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W D Rees

Publications and source records attributed to W D Rees.

At least 91 records · Page 5Linked to original sources

The effect of catecholamines and adenosine deaminase on the glucose transport system in rat adipocytes.

2-Deoxyglucose uptake (3 min) and 3-O-methylglucose transport (2 s) was measured in rat adipocytes preincubated with 5 microM epinephrine plus adenosine deaminase as described by Green (Green, A. (1983) FEBS Lett. 152, 261-264). 2-Deoxyglucose uptake was about 95% depressed in insulin-treated, but not in 'basal', cells preincubated with epinephrine plus adenosine deaminase for 60 min in broad agreement with Green's report. However, this depression was caused by a decrease in sugar phosphorylation rather than transport. In similarly incubated cells, transport of 3-O-methylglucose, a sugar analogue not phosphorylated in the adipocytes, was not affected by catecholamine plus adenosine deaminase. However, a decrease in transport of about 60% was observed both in the absence and the presence of insulin when the albumin concentration was high enough and the cell concentration low enough to prevent accumulation of free fatty acids in the medium. In addition, the insulin sensitivity with regard to hexose transport was markedly reduced. Transport was approximately doubled in cells incubated with 5 microM epinephrine in the absence of adenosine deaminase. Thus, epinephrine at a high concentration stimulates hexose transport in the absence of adenosine deaminase (presence of adenosine) whereas it inhibits both basal and insulin-stimulated transport in the presence of adenosine deaminase (absence of adenosine).

3-O-Methylglucose↗

A kinetic analysis of hexose transport in cultured human lymphocytes (IM-9).

3-O-Methyl-D-glucose transport across the plasma membrane of cultured human lymphocytes of the IM-9 line was followed for net entry into sugar-free cells (zero trans entry), net exit into sugar-free medium (zero trans exit) and for equilibration of labelled sugar in cells with the same sugar concentration in the intracellular water as in the medium (equilibrium exchange). The measurements were performed at 37 degrees C (pH 7.4). Equilibrium exchange of 1 mM 3-O-methylglucose (t 1/2 about 7 S) was exponential, suggesting a homogeneous cell suspension. Initial rates of transport showed a Michaelis-Menten dependency on the sugar concentration. The transport system was found to be asymmetric with the following kinetic parameters. Zero trans entry: Km = 2.8 mM, Vmax = 10.7 mM X min-1. Zero trans exit: Km = 9.5 mM, Vmax = 37.9 mM X min-1. Equilibrium exchange: Km = 9.9 mM, Vmax = 44.0 mM X min-1. Finally, the affinity constant for the internal site was measured as approx. 1.2 mM using the infinite cis protocol.

3-O-Methylglucose↗

Identification of a Na+,K+, Cl--cotransport protein of Mr 34 000 from kidney by photolabeling with [3H]bumethanide. The protein is associated with cytoskeleton components.

A polypeptide of Mr 34 000 is photolabeled with [3H]bumethanide after binding of this drug to membranes from the outer renal medulla and irradiation at 345 nm, a wavelength where bumethanide has an absorption maximum. Our data show that the polypeptide of Mr 34 000 is a component of the Na+/K+/Cl--cotransport system. The [3H]bumethanide binding protein is not extracted by concentrations of the nonionic detergent C12E8 that solubilizes 67% of the protein of the membranes including (Na+ + K+)-ATPase. This step increases the capacity for binding of [3H]bumethanide to 681 pmol/mg protein. Extraction of the binding protein requires high ionic strength suggesting that the Na+/K+/Cl--cotransport protein is associated with cytoskeleton components. This association may be important for control of the entry of NaCl into the cytoplasm and for cellular regulation of the rate of active transport of NaCl across the tubule cells in the thick ascending limb of Henles loop.

Animals↗

The fate of labelled glucose molecules in the rat adipocyte. Dependence on glucose concentration.

