bluuebunny/crossref_metadata_2025 huggingface dataset:
Deoxythymidine kinase activity of human implanted sponge connective tissue in zinc deficiency.
Authors: Prasad A S , Fernandez-Madrid F , Ryan J R
Published: December 2017

The activity of deoxythymidine kinase was assayed in implanted sponge connective tissue in three groups of subjects: 1) five normal controls (having normal levels of plasma and red cell zinc); 2) four patients with sickle cell anemia who had low zinc in red cells and hair; and 3) two volunteers (under strict dietary controls), after 6 mo of zinc restriction (2.7 mg/day) and repeated after 3 mo of zinc repletion (30 mg/day). Total protein, total collagen, RNA/DNA, and deoxythymidine kinase activity were measured by techniques reported previously. In sickle cell anemia patients, deoxythymidine kinase activity was not detected, and RNA/DNA, total collagen, and total protein contents were decreased compared to normal controls (statistically significant). In human volunteers deoxythymidine kinase activity was not detected during the zinc restriction phase. After supplementation with zinc, deoxythymidine kinase activity became 70% of normal control levels, and RNA/DNA, total collagen, and total protein contents of sponge connective tissue increased. In conclusion, an adverse effect of zinc deficiency on deoxythymidine kinase activity of implanted sponge connective tissue of man has been demonstrated for the first time.
Kidney metabolite levels and ammonia production in acute acid-base alterations in the rat.
Authors: Boyd T A , Goldstein L
Published: December 2017

Acute acid-base disturbances cause rapid changes in renal ammonia production in the rat. To investigate the regulation of ammonia production in acute acid-base stresses, we examined the effects of such conditions on tissue levels of metabolites such as alpha-ketoglutarate (KG), which inhibits kidney mitochondrial glutamine uptake and deamidation at physiological concentrations. In rats given low (5 mmol/kg) or high (20 mmol/kg) doses of NH4Cl by stomach tube or placed in a 10% CO2 atmosphere, kidney KG levels fell after 1-h by 44, 69, or 73%, respectively. NaHCO3 administration produced no change in KG levels. Renal glutamate and glutamine levels changed little, if at all, in any group, and renal phosphoenolpyruvate levels were not altered except for a decrease in the NH4HCO3 group. In livers of the same rats, treatments produced different patterns of metabolite changes; KG fell only in the NaHCO3 and NH4HCO3 groups. These results support the concept that KG is a specific regulator of renal ammonia production in acute acid-base stresses.
Intraventricular 2-deoxyglucose, glucose, insulin, and free fatty acid mobilization.
Authors: Coimbra C C , Gross J L , Migliorini R H
Published: December 2017

The possibility of eliciting changes in the activity of lipomobilizing centers in the rat central nervous system was investigated by injecting glucose or substances that affect glucose metabolism directly into the cerebrospinal fluid of conscious, unrestrained rats. Intracerebroventricular administration of minute amounts of 2-deoxyglucose to fed rats induced a rapid increase in the concentration of plasma free fatty acids (FFA) without affecting plasma insulin levels. Furthermore, small amounts of either glucose or insulin injected intraventricularly reduced the increased plasma FFA levels of normal fasted rats without affecting the glycemia or plasma insulin levels in these animals. In diabetic rats, the increased levels of plasma FFA were not affected by intraventricular administration of glucose, but clearly decreased after intraventricular insulin. The results suggest that the lipomobilizing centers are insulin sensitive and that their activity is modulated by the rate of glucose uptake, rather than by external hexose concentration. The data are also consistent with our previous hypothesis of a direct control of the sympathetic tonus of adipose tissue by these centers.
Cation-anion gap and choleretic properties of rat bile.
Authors: Klos C , Paumgartner G , Reichen J
Published: December 2017

To investigate whether bile contains choleretic anion(s) other than bile salts (BS) that could account for the observed cation-anion gap in bile, the choleretic properties of bile were investigated in the rat. Infusion of bile increased bile flow significantly more than did taurocholate (TC) (P less than 0.005). By contrast, TC increased BS excretion more substantially (P less than 0.01). This effect was dose dependent for both bile and TC. The choleretic principle had a molecular weight of less than 1,000 as estimated by ultrafiltration of bile. Infusion of bile that had been chromatographed on BioBeads SM-2 still elicited choleresis, whereas bile that had been chromatographed on Dowex 1 x 50W did not. Administration of bile in vivo did not affect Na+-K+-stimulated ATPase activity in liver plasma membrane fractions. These results suggest that bile contains anion(s) other than 3-hydroxy-BS, which increase bile flow in a dose-dependent fashion without affecting the permeability of the biliary tree. This putative choleretic appears to be anionic in nature, heat stable, and has an apparent molecular weight of less than 1,000. This finding suggested that bile salt-independent bile flow partly depends on the excretion of a currently undefined anion.
Intestinal absorption of 25-hydroxyvitamin D3 in unanesthetized rat.
Authors: Hollander D , Rim E , Morgan D
Published: December 2017

We investigated the mechanism and characteristics of 25-hydroxyvitamin D3 (25-OH-D3) absorption in the unanesthetized rat by using a single-pass intestinal perfusion technique. The rate of 25-OH-D3 absorption remained linear for a wide range of concentrations (2-900 nM). Absorption rate of 25-OH-D3 increased as the pH, the bile acid concentration, and thickness of the unstirred water layer were decreased. Absorption did not change after the additions of fatty acids of varied chain lengths and degrees of saturation. In rats with lymph and bile fistulas, 18.5% and 16.3% of the infused radio-activity appeared in the lymph and bile drainage, respectively. These experiments indicate that 25-OH-D3 is absorbed by a passive diffusion mechanism that is influenced by the intestinal pH, bile acid concentration, and thickness of the unstirred water layer, but not by the presence of fatty acids. Approximately equal fractions of the infused hydroxylated vitamin are recovered from the lymphatic and biliary fluids.
Peristalsis in distal colon of the rabbit: an analysis of mechanical events.
Authors: Pescatori M , Grassetti F , Ronzoni G , Mancinelli R , Bertuzzi A , Salinari S
Published: December 2017

