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Acid secretion by isolated canine gastric mucosa.

1. An isolated gastric mucosal preparation from the dog stomach which is capable of acid secretion is described. Average values for normal resting potential difference (p.d.) was 49 +/- 2 mV (mucosal side negative with respect to the serosal side), short-circuit current (Isc) was 172 +/- 4 muA and resistance (R) was 285 +/- 6 omega.cm2. Low rates of spontaneous acid secretion (0-0.58 muequiv/cm2.hr) were present initially but following short-circuiting of the tissue these values decreased to low levels (less than 0.1 muequiv/cm2.hr) within an hour. 2. Histamine in doses exceeding 10(-6)M stimulated acid secretion, increased Isc and decreased R. Concentrations ranging from 10(-5) to 8 X 10(-4)M produced maximal secretion. The maximal secretory rate achieved was 4.24 +/- 0.35 muequiv/cm2.hr. 3. Pentagastrin (10(-8)M) and acetylcholine (10(-6)--10(-5)M) also stimulated acid secretion with a lower maximal secretion as compared to histamine stimulation. These concentrations of pentagastrin and acetylcholine did not alter histamine stimulated acid secretion. Higher concentrations of pentagastrin (10(-6)M) and acetylcholine (10(-4) M) reversibly inhibited acid secretion of histamine stimulated mucosa. 4. These results demonstrate that there are many similarities between in vitro and in vivo findings on the dog stomach, indicating the great potential of the in vitro dog gastric mucosa for studies on the mechanism of action and interaction of gastric secretagogues.

Acetylcholine↗

Effect of taurohyodeoxycholic acid, a hydrophilic bile salt, on bile salt and biliary lipid secretion in the rat.

Taurohyodeoxycholic acid is a natural 6 alpha-hydroxylated bile acid with an apparent hydrophilicity intermediate between those of tauroursodeoxycholic and taurocholic acids. We investigated in the rat the hepatobiliary metabolism, choleretic properties, and biliary maximum secretory rate (SRmax) of taurohyodeoxycholic in comparison with these two bile salts. Each compound was infused intravenously, at a rate increased in a stepwise manner from 100 to 300 nmol/min/100 g body wt, in bile salt-depleted bile fistula rats. The three bile salts appeared rapidly starting with the infusion and increased to represent more than 95% of the total bile salts. No apparent biliary metabolites were formed. All the bile salts caused a dose-dependent increase in bile flow and biliary lipid output. The absolute increase in bile flow was lower in rats infused with taurohyodeoxycholic acid, yet the volume of bile formed per nanomole of secreted bile salt was 13.8 nl for taurohyodeoxycholic, 6.4 nl for tauroursodeoxycholic acid, and 10.9 nl for taurocholic. The SRmax values were 1080, 3240, and 960 nmol/min/100 g, respectively. At all infusion rates, taurohyodeoxycholic acid caused a greater (P < 0.001) secretion of biliary lecithin compared to the other bile salts. There were no significant differences in the biliary secretion of cholesterol and proteins. Electron microscopy showed the recruitment of vesicles and lamellar bodies around and within bile canaliculi. In conclusion, taurohyodeoxycholic promotes a biliary lecithin secretion greater than expected from physicochemical predictions, representing a novel secretory property with potential pharmacological relevance.

Animals↗

Participation of Ca2+ and calmodulin in rat pancreatic enzyme secretion induced by secretin, forskolin, and dibutyryl cyclic AMP.

The role of Ca2+ and calmodulin in stimulation of the rat pancreatic acini induced by secretin, forskolin, and dibutyryl cyclic AMP (dbcAMP) was studied using W-7, a calmodulin antagonist, and a low Ca2+ medium. The time course of amylase secretion was studied in a perfusion system using dispersed rat pancreatic acini. The amylase release patterns of each secretagogue were as follows: a biphasic amylase release pattern under the stimulation of secretin, a one peak pattern during the stimulation of forskolin and a rapid response after cessation of the stimulation, and a gradual increased pattern during the stimulation of dbcAMP followed by a rapid response. The amylase release under the stimulation by forskolin and dbcAMP was slightly weaker as compared with that of secretin stimulation. The amylase secretion stimulated by secretin (5 X 10(-7) M), forskolin (50 microM), and dbcAMP (2 mM) was inhibited by W-7 (50 microM). In a low Ca2+ medium (4.7-5.1 X 10(-6) M), the secretory rate did not increase during the stimulation by secretin, forskolin, and dbcAMP, and a rapid amylase response remained after cessation of the stimulation of forskolin and dbcAMP. The pretreatment with EDTA (1 mM) suppressed both the gradual amylase release and the rapid response induced by dbcAMP in a low Ca2+ medium. These results suggested that each secretagogue, via cyclic AMP (cAMP), induced a different amylase secretory pattern dependent on an intracellular Ca2+ content, and was mediated by the Ca2+-calmodulin complex.