Isolated rat adipocytes were incubated with 15 nM [3-3H]glucose or 100 nM [U-14C]glucose with or without insulin and in the absence or presence of unlabelled glucose. Following a 2 h incubation with 15 nM [3-3H]glucose, about two thirds of the cell-associated 3H-labelled metabolic products were hydrophilic largely anionic intermediates and about one third was lipids. The equivalent values were 40 and 60%, respectively, when using 100 nM [U-14C]glucose. The only 14C-labelled metabolite escaping to the incubation medium was 14CO2, which accounted for about 15% of the rate of metabolism. Therefore, the rate of incorporation of 100 nM [U-14C]glucose into the cell-associated metabolites was quite a good measure of its net influx rate. The conversion of the two tracers to the sum of the metabolic products in cells treated with a maximally stimulating insulin concentration remained constant with glucose concentrations up to about 100 microM and then decreased progressively. The incorporation of radioactivity into the different metabolites varied markedly over the glucose concentration range 0-100 microM, presumably due to the saturation of different metabolic pools at different glucose concentrations. This variation was much less in cells not stimulated with insulin. Consequently, the maximal effect of insulin on the incorporation of the tracers into a given metabolite (e.g., labelled lipids) varied over the entire glucose concentration range. In addition, the apparent sensitivity (ED50) with respect to the incorporation into a given metabolite was also dependent on the glucose concentration.

Adipose Tissue↗

Cell loss from human, canine and guinea-pig gastric mucosa measured by DNA radioimmunoassay.

A DNA radioimmunoassay, sensitive in the range 25-1000 ng, has been developed to measure gastric mucosal cell loss. Validity of the assay was based on antibody specificity, absence of interference from gastric contents, parallel tracer displacement by dilutions of gastric and standard DNA, and crossover with colorimetric assay. With this assay, gastric DNA shedding was examined in two animal species and man. In the guinea-pig stomach, DNA loss was 20-100 micrograms/h and in the canine Pavlov pouch, 260-580 micrograms/h. In the canine Pavlov pouch stimulation of acid secretion by histamine and exposure to exogenous acid increased DNA output. In the normal human stomach DNA loss was 544 +/- 175 micrograms/h and in duodenal ulcer patients, 649 +/- 225 micrograms/h. DNA radioimmunoassay provides a sensitive and reproducible measure of cell exfoliation from the stomach and may be a useful tool for studying aspects of gastric mucosal defence.

Animals↗

Endoscopic diagnosis of a bleeding ileal carcinoid tumour.

Carcinoid tumour accounts for one per cent of all gastrointestinal neoplasms and has been reported in 0.5% of appendicectomy specimens. Local gastrointestinal complications occur infrequently and we report a case of repeated and massive gastrointestinal haemorrhage from a non-metabolically active carcinoid tumour of the distal ileum diagnosed by colonoscopy.

Carcinoid Tumor↗

Alkali secretion by isolated rabbit gastric mucosa: effects of non-steroidal anti-inflammatory drugs and prostaglandins.

The effects of aspirin, indomethacin and prostaglandins E2 and F2 alpha on the secretory and electrical activity of isolated rabbit gastric mucosa have been studied. Serosal side application of indomethacin (10(-5) M) or aspirin (3 X 10(-3) M) inhibited alkali secretion by fundic mucosa (mean +/- SE: 0.55 +/- 0.06 to 0.12 +/- 0.06 mumol X cm-2 X h-1, n = 6, p less than 0.01 and 0.28 +/- 0.06 to 0.11 +/- 0.03 mumol X cm-2 X h-1, n = 7, p less than 0.02 respectively) and antral mucosa (0.80 +/- 0.03 to 0.53 +/- 0.21 mumol X cm-2 X h-1, n = 5, p less than 0.01 and 0.75 +/- 0.23 to 0.47 +/- 0.20 mumol X cm-2 X h-1, n = 8, p less than 0.01 respectively). Mucosal and serosal side application of prostaglandin E2 or F2 alpha (10(-10) to 10(-4) M) had no effect on gastric alkali secretion. Serosal side 16,16 dimethyl E2 (10(-6) M) stimulated alkali secretion by fundic mucosa (0.90 +/- 0.20 to 1.50 +/- 0.30 mumol X cm-2 X h-1, n = 6, p less than 0.01) and abolished the inhibition of alkali production caused by indomethacin in fundic and antral mucosae. Inhibition of alkali secretion by aspirin was not modified by 16,16 dimethyl E2 pretreatment. The findings suggest that mucosal damage by aspirin and indomethacin may be mediated by inhibition of alkali secretion and that the protective action of various prostaglandins are only partly related to effects on this secretion.