The mechanical behavior of isolated segments of rabbit distal colon was studied by experimental and theoretical techniques. The isometric isovolumic preparation was adopted to obtain records of endoluminal pressure, force, and wall morphology during 59 spontaneous and 53 electrostimulated peristaltic contractions. On strips of longitudinal and circular muscle, excised from the same segments, the tension-velocity and tension-length relationships were also measured. To describe mechanical events occurring during propulsion, a mathematical model was developed that incorporates the measured characteristics of the muscle. In response to given waveforms for the activation of the two muscular coats, the solution of model equations provides the time courses of pressure, force, and wall morphology. A number of parameters measured in the experimental contractions (e.g., pressure and force peaks, time lag between the stimulus and the peaks, velocity of progression of the peristaltic wave) were compared with model outputs, and a similar pattern of response was found. Therefore the model made it possible to relate segment and strip experimental data and to investigate the temporal and quantitative relationship between the contraction of longitudinal and circular layer.
Anomalous calcium secretion in rat ileum: role of paracellular pathway.
Authors: Nellans H N , Kimberg D V
Published: December 2017

The mechanism of apparent calcium secretion by unstripped rat ileum in vitro has been investigated using mannitol and polyethylene glycol (mol wt 900) as markers for extracellular transepithelial flux. In the absence of electrochemical gradients between the mucosal and serosal bathing media in a modified Ussing chamber, net fluxes of both mannitol and polyethylene glycol were observed from serosa to mucosa in the presence of 11 mM D-glucose. Mucosal-to-serosal calcium flux reveals a significant cellular component, but serosal-to-mucosal calcium permeability is a linear function of mannitol permeability, suggesting an exclusively extracellular route. For the mucosal-to-serosal fluxes, inhibition of calcium flux by 1 mM N-ethylmaleimide results in a calcium-to-mannitol permeability ratio indistinguishable from that measured for serosal-to-mucosal flux. This evidence suggests that the apparent calcium secretion observed at 10 mM medium calcium is not the result of a cellular secretory mechanism. It is proposed that a hydrostatic driving force generated internal to the tissue but external to the cells results in net calcium secretion at calcium conentrations that saturate the cellular absorptive mechanism.
Mechanism of inhibition of alanine absorption by Na ricinoleate.
Authors: Hajjar J J , Murphy D M , Scheig R L
Published: December 2017

The mechanism of inhibition of alanine absorption by Na ricinoleate has been examined in the rabbit intestine. This fatty acid in a concentration of 2--5 mM inhibits alanine absorption in vivo and in vitro. The inhibition is more evident in the jejunum than in the ileum. Strips of ileal mucosa treated with Na ricinoleate gain Na. Sodium ricinoleate inhibits alanine influx across rabbit ileum, even in the presence of a sodium gradient across these cells. The results suggest that the main action of Na ricinoleate is on the alanine-transport system at the brush-border membrane. The fatty acid may also inhibit amino acid absorption by increasing intestinal cell Na concentration, which results in a decreased Na gradient across the brush-border membrane.
Calcium absorption and intestinal calcium-binding protein: quantitative relationship.
Authors: Feher J J , Wasserman R H
Published: December 2017

The concentration of the vitamin D-induced calcium-binding protein (CaBP) and calcium absorption from the duodenum were investigated in chicks with an in vivo ligated-loop technique. The relation between CaBP and calcium absorption was dependent on a) source of vitamin D activity (either vitamin D3 or 1,25-dihydroxycholecalciferol); b) dosage of vitamin D3; c) time after administration of vitamin D3 to rachitic animals. To aid in the interpretation of these results, a phenomenological model was developed in which CaBP was viewed as being linearly related to a portion of calcium absorption. The model, when applied to the data, suggests that there is a "nonfunctional" pool of CaBP the size of which is determined by the vitamin D status of the animal. After correction for this nonfunctional pool, the proportionality between CaBP and calcium absorption is independent of the vitamin D status of the animal.
Relationship of luteal blood flow and corpus luteum function in pseudopregnant rats.
Authors: Pang C Y , Behrman H R
Published: December 2017

An experiment was conducted to examine whether blood flow to corpora lutea may regulate luteal function as judged from plasma levels of progesterone (P) in mature pseudopregnant rats. 141Ce-labeled microspheres (14.1 +/- 0.8 micrometer diam) were used to measure cardiac output and organ and tissue blood flow in rats on days 6, 8, 10, and 12 of pseudopregnancy and in proestrus following luteal regression. The mean arterial blood pressure and cardiac output were similar among all groups of rats. Although a significant (P less than 0.05) decrease in plasma P was observed in rats on day 12 of pseudopregnancy, no change in luteal blood flow or distribution of ovarian blood flow to the corpora lutea was seen at this stage of pseudopregnancy when compared to day 8 or 10 of pseudopregnancy. However, a significant decrease (P less than 0.05) in luteal blood flow was seen in proestrous rats. Because a decrease in plasma progesterone preceded the decrease in luteal blood flow, it was concluded that physiological luteal regression may not be initiated by a reduction of blood flow to the corpus luteum.
Effect of ethanol on disaccharidases of hamster jejunal brush border membrane.
Authors: Dinda P K , Hurst R O , Beck I T
Published: December 2017