1-Methyl-3-isobutylxanthine↗

Electrolyte secretion by the isolated cat pancreas during replacement of extracellular bicarbonate by organic anions and chloride by inorganic anions.

1. The effect of replacing extracellular bicarbonate and chloride by other anions on the volume and composition of secretin-stimulated pancreatic juice has been analysed in the isolated, perfused cat pancreas. 2. The anions of some aliphatic carboxylic acids were able partially to substitute for bicarbonate in sustaining pancreatic secretion. The order of effectiveness was: acetate greater than proprionate greater than butyrate greater than formate. 3. The rate of secretion in the presence of 25 mM-acetate was 42% of that achieved with 25 mM-bicarbonate. The concentration of acetate in the secretion varied with flow rate, reaching a maximum of 120 mM at high flow rates and declining at lower flow rates, with reciprocal changes in chloride concentration. Bicarbonate was always present in the secretion at a concentration of 5--7 mM. 4. Inorganic anions were able totally or partially to substitute for chloride in sustaining secretion. In relation to chloride, their degree of effectiveness was: chloride = bromide = or greater than nitrate greater than iodide greater than sulphate greater than methyl sulphate greater than isethionate. Those anions which had no effect on secretion rate (i.e. bromide and nitrate) also had no effect on the bicarbonate concentration of the secretion and themselves appeared in the secretion in place of chloride. Those anions which inhibited secretion increased the bicarbonate concentration in the secretion in proportion to the degree of inhibition they caused (i.e. the increase was greatest with isethionate). 5. When perfusate chloride was only partially replaced by bromide or iodide the ratios of chloride: bromide and chloride: iodide in the secretion were approximately equal to those in the perfusate. 6. The carbonic anhydrase inhibitor acetazolamide reduced secretory rate and bicarbonate concentration when added to normal perfusion fluid or chloride-substituted fluids, but had no effect following replacement of perfusate bicarbonate by acetate. 7. These observations illustrate that an extracellular source of permeant anions is required for optimal pancreatic bicarbonate secretion to occur. This may indicate the participation of an anion exchange carrier in the transport events responsible for this secretory process.

Acetazolamide↗

A proposed mechanism for increased tear-film osmolarity in contact lens wearers.

Contact lenses decrease corneal sensitivity and increase tear-film osmolarity. To determine whether the decrease in corneal sensitivity is responsible for the increase in tear-film osmolarity, we studied tear-film osmolarity in normal rabbits and rabbits with surgically induced keratoconjunctivitis sicca before and after the instillation of 0.5% proparacaine. Instillation of proparacaine resulted in a significant increase in tear-film osmolarity. Decreased corneal sensitivity, with a resultant decrease in tear secretory rates, is the most likely cause for increased tear-film osmolarity in wearers of hard contact and extended-wear soft contact lenses.

Animals↗

An in vivo preparation for measurement of plasma protein and lysozyme output in the ferret tracheal lumen.

An in-vivo ferret tracheal preparation has been developed to study the appearance in the liquid-filled trachea of fluorescein-labelled plasma proteins (FLP) and of lysozyme from submucosal gland serous cells. In order to investigate the influence of nervous activity on the appearance of FLP and lysozyme in the tracheal lumen, the effects of intraluminal bradykinin (an inflammatory mediator and sensory nerve stimulant), intraluminal capsaicin (a stimulant of C-fibres) and electrical stimulation of the cut peripheral end of the right cervical vagus nerve have been measured. Vagal stimulation (10 V, 10 Hz, 1 ms, 90-120 s) increased the secretory rate of lysozyme. It had no effect on FLP rate of output. Intraluminal bradykinin (100 microM) produced a small but significant increase in FLP output but had no effect on lysozyme secretion. Intraluminal capsaicin (33 microM) had no effect on FLP output and had variable effects on lysozyme output. Tracheal pressure was increased by vagal stimulation but was unaffected by bradykinin and capsaicin. Thus, bradykinin increases plasma protein output, probably by an action on the epithelium, whilst vagal stimulation and capsaicin stimulate submucosal glands. This method could be used to determine the factors which alter the rate of movement of plasma proteins into the airway lumen and the secretion of submucosal glands in vivo.

