressure variability (BPV) coincident with measurements of HRV during orthostatic stress. Eight healthy men completed a test protocol of 20 min supine rest followed sequentially by 10 min at each of -5, -15, -25, -40, and -50 mmHg lower body negative pressure (LBNP) and 10 min supine recovery. We found that resting BPV and HRV were fractal with approximately 70% of both variables in the fractal component of the variability signal. The slope of the 1/f beta relationship was 1.16 +/- 0.12 for HRV and 2.31 +/- 0.17 for BPV. With increasing levels of orthostatic stress, the 1/f beta slope of HRV increased significantly to 1.68 +/- 0.08 at -50 mmHg LBNP, whereas the 1/f beta slope was unchanged for BPV. Indicators of parasympathetic and sympathetic nervous system activity derived from heart rate variability suggested reduced and increased values, respectively, as the LBNP increased. These data indicate important differences in heart rate and blood pressure control under orthostatic stress.
Pharmacological dissociation of responses to CCK and gastric loads in rat mechanosensitive vagal afferents
Authors: Schwartz G. J. , McHugh P. R. , Moran T. H.
Published: December 2017

To identify the transduction mechanisms underlying gastric vagal afferent responses to gastric loads and cholecystokinin (CCK), we investigated the ability of specific CCK antagonists, acute pylorectomy, and cholinergic blockade to effect these vagal afferent responses. The CCK-B antagonist L-365,260 (10 pmol-1 nmol) failed to block the gastric vagal afferent response to gastric loads or 100 pmol CCK, while the CCK-A antagonist devazepide (100 pmol-100 nmol) competitively and dose dependently attenuated the response to CCK but not to gastric loads. Application of 100 nmol of the low-affinity CCK receptor antagonist CCK-JMV-180 also completely blocked the gastric vagal afferent response to 100 pmol CCK. Acute pylorectomy failed to block the gastric vagal afferent response to 100 pmol CCK or 2-ml gastric loads. Atropine sulfate administration (15 mg/rat) failed to block the gastric vagal afferent response to 100 pmol CCK or 2-ml gastric loads. These data suggest that 1) the vagal afferent response to CCK is mediated through CCK's interactions with vagal, rather than pyloric, CCK-A receptors, and 2) the vagal afferent responses to CCK and to gastric loads are mediated by dissociable, possibly independent, transduction mechanisms.