This study was undertaken to investigate the effect of alcohol on the activity of jejunal disaccharidases (DS). The activity of DS in a preparation of purified brush border membrane of hamster jejunum was measured in the absence and in the presence (0.8 to 6.4% wt/vol) of ethanol. To compare the effect of alcohol on DS with its action on a brush border enzyme of a different group, we also measured the activity of alkaline phosphatase (AP) under similar conditions. Ethanol depressed the activity of sucrase, maltase, and lactase in a dose-dependent and time-dependent manner, but it stimulated the activity of AP. The ethanol-induced inhibition of DS was completely reversible. Kinetic studies indicate that ethanol depressed the Vmax and increased the Km of sucrase and lactase. The Vmax of maltase also decreased, but the Km of this hydrolase was not affected by ethanol. From the results of this study it would appear that acute exposure of the jejunal brush border to ethanol depresses the DS activity of the membrane and that (because the AP was not depressed) the ethanol-induced inhibition of DS is not the result of a general inhibition of all enzymes of the brush border.
Changes in gastrin levels, food intake, and duodenal mucosal growth during lactation.
Authors: Lichtenberger L M , Trier J S
Published: December 2017

Gastrointestinal epithelial cell proliferation is stimulated during lactation in the rat. Serum gastrin levels and food intake are also increased during lactation. We investigated the role of gastrin and food intake as possible mediators of duodenal mucosal growth during the first 15 days of lactation. As the lactation period progressed, significant increases in the following crypt properties were noted: 1) crypt length; 2) cells/crypt; 3) labeling index; and 4) dimensions of the proliferative zone. Maternal serum gastrin levels rose abruptly by the first day of birth and remained elevated throughout lactation. The increases in crypt cell proliferation significantly correlated with food intake but not with serum gastrin levels during lactation. Mucosal mass and villus-crypt dimensions were also significantly increased above virgin levels in lactating antrectomized rats. These results suggest that the increase in duodenal growth during lactation most probably is not mediated by postpartum hypergastrinemia and that the increase in cell proliferation may be a direct response to an enhancement in food intake.
Glucose clamp technique: a method for quantifying insulin secretion and resistance.
Authors: DeFronzo R A , Tobin J D , Andres R
Published: December 2017

Methods for the quantification of beta-cell sensitivity to glucose (hyperglycemic clamp technique) and of tissue sensitivity to insulin (euglycemic insulin clamp technique) are described. Hyperglycemic clamp technique. The plasma glucose concentration is acutely raised to 125 mg/dl above basal levels by a priming infusion of glucose. The desired hyperglycemic plateau is subsequently maintained by adjustment of a variable glucose infusion, based on the negative feedback principle. Because the plasma glucose concentration is held constant, the glucose infusion rate is an index of glucose metabolism. Under these conditions of constant hyperglycemia, the plasma insulin response is biphasic with an early burst of insulin release during the first 6 min followed by a gradually progressive increase in plasma insulin concentration. Euglycemic insulin clamp technique. The plasma insulin concentration is acutely raised and maintained at approximately 100 muU/ml by a prime-continuous infusion of insulin. The plasma glucose concentration is held constant at basal levels by a variable glucose infusion using the negative feedback principle. Under these steady-state conditions of euglycemia, the glucose infusion rate equals glucose uptake by all the tissues in the body and is therefore a measure of tissue sensitivity to exogenous insulin.
Effect of winter sleep on pituitary-thyroid axis in American black bear.
Authors: Azizi F , Mannix J E , Howard D , Nelson R A
Published: December 2017

During winter sleep the black bear has decreased levels of serum total and free thyroxine (T4) and triiodothyronine (T3) and a prolonged, delayed response of serum thyrotropin (TSH) (bioassay) to thyrotropin-releasing hormone (TRH). Four weeks after the end of winter sleep, levels of serum thyroid hormones increase, and TSH response to TRH is short and brisk. Serum T4 and T3 rise after TRH administration both during and after winter sleep; however, the maximum increment in serum T3 is greater during winter sleep when the TSH rise is also prolonged and exaggerated. These observations suggest that transient hypothyroidism of possible hypothalamic origin occurs in bears during winter sleep.
Gastric pacemaker rhythm in conscious dog.
Authors: Smout A J , van der Schee E J , Grashuis J L
Published: December 2017

The pacemaker rhythm in the stomach of six healthy conscious dogs was studied by means of the measurement of sequential electrical control activity (ECA) intervals. Only the rhythm originating in the normal pacemake area was studied. An electrical response activity (ERA) score was used to assess contractile activity. Whereas substantial shortening of nonectopic ECA intervals did not occur, contraction-related interval lengthening was a characteristic phenomenon. The temporal relations between ERA scores and interval durations were found to be dependent on the gastric level at which the signals were derived. The activity front of the interdigestive myoelectric complex (IDMEC) appeared to be correlated with the periodic occurrence of considerably prolonged intervals; this sign could be used for recognition of the IDMEC. The motor quiescence phase of the IDMEC was correlated with small interval-to-interval variation, as was the early postprandial phase.
Mucosal gastrin receptor. II. Physical characteristics of binding.
Authors: Takeuchi K , Speir G R , Johnson L R
Published: December 2017