Animals↗

The importance of conjugation in biliary secretion of ursodeoxycholate and 7-ketolithocholate in the rat.

The biliary maximum secretory rate (SRmax.) of glycoursodeoxycholate in the bile fistula anaesthetized rat was about five times that of unconjugated ursodeoxycholate. Likewise, the biliary SRmax. of tauro-7-ketolithocholate was more than three times that of unconjugated 7-ketolithocholate. The SRmax. of 7-ketolithocholate infused with taurine (to avoid depletion of the taurine pool) was still significantly lower than that of exogenous tauro-7-ketolithocholate. These data support the hypothesis that, for bile acids that are poorly water soluble, the maximal biliary secretion rate is dependent on conjugating capacity, which is the limiting step in the overall transport from plasma into bile.

Animals↗

Comparison of gastric mucosal blood flow as determined by aminopyrine clearance and gamma-labeled microspheres.

Gastric mucosal blood flow was simultaneously determined by aminopyrine clearance and gamma-labeled microspheres (15 +/- 5 mu in diameter) in anesthetized dogs prepared with a chambered segment of stomach greater curvature. Paired flow measurements were made in 11 dogs (n = 28) secreting in response to intravenous histamine (1mug per kg per min), in 11 (n = 21) nonsecreting dogs given intravenous isoproterenol (0.5 or 1.0 mug per kg per min), and in 9 (n = 10) dogs given no drugs to stimulate secretion or blood flow (resting dogs). Eight additional injections were done in dogs receiving various combinations of isoproterenol and histamine. Isotonic HCl was maintained on the mucosal surface during all experiments. Regression analysis demonstrated a highly significant linear correlation between clearance and microsphere-measured flow in the histamine (P less than 0.001, r = 0.96) and isoproterenol (P less than 0.001, r = 0.78) experiments, with clearance averaging 83% of microsphere flow during histamine stimulation but only 25% during isoproterenol. The relationship between clearance and microsphere flow was not significantly different for the resting and isoproterenol experiments. Mucosal perfusion measured by microspheres was about 5 times the resting value for both histamine and isoproterenol-stimulated dogs. Perfusion calculated from aminopyrine clearance averaged 46, 38, and 90% of the microsphere value in the resting, isoproterenol, and histamine experiments, respectively. Pooled data from secreting dogs demonstrated a fairly constant ratio of microsphere-measured flow to clearance (1.25 +/- 0.06 mean +/- SEM), regardless of the secretory rate. Our results indicate that aminopyrine clearance reflects only a small fraction of mucosal blood flow in the nonsecreting stomach, even in the presence of exogenous acid.

Aminopyrine↗

Hepatic triglyceride secretion in relation to lipogenesis and free fatty acid mobilization in fasted and glucose-refed rats.

Plasma triglyceride concentrations were significantly lowered by a single feeding of glucose to rats that had been fasted for 22 hr. Three feedings of glucose produced a similar effect. In the glucose-refed animals mobilization of free fatty acids from adipose tissue was impaired more rapidly than hepatic lipogenesis was restored from its low fasting level. These effects of glucose were shown by both a 50% fall in plasma free fatty acid concentration and an 84% decrease in free fatty acid release by isolated epididymal fat pads within 30 min after a single refeeding of glucose. Hepatic lipogenesis from either acetate-1-(14)C or glucose-U-(14)C was not restored even after glucose had been fed three times at hourly intervals. Triton-induced hypertriglyceridemia was used to measure the hepatic triglyceride secretory rate; it was found that glucose refeeding decreased this rate in all but one of several experiments. This decreased secretion rate was sufficient to account for the nearly complete disappearance of triglyceride in very low density lipoproteins (d < 1.019) that occurred within 1 hr after a single glucose intubation.

Acetates↗

Determination of aldosterone secretion rate utilizing mixed mode and high performance liquid chromatography.

A simpler method for determining aldosterone secretion rate (ASR) has several applications. High performance liquid chromatography (HPLC) has several advantages over traditional chromatographic methods for purification to constant specific activity of aldosterone liberated from its 18-glucuronide by acid hydrolysis. We found it necessary to introduce several modifications to remove urochromes before HPLC. Two methods for determining ASR were developed. With Method A a more traditional initial procedure was followed, and Sephadex LH-20 chromatography allowed removal of considerable urochromes before HPLC. However, aldosterone recovery was improved with Method B, which employed several bonded phase silica derivatives (Sepralytes) and a PBE 94 column to remove urochromes before HPLC. With this procedure the Sephadex LH-20 chromatography was not required. Aldosterone purification to constant specific activity was achieved by HPLC on a diol column with a normal phase system, and quantification was performed by RIA. ASR determinations were equivalent with both methods. This methodology should be applicable to other steroid secretory rate determinations and to applications involving purification of steroid conjugates.