Gastrin binding to rat gastric mucosal membrane preparations revealed a linear Scatchard plot, suggesting the existence of a single class of binding sites. Further studies revealed the absence of a positive cooperative effect. Because the system appeared to satisfy the conditions for Michaelis-Menten type kinetics, the determination of a Kd of approximately 3.6 X 10(-10) M was justified. These studies also demonstrated the efficacy of the specific activity of our iodinated gastrin. Trypsin treatment abolished specific binding of gastrin to the membrane preparation, indicating the protein nature of the receptor. The importance of maintaining extremely low temperatures during the incubation and wash period is demonstrated by the decrease in both the rate and the amount of dissociation at 0 degrees C compared with 30 degrees C.
Model for mucosal circulation of rabbit small intestine.
Authors: Levitt D G , Sircar B , Lifson N , Lender E J
Published: December 2017

A theoretical model of the vascular network of normal rabbit small intestine mucosa was constructed from tissues injected with silicone latex. The segmental resistances of the network were calculated from Poiseuille's law by use of lengths and radii measured microscopically and a value of 0.03 P for the viscosity of blood. Where required for the theoretical calculations, published values were used for pressures in arteries down to 20 micrometer in diameter. Values for total-organ blood flow, its distribution between mucosa and nonmucosa, and the partition of mucosal flow between crypts and villi were similar to values measured in the same animals with microspheres. Blood pressures in arteries smaller than 20 micrometer in diameter were also similar to published values. Mean mucosal capillary pressures were close to published experimental estimations. The model seems to provide a useful starting point for describing the magnitudes and patterns of pressures and flow in the small intestine and for consideration of specific location of the sites of their control.
Effects of osmotic gradients on water and solute transport: in vivo studies in rat duodenum and ileum.
Authors: Miller D L , Hamburger S A , Schedl H P
Published: December 2017

We examined effects of luminal osmolality on net water and solute movements in rat duodenum and ileum. Solutions of sodium chloride (permeating solute) or mannitol (nonpermeating solute) at hypo-, iso-, or hyperosmotic concentrations were recirculated through in situ segments. Luminal osmolality increased towards that of plasma with hyposmotic solutions of both solutes. With isosmotic solutions, luminal osmolality did not change with sodium chloride, but increased with mannitol. With hyperosmotic solutions, luminal osmolality always decreased toward that of plasma with sodium chloride; with mannitol, however, decreases were significant only when initial concentrations were above 400 mosmol/kg. The decrease in osmolality of hyperosmotic sodium chloride resulted from sodium absorption and water secretion. Thus, both hypo- and hyperosmotic solutions of sodium chloride adjusted toward isomolality with plasma by the usual mechanisms of water and solute movement. With mannitol, however, osmotic adjustment of hypertonic luminal contents was restricted or even absent due to movement of sodium down its concentration gradient and reduced hydraulic conductivity of the gut.
Effect of insulin-induced hypoglycemia on protein metabolism in vivo.
Authors: Hourani H , Williams P , Morris J A , May M E , Abumrad N N
Published: December 2017

The effects of insulin-induced hypoglycemia (IIH) on leucine kinetics (mumol.kg-1.min-1) and interorgan flow of amino acids (AA) were examined in 2 groups of 18-h fasted conscious dogs. Insulin was infused at 5 mU.kg-1.min-1 for 3 h. IIH (40 +/- 5 mg/dl) resulted in a drop in plasma leucine (114 +/- 10 to 64 +/- 9 microM) and leucine rate of appearance (Ra) (3.1 +/- 0.1 to 2.4 +/- 0.2) within 1 h but gradually increased (P less than 0.05) to 145 +/- 30 microM and 3.8 +/- 0.5 by 3 h. Leucine oxidative rate of disposal (Rd) increased from 0.44 +/- 0.08 to 1.02 +/- 0.35 (P less than 0.01), and nonoxidative Rd dropped initially but was near basal levels by 3 h. When euglycemia was maintained, there was sustained drop in plasma leucine from 122 +/- 12 to 42 +/- 6 mumol/l, leucine Ra from 3.1 +/- 0.4 to 1.8 +/- 0.2, oxidative Rd from 0.36 +/- 0.03 to 0.22 +/- 0.04, and nonoxidative Rd from 2.75 +/- 0.4 to 1.6 +/- 0.2 (all P less than 0.01). IIH was associated with a significant net release of leucine (and other AA) across the gut (0.04 +/- 0.05 to 1.86 +/- 0.30 mumol.kg-1.min-1; P less than 0.05). In the group with euglycemia there was no significant change in the gut balance of leucine. We conclude that IIH is associated with a proteolytic response and that the gut is the major contributor to this response.
Glucose metabolism and recycling by hepatocytes of OB/OB and ob/ob mice.
Authors: Lahtela J T , Wals P A , Katz J
Published: December 2017

Hepatocytes were prepared from livers of ob/ob (obese diabetic) mice and their lean (OB/OB) siblings that had been fasted for 24 h. The hepatocytes were incubated with [U-14C, 2-3H]-, [U-14C, 3-3H]-, and [U-14C, 6-3H]glucose at concentrations from 20 to 120 mM. 14C was recovered mainly in CO2, glycogen, and lactate. Tritium was recovered in water and glycogen. The yield in labeled products from [2-3H]glucose ranged from two to three times that from [U-14C]glucose. The yields from [3-3H]- and [6-3H]glucose were similar, and 1.3-1.7 times that from [U-14C]glucose. At 40 mM, total utilization of glucose by obese mice was about twice that for lean mice, but there was little difference at 120 mM. The rate of recycling between glucose and glucose 6-phosphate was calculated. An equation to calculate the rate of recycling of glucose from the 2-3H/U-14C ratio in glycogen is derived in the APPENDIX. Our results show that 1) the utilization of glucose by hepatocytes from obese diabetic mice exceeds that of their lean controls, 2) the rate of glucose phosphorylation in both groups greatly exceeds glucose uptake and the rate of glycogen synthesis, 3) glucose phosphorylation represents a difference between a high glucokinase rate and hydrolysis of glucose 6-phosphate, and 4) recycling of glucose carbon between glucose 6-phosphate and pyruvate occurs within mouse hepatocytes.
Insulin stimulates mitochondrial protein synthesis and respiration in isolated perfused rat heart.
Authors: McKee E E , Grier B L
Published: December 2017

The rates of synthesis of mitochondrial proteins by both the cytoplasmic and mitochondrial protein synthetic systems, as well as parameters of respiration, were measured and compared in mitochondria isolated from fresh, control perfused, and insulin-perfused rat hearts. The respiratory control ratio (RCR) in mitochondria from fresh hearts was 8.1 +/- 0.4 and decreased to 6.0 +/- 0.2 (P less than 0.001 vs. fresh) in mitochondria from control perfused hearts and to 6.7 +/- 0.2 (P less than 0.005 vs. fresh and P less than 0.02 vs. control perfused) for mitochondria from hearts perfused in the presence of insulin. A positive correlation between the RCR and the rate of mitochondrial translation was demonstrated in mitochondria from fresh hearts. In mitochondria isolated from control perfused hearts, the rate of protein synthesis decreased to 84 +/- 3% of the fresh rate after 30 min of perfusion and fell further to 64 +/- 3% after 3 h of perfusion. The inclusion of insulin in the perfusion buffer stimulated mitochondrial protein synthesis 1.2-fold by 1 h (P less than 0.005) and 1.34-fold by 3 h of perfusion (P less than 0.001). The addition of insulin to 1-h control perfused hearts shifted the rate of mitochondrial protein synthesis from the control level to the insulin-perfused level within 30 min of additional perfusion, whereas 1 h was required to shift the RCR values of these mitochondria from control levels to insulin-perfused levels. Thus, whereas RCR was a useful predictor of mitochondrial translation rates, it did not account for the effects of insulin on mitochondrial translation.(ABSTRACT TRUNCATED AT 250 WORDS)
Effects of naloxone on renin and pressor responses to acute renal hypotension in rats.
Authors: Weaver C J , Johnson M D
Published: December 2017

Reduction of renal perfusion is followed by increases in plasma renin activity (PRA) and arterial pressure. The present experiments were designed to determine if an opiate antagonist would alter pressor or renin responses to acute reduction of renal arterial pressure (RAP) in anesthetized rats. Male Sprague-Dawley rats were anesthetized with Inactin, and an adjustable constrictor device was placed around the abdominal aorta proximal to the renal arteries. One-half of the animals were pretreated with the opiate antagonist naloxone (2 mg/kg iv), and the other one-half were pretreated with saline vehicle. The abdominal aorta was then constricted to reduce RAP by 25% (measured as femoral arterial pressure) in one-half of the animals in each pretreatment group. Compared with vehicle pretreatment, naloxone pretreatment did not alter the PRA response to aortic constriction; however, naloxone did attenuate the pressor response. We conclude that 1) the PRA response to acute reduction of renal arterial pressure is not dependent on an opiate mechanism in the rat, and 2) attenuation of the pressor response to aortic constriction by naloxone in intact rats is not secondary to a suppression of the PRA response.
Plasma membrane domain localization and transcytosis of the glucagon-induced hepatic system A carrier
Authors: Cariappa R. , Kilberg M. S.
Published: December 2017

We have utilized canalicular (cLPM) and basolateral (blLPM) liver plasma membrane vesicles to investigate the domain localization of several Na(+)-dependent amino acid transporters. Neutral amino acid transport by systems N and ASC was measurable in both domains but was greater in blLPM vesicles. Sodium-dependent glutamate uptake (system X-) was preferentially localized to cLPM. The absolute activity and domain distribution of these three carriers remained unchanged after treatment of rats with a combination of glucagon and dexamethasone. A low level of basal system A activity was present in both the blLPM and cLPM. Glucagon-induced system A activity was first observed in blLPM vesicles approximately 60 min posthormone treatment and, in cLPM vesicles, approximately 30 min later. In situ perfusion of glucagon-treated rat liver with the membrane-impermeant protein modification reagent glutathione maleimide specifically inactivated blLPM system A activity and abolished the delayed arrival of hormone-induced activity to the cLPM. Transient perfusion of the liver with glutathione maleimide followed by a recovery period in vivo showed that the reagent did not irreversibly inactivate the transcytotic process and also provided an independent demonstration of the time delay between arrival of the carrier activity at the two membrane surfaces. These results support the concept of a transcytotic process in which the blLPM is the sorting site for the hormone-induced system A carrier proteins that eventually reach the cLPM.
1,25(OH)2D3 blunts hormone-elevated cytosolic Ca2+ in osteoblast-like cells
Authors: Green J. , Kleeman C. R. , Schotland S. , Ye L. H.
Published: December 2017