Aldosterone↗

A technique for the in vitro perfusion of bovine parathyroid glands.

A system for the in vitro perfusion of bovine parathyroid glands was developed. The glands were perfused at 1 ml/minute with oxygenated Hanks balanced salt solution buffered with sodium bicarbonate to a pH of 7.4 at 37 degrees C. These glands remained functionally stable for seven to eight hours. The secretory rate of immunoreactive parathyroid hormone varied inversely with the calcium concentration of 3.0 to 6.0 mg/100 and became nonsuppressible at calcium concentrations above 6.5 mg/100 ml. Therefore, the glands are thought to be responsive to stimuli. A loss of functional stability can usually be detected by an increase in perfusion pressure. The samples of effluent containing immunoreactive parathyroid hormone remained stable (immunoreactively) for six months when stored at -70 degrees C. This system provides a valid method for monitoring dynamic, minute-to-minute changes in parathyroid secretory activity.

Animals↗

Inhibition of acid secretion from vagally innervated and denervated gastric pouches by (Gln4)-neurotensin.

In dogs with innervated fundic pouches an intravenous infusion of (Gln4)-neurotensin, 60 pmol X kg-1 X min-1, reduced the pentagastrin-stimulated acid secretion to 40% of the control secretory rate. After vagal denervation of the fundic pouches the same dose of (Gln4)-neurotensin did not significantly change the secretion. These findings suggest that (Gln4)-neurotensin does not act directly on parietal cells but inhibits acid secretion at a presynaptic level.

Animals↗

Is salt reabsorption in the human sweat duct subject to control?

There is a time lag between the beginning of sweat secretion and fully effective production of fluid of minimum salt content. It is suggested that changes in permeability to water or electrolytes have to be effected and may account for the time lag. The composition of thermal and pilocarpine sweat indicates that pharmacological stimulation does not wholely reproduce the physiological mechanism: pilocarpine sweat contains more sodium and more cyclic AMP, irrespective of the secretory rate. The sweat obtained from patients with cystic fibrosis has been compared with that from normal children: the concentrations of cyclic AMP and cyclic GMP are the same. Defective sodium reabsorption in the patients is therefore unlikely to be due to inadequate or excessive synthesis of these cyclic nucleotides.

Absorption↗

Long-term organ culture of large numbers of human fetal pancreata: analysis of their insulin secretion.

Cultured human fetal pancreas has been transplanted into diabetic man in an attempt to cure the metabolic disorder. However, the capacity of large numbers of these organs to secrete insulin in organ culture has not been reported previously. This report sets out the characteristics of 321 human fetal pancreases of gestational age 12-20 weeks obtained over a period of 34 months, 295 of these being maintained in organ culture. Average insulin secretion was constant at 2.22 +/- 0.35 mU/plate/day over a 90-day period, the maximal duration of culture possible. Explants were lost because of infection or reduced insulin secretion. Practical guidelines for obtaining and maintaining viable explants in culture were established as follows: Insulin secretion had to be greater than 0.1 mU/plate/day. Secretory rates of this order were associated with a positive insulinogenic response to theophylline. Prostaglandin induction, suction curettage, and hysterotomy were equally suitable as methods of termination of pregnancy. The pancreas had to be obtained within 4 hours of termination of pregnancy. The tissue had to be diced into explants within a further 4 hours.

Fetus↗

Purinergic P2y receptors stimulate renin secretion by rat renal cortical slices.

These experiments were designed to characterize P2 purinergic receptor-mediated effects on renin secretion, using the rat renal cortical slice preparation. 2-Methylthio ATP (10-500 microM) and ATP (100-500 microM) stimulated renin secretion in a concentration-dependent manner and 2-methylthio ATP was the more potent. By contrast, alpha, beta-methylene ATP (0.1-500 microM) had no effect on renin secretion. This order of potency (2-methylthio ATP > ATP > alpha, beta-methylene ATP) indicates that activation of the P2y subclass of purinergic receptors stimulates renin secretion. Theophylline did not antagonize the effect of 2-methylthio ATP, which suggests that the effect was not due to a conversion of 2-methylthio ATP to 2-methylthio adenosine, followed by activation of P1 purinergic receptors. In contrast, N omega-nitro-I-arginine methyl ester both antagonized the basal renin secretory rate and blocked the stimulating effects on renin secretion of 2-methylthio ATP. Because N omega-nitro-l-arginine methyl ester antagonizes the production of nitric oxide by endothelial cells, these results suggest that nitric acid stimulates basal renin secretion in this experimental preparation and that increased production of it mediates the stimulating effects on renin secretion of activation of P2y purinergic receptors.