Cytosolic free calcium ([Ca2+]i) is an important regulator of bone cell physiology. We studied the interaction of vitamin D metabolites on the hormonal-activated Ca message system in the osteoblastic cell line UMR-106. The acute rise in [Ca2+]i induced by different calciotropic hormones [parathyroid hormone, prostaglandin E2 (PGE2)] was dose dependently blunted by 1,25-dihydroxyvitamin D [1,25(OH)2D3; half-maximal inhibitory concn approximately 5 x 10(-11) M] and was initially observed after 8 h of preincubation. The 1,25(OH)2D3 metabolite of vitamin D was two orders of magnitude more potent than 24,25(OH)2D3 and 25(OH)D3. To discern between an effect of 1,25(OH)2D3 on hormonal-induced Ca2+ entry through the plasma membrane channel vs. release of Ca2+ from internal stores, we suspended fura-2-loaded cells in Mn2+ rather than Ca2+ buffers. In cells preincubated with 1,25(OH)2D3, [Ca2+]i release (indicated by [Ca2+]i transient) was significantly blunted, whereas Mn2+ influx (indicating Ca2+ flux across the plasma membrane) was unaltered, suggesting a selective effect of 1,25(OH)2D3 on hormonal-activated release of Ca2+ from intracellular stores. 1,25(OH)2D3 also inhibited the PGE2-induced production of inositol 1,4,5-trisphosphate. We conclude that, in osteoblasts, chronic (hours) incubation with 1,25(OH)2D3 leads to attenuated stimulation of the [Ca2+]i transduction pathway by calciotropic hormones. This effect of 1,25(OH)2D3 may provide a cellular basis for the synergism between the effects of vitamin D and calciotropic hormones at the bone level.
Hypoxia causes glycogenolysis without an increase in percent phosphorylase a in rat skeletal muscle
Authors: Ren J. M. , Gulve E. A. , Cartee G. D. , Holloszy J. O.
Published: December 2017

Stimulation of skeletal muscle to contract activates phosphorylase b-to-a conversion and glycogenolysis. Despite reversal of the increase in percentage of phosphorylase a after a few minutes, continued glycogen breakdown can occur during strenuous exercise. Hypoxia causes sustained glycogenolysis in skeletal muscle without an increase in percentage of phosphorylase a. We used this model to obtain insights regarding how glycogenolysis is mediated in the absence of an increase in percentage of phosphorylase a. Hypoxia caused a 70% decrease in glycogen in epitrochlearis muscles during an 80-min incubation despite no increase in percentage of phosphorylase a above the basal level of approximately 10%. Muscle Pi concentration increased from 3.8 to 8.6 mumol/g muscle after 5 min and 15.7 mumol/g after 20 min. AMP concentration doubled, attaining a steady state of 0.23 mumol/g in 5 min. Incubation of oxygenated muscles with 0.1 microM epinephrine induced an approximately sixfold increase in percentage of phosphorylase a but resulted in minimal glycogenolysis. Muscle Pi concentration was not altered by epinephrine. Despite no increase in percentage of phosphorylase a, hypoxia resulted in a fivefold greater depletion of glycogen over 20 min than did epinephrine. To evaluate the role of phosphorylase b, muscles were loaded with 2-deoxyglucose 6-phosphate, which inhibits phosphorylase b. The rate of glycogenolysis during 60 min of hypoxia was reduced by only approximately 14% in 2-deoxyglucose 6-phosphate-loaded muscles.(ABSTRACT TRUNCATED AT 250 WORDS)
Development and application of a radioimmunoassay to detect interleukin-1 in rat peripheral circulation
Authors: Derijk R. , Berkenbosch F.
Published: December 2017

We describe the development and application of a radioimmunoassay to detect circulating interleukin (IL)-1 beta concentrations in the rat. No IL-1 immunoreactivity above the detection limit of the assay (100 pg/ml) could be detected in plasma of control rats. In contrast, immunoreactive IL-1 was detected after intravenous administration of rat recombinant IL-1 beta (rrIL-1 beta) or bacterial lipopolysaccharide (LPS) to rats. The effect of LPS on plasma immunoreactive IL-1 concentrations was time and dose dependent. The immunoreactive IL-1 response to LPS was prevented by in vivo macrophage depletion induced by liposome-directed macrophage suicide technique. Gel filtration of plasma from LPS-treated rats revealed the presence of a high and a smaller molecular form of immunoreactive IL-1. The small molecular immunoreactive IL-1 peak coeluted with rrIL-1 beta and probably represents the 17-kDa form of IL-1 beta. In conclusion, our data support the hypothesis that IL-1 secreted by macrophages can act as a humoral signal molecule to induce the immunological, metabolic, and neuroendocrine changes in response to bacterial LPS.
Effects of insulin on total RNA, poly(A)+ RNA, and mRNA in primary cultures of rat hepatocytes
Authors: Hsu C. J. , Kimball S. R. , Antonetti D. A. , Jefferson L. S.
Published: December 2017

The purpose of this study was to examine mechanisms involved in the regulation of protein synthesis in primary cultures of rat hepatocytes. Hepatocytes were maintained in a chemically defined serum-free medium in the presence or absence of insulin. The rate of protein synthesis in hepatocytes deprived of insulin between days 2 and 5 of culture was reduced to 67% of the rate observed in insulin-maintained controls. The decrease in protein synthetic rate was accompanied by a proportional fall in the content of both total RNA and poly(A)+RNA, suggesting that the capacity for protein synthesis was reduced in the absence of insulin. Both total RNA and poly(A)+ RNA contents and the protein synthetic rate were returned to control values after 3 days of insulin resupplementation. In addition, the effect of insulin on the expression of specific mRNAs was assessed by in vitro translation of total RNA followed by two-dimensional gel analysis of radiolabeled translation products. Only 13 of the greater than 150 spots discernible on the two-dimensional gels were altered in response to insulin. The mRNAs that were altered include examples of repression and stimulation of expression in response to insulin deprivation. Thus, in isolated rat hepatocytes, insulin regulates the capacity of both overall protein synthesis as well as the capacity for the synthesis of specific proteins.
Effect of gender, body composition, and equilibration time on the 2H-to-18O dilution space ratio
Authors: Goran M. I. , Poehlman E. T. , Nair K. S. , Danforth E.
Published: December 2017

Physiological sources of variation in the 2H-to-18O dilution space ratio (DSR) were examined in 34 males and 20 females (4-78 yr; 14.7-143.2 kg; 1.8-61.0% body fat). Dilution spaces were obtained by time 0 extrapolation of isotope washout over 10-14 days, and body composition was obtained by underwater weight (adults) or bioelectrical impedance (children). The mean DSR was 1.050 +/- 0.015 (range 1.029-1.111), significantly higher (P < 0.001) than the traditionally assumed value of 1.029 based on exchange over 4 h. Use of the value 1.029 causes a systematic 8% overestimate of energy expenditure from doubly labeled water, relative to use of the value 1.05. The DSR was not related to body composition or age but was significantly higher (P < 0.05) in males (1.052 +/- 0.016) than in females (1.044 +/- 0.012). This gender effect was not explained by differences in the number of exchangeable hydrogens in the body. We conclude 1) variation in the 2H-to-18O DSR is not explained by body composition but is influenced by the chemical availability of exchangeable hydrogens to undergo exchange; 2) because the DSR is not easily predicted, use of the observed dilution spaces are recommended; 3) if a fixed DSR is used, values of 1.044 and 1.052 are recommended in females and males, respectively.
Effects of acute hypoxemia on insulin-like growth factors and their binding proteins in fetal sheep
Authors: Iwamoto H. S. , Murray M. A. , Chernausek S. D.
Published: December 2017

It has been proposed that insulin-like growth factor I (IGF-I) regulates fetal growth and differentiation. Plasma IGF-I concentrations correlate positively with fetal nutrient availability and newborn birth weights. To explore the hypothesis that hypoxemia decreases fetal growth by decreasing fetal IGF-I availability, we instrumented 14 fetal sheep with vascular catheters. At least 4 days after surgery, 10 fetuses were made acutely hypoxemic by infusing nitrogen into the maternal trachea for 3 h. Fetal blood oxyhemoglobin saturation decreased from 53 +/- 6 (SD) to 31 +/- 9%. Concomitantly, plasma IGF-I concentrations decreased from 91 +/- 11 to 67 +/- 10 ng/ml and IGF-I binding protein-1 concentration increased significantly, as assessed by ligand and Western blot analysis. Fetal IGF-I concentrations remained below control values throughout a subsequent recovery period (68 +/- 12 ng/ml at 6 h). In four control fetuses and in the ewes, plasma IGF-I concentrations were not significantly different from control values (97 +/- 18 and 181 +/- 18 ng/ml, respectively). These data support the hypothesis that decreases in fetal oxygen availability may decrease fetal growth by decreasing IGF-I production and availability.
Macrophage colony-stimulating factor increases hepatic macrophage content, liver growth, and lipid accumulation in neonatal rats
Authors: Pridans Clare , Sauter Kristin A. , Irvine Katharine M. , Davis Gemma M. , Lefevre Lucas , Raper Anna , Rojo Rocio , Nirmal Ajit J. , Beard Philippa , Cheeseman Michael , Hume David A.
Published: December 2017

Signaling via the colony-stimulating factor 1 receptor (CSF1R) controls the survival, differentiation, and proliferation of macrophages. Mutations in CSF1 or CSF1R in mice and rats have pleiotropic effects on postnatal somatic growth. We tested the possible application of pig CSF1-Fc fusion protein as a therapy for low birth weight (LBW) at term, using a model based on maternal dexamethasone treatment in rats. Neonatal CSF1-Fc treatment did not alter somatic growth and did not increase the blood monocyte count. Instead, there was a substantial increase in the size of liver in both control and LBW rats, and the treatment greatly exacerbated lipid droplet accumulation seen in the dexamethasone LBW model. These effects were reversed upon cessation of treatment. Transcriptional profiling of the livers supported histochemical evidence of a large increase in macrophages with a resident Kupffer cell phenotype and revealed increased expression of many genes implicated in lipid droplet formation. There was no further increase in hepatocyte proliferation over the already high rates in neonatal liver. In conclusion, treatment of neonatal rats with CSF1-Fc caused an increase in liver size and hepatic lipid accumulation, due to Kupffer cell expansion and/or activation rather than hepatocyte proliferation. Increased liver macrophage numbers and expression of endocytic receptors could mitigate defective clearance functions in neonates.

NEW & NOTEWORTHY This study is based on extensive studies in mice and pigs of the role of CSF1/CSF1R in macrophage development and postnatal growth. We extended the study to neonatal rats as a possible therapy for low birth weight. Unlike our previous studies in mice and pigs, there was no increase in hepatocyte proliferation and no increase in monocyte numbers. Instead, neonatal rats treated with CSF1 displayed reversible hepatic steatosis and Kupffer cell expansion.
Emerging potential benefits of modulating NAD+metabolism in cardiovascular disease
Authors: Matasic Daniel S. , Brenner Charles , London Barry
Published: December 2017

Nicotinamide adenine dinucleotide (NAD+) and related metabolites are central mediators of fuel oxidation and bioenergetics within cardiomyocytes. Additionally, NAD+is required for the activity of multifunctional enzymes, including sirtuins and poly(ADP-ribose) polymerases that regulate posttranslational modifications, DNA damage responses, and Ca2+signaling. Recent research has indicated that NAD+participates in a multitude of processes dysregulated in cardiovascular diseases. Therefore, supplementation of NAD+precursors, including nicotinamide riboside that boosts or repletes the NAD+metabolome, may be cardioprotective. This review examines the molecular physiology and preclinical data with respect to NAD+precursors in heart failure-related cardiac remodeling, ischemic-reperfusion injury, and arrhythmias. In addition, alternative NAD+-boosting strategies and potential systemic effects of NAD+supplementation with implications on cardiovascular health and disease are surveyed.
GABAergic contribution to the muscle mechanoreflex-mediated heart rate responses at the onset of exercise in humans
Authors: Teixeira André L. , Ramos Plinio S. , Samora Milena , Sabino-Carvalho Jeann L. , Ricardo Djalma R. , Colombari Eduardo , Vianna Lauro C.
Published: December 2017