Adenosine Triphosphate↗

Prostaglandins and renin release: II. Assessment of renin secretion following infusion of PGI2,E2 and D2 into the renal artery of anesthetized dogs.

The influence of intra-renal infusions of prostaglandin (PG) I2, PGE2 and PGD2 on renin secretion and renal blood flow was investigated in renally denervated, beta-adrenergic blocked, indomethacin treated dogs with unilateral nephrectomy. All three prostaglandins when infused at doses of 10(-8) g/kg/min and 10(-7) g/kg/min resulted in marked renal vasodilation. Renin secretory rates increased significantly with both PGI2 and PGE2 at the 10(-8) g/kg/min and 10(-7) g/kg/min infusion rates in a dose dependent manner. However, PGD2 was inactive. At 10(-7) g/kg/min, PGI2 infusions resulted in systemic hypotension indicated recirculation of this prostaglandin. These findings suggest that PGI2 should be included among the cyclooxygenase derived metabolites of arachidonic acid to be considered as possible mediators of renin release.

Animals↗

Growth and growth hormone secretion after treatment for childhood non-Hodgkin's lymphoma.

The aim of this study was to evaluate the growth and growth hormone (GH) secretion, as assessed by the rate and pattern of secretion, in patients in remission from non-Hodgkin's lymphoma (NHL) who had been treated with corticosteroids and intense chemotherapy. None of the patients had received cranial irradiation. Twelve children were investigated yearly by taking 24-hour GH profiles starting 1 year from the time of diagnosis. The mean age at onset of the disease was 7.5 years. Another 12 young adults were studied in a cross-sectional manner 4.1-21.3 years (mean, 9.0 years) after diagnosis of NHL. The mean age at onset of the disease was 10.7 years. The median height velocity was significantly decreased during the 1st year following diagnosis (standard deviation scores [SDS] -0.15, P < .001), especially during the first 3 months (SDS -0.75, P < .001) when the most intense treatment was given. During the 2nd year height velocity was still somewhat reduced (SDS -0.13, P < .001). However, there was no reduction in final attained height. Spontaneous GH secretion, in terms of both secretory rate and pulsatile pattern, was evaluated by measuring integrated GH concentrations in 20-minute blood samples collected over a 24-hour period. The plasma GH concentrations were transformed into GH secretion rates by means of a deconvolution technique. Fourier time series analysis was applied to determine possible disturbances of rhythmicity of the GH secretion. The GH secretion rate and the pulsatile pattern of secretion in the NHL patients were similar to those of the reference population of pubertal matched healthy controls. There was no influence of the age at diagnosis or of the time from diagnosis of NHL on the GH secretion rate. Growth impairment in children with a malignant disease treated only with steroids and chemotherapy is therefore probably not caused by disturbed GH secretion, but rather by direct interference with bone growth of the cytotoxic drugs used. There was no significant influence on weight gain during the treatment period so an indirect effect of chemotherapy on bone growth through interference with adequate nutrition seems unlikely. However, GH secretion was not evaluated during the period of growth retardation, and therefore a transient deficiency was not excluded.

Adolescent↗

Effects of increased O2 and CO2 on acid secretion by dogfish gastric mucosa in vitro.

The gastric mucosa of the dogfish (Squalus acanthias), as usually prepared for in vitro chambered experiments, shows a secretory rate (JH) of about 2 mueq/cm2-h, but a potential difference (PD) of zero. Raising PCO2 from 0.05 to 0.1 atm increases JH by 40% and causes the development of a PD of about 2 mV, mucosal surface positive. Increasing PO2 from 0.9 to 1.9 atm in a hyperbaric chamber (at constant PCO2 = 0.1 atm) doubles JH and increases PD to 5 mV. Transepithelial resistance falls by 20% at high PO2. It appears that the dogfish gastric mucosa, like that of the frog, is rate limited by CO2 diffusion into the tissue from the usual 5% mixture and is also rate limited by the usual O2 levels (unlike the frog), presumably due to its thicker structure and higher O2 consumption. The mucosal-positive PD, which is reversed from all other mucosae studied, is readily explained by separate electrogenic H+ and Cl- pumps, but less readily by schemes embodying a neutral HCl pump. It is not yet known whether the hyperbaric conditions are sufficient to ensure O2 sufficiency.

Animals↗