Previous studies have indicated that central GABAergic mechanisms are involved in the heart rate (HR) responses at the onset of exercise. On the basis of previous research that showed similar increases in HR during passive and active cycling, we reasoned that the GABAergic mechanisms involved in the HR responses at the exercise onset are primarily mediated by muscle mechanoreceptor afferents. Therefore, in this study, we sought to determine whether central GABA mechanisms are involved in the muscle mechanoreflex-mediated HR responses at the onset of exercise in humans. Twenty-eight healthy subjects (14 men and 14 women) aged between 18 and 35 yr randomly performed three bouts of 5-s passive and active cycling under placebo and after oral administration of diazepam (10 mg), a benzodiazepine that produces an enhancement in GABAA activity. Beat-to-beat HR (electrocardiography) and arterial blood pressure (finger photopletysmography) were continuously measured. Electromyography of the vastus lateralis was obtained to confirm no electrical activity during passive trials. HR increased from rest under placebo and further increased after administration of diazepam in both passive (change: 12 ± 1 vs. 17 ± 1 beats/min, P < 0.01) and active (change: 14 ± 1 vs. 18 ± 1 beats/min, P < 0.01) cycling. Arterial blood pressure increased from rest similarly during all conditions ( P > 0.05). Importantly, no sex-related differences were found in any variables during experiments. These findings demonstrate, for the first time, that the GABAergic mechanisms significantly contribute to the muscle mechanoreflex-mediated HR responses at the onset of exercise in humans.

NEW & NOTEWORTHY We found that passive and voluntary cycling evokes similar increases in heart rate and that these responses were enhanced after diazepam administration, a benzodiazepine that enhances GABAA activity. These findings suggest that the GABAergic system may contribute to the muscle mechanoreflex-mediated vagal withdrawal at the onset of exercise in humans.
Neural control of nutritional and nonnutritional circuits in the dog hindpaw
Authors: Baker C. H. , Davis D. L. , Sutton E. T.
Published: December 2017

The neural control of blood flow and volume distribution between parallel nutritional and nonnutritional circuits has been investigated in the vascularly and neurally isolated dog hindpaw. Twelve paws were perfused by controlled pressure and ten paws were perfused by controlled flow via the cranial tibial artery. Venous outflow was measured and collected from the lateral saphenous vein. The superficial and deep fibular nerves and the tibial nerve were cut and individually stimulated, resulting in rates which at least doubled the blood flow resistance. Vascular volume changes were measured by injections of 51Cr-labeled red cells and 131I-labeled albumin. Tissue volume changes were measured by plethysmography. Capillary diffusion capacity was calculated from 86Rb extractions, and capillary filtration coefficients were determined. Superficial fibular nerve and deep fibular nerve stimulations apparently resulted in nonnutritional circuit constriction with resulting blood flow redistribution to nutritional circuits, possibly located in tissues other than the skin, e.g., adipose tissue. Tibial nerve stimulation caused no redistribution of blood flow between the two circuits presumably due to a uniform constriction of arteries and small vessel segments.
Mechanical responses of isolated dog cerebral arteries to reduction of external K, Na, and Cl
Authors: Toda N.
Published: December 2017

Reduction of the extracellular potassium concentration ([K+]0) caused a significant contraction in helical strips of dog cerebral arteries, but only a slight contraction, or none, in peripheral (coronary, mesenteric, renal, and femoral) arteries. The cerebroarterial contraction was abolished by treatment with ouabain or by substitution of Li+ for Na+, and suppressed by exposure to Ca2+-free media or verapamil. When one-half of the NaCl of the bathing solution was substituted with choline chloride a moderate, sustained contraction was produced in cerebral and peripheral arteries. Ouabain potentiated this contraction, whereas exposure to Ca2+-free media abolished it. Substitution of Na propionate, NaI, or Na acetylglycinate for one-half of the NaCl elicited a marked, transient contraction in only the cerebral arteries. This contractile response was potentiated by ouabain and markedly attenuated by exposure to Ca2+-free media. It may thus be concluded that cerebroarterial contractions induced by a reduction in [K+]0 are due to inhibition of the electrogenic Na+ pump, which results in depolarization of smooth muscle cells. Substitution of Cl- with less permeant anions appears to produce cerebroarterial contraction in association with depolarization of smooth muscle cells.
Progressive postnatal changes in sinus node response to atropine and propranolol
Authors: Woods W. T. , Urthaler F. , James T. N.
Published: December 2017

Postnatal development of autonomic control of heart rate was evaluated in the sinus nodes of isolated, perfused right atria obtained from 21 sibling puppies in three different litters between 3 and 11 wk postpartum. Age-related changes in response to autonomic blockade indicated that propranolol administered after atropine had its most profound direct depressive effect on less mature atrial cells. In the youngest sinus nodes the familial antimuscarinic action (increase in sinus rate) of atropine was regularly preceded by a brief period of cholinomimetic action (a marked slowing of sinus rate) in the youngest sinus nodes. These two opposing effects of atropine underwent a developmental change during the 2-mo study. The cholinomimetic action diminished, whereas the antimuscarinic action increased as a function of age.