Alkalies↗

The effects of an opiate agonist and antagonist on the human upper gastrointestinal tract.

Endogenous opiates may play a role in regulating intestinal function and we have investigated this possibility in the human upper gastrointestinal tract using loperamide as a locally-acting opiate agonist and naloxone as antagonist. Loperamide (4 mg) had no effect on the fasting pattern of motor activity in the antrum, duodenum and proximal jejunum. However, naloxone (40 micrograms kg-1 h-1, i.v.) abolished interdigestive motor activity in five of ten subjects and reduced antroduodenal motility index per cycle in all subjects (antrum: 1230 +/- 120 to 130 +/- 140 mm2; duodenum: 1120 +/- 150 to 225 +/- 160 mm2; P less than 0.005). These effects were not significantly affected by loperamide. Plasma motilin exhibited cyclic variation with significantly higher concentrations during antroduodenal phase 3 than during phases 1 and 2 of the interdigestive cycle. Plasma motilin was not altered by naloxone or loperamide, and neither drug had any effect on basal gastric acid secretion, duodenal output of amylase and bile salt or duodenogastric reflux. The results suggest that endogenous opiates regulate fasting motor activity in the human gastrointestinal tract but do not influence basal secretory function of the stomach, pancreas or biliary tree.

Adult↗

Effects of non-steroidal anti-inflammatory drugs and prostaglandins on alkali secretion by rabbit gastric fundus in vitro.

The effects of non-steroidal anti-inflammatory drugs and prostaglandins E(2) and F(2alpha) on the secretory and electrical activity of isolated rabbit fundic mucosa have been studied. Spontaneous acid secretion was inhibited by serosal side application of sodium thiocyanate (6x10(-2)M) and the resulting alkali secretion measured by pH stat tiration. Serosal side application of indomethacin (10(-5)M) or aspirin (3x10(-3)M) inhibited alkali secretion (0.55+/-0.06 to 0.12+/-0.06 mumol/cm(2)/h, n=6, p<0.01 and 0.28+/-0.06 to 0.11+/-0.03 mumol/cm(2)/h, n=7, p<0.02 respectively). Mucosal or serosal side prostaglandin E(2) (10(-5) to 10(-10)M) and F(2alpha) (10(-4) to 10(-10)M) failed to alter the rate of alkalinisation but secretion was significantly increased by serosal side 16,16-dimethyl-prostaglandin E(2) (10(-6)M) (0.90+/-0.20 to 1.50+/-0.30 mumol/cm(2)/h, n=6, p<0.01). Serosal side application of 10(-6)M prostaglandin E(2) to fundic mucosae pretreated with either aspirin (5x10(-3)M) or indomethacin (10(-5)M), to reduce endogenous E(2) formation, also failed to alter alkali secretion. Pretreatment of the mucosa with 16,16-dimethyl-E(2) (10(-6)M) abolished the inhibitory effect of indomethacin (10(-5)M) on alkali secretion (n=6) but did not modify the secretory response to aspirin (3x10(-3)M) (fall in alkali secretion with aspirin = 81+/-11% and with aspirin plus 16,16-dimethyl-E(2) = 72+/-10%, n=7). In the doses used, none of the prostaglandins or non-steroidal anti-inflammatory drugs altered transmucosal potential difference or electrical resistance. These results show that the damaging agents, aspirin and indomethacin, both inhibit gastric alkali secretion but that modes of action may differ. The observation that prostaglandins, E(2) and F(2alpha) failed to increase alkali production suggests that their protective activity against a variety of damaging agents as shown by others, may be mediated by another mechanism.

Alkalies↗

Human interdigestive and postprandial gastrointestinal motor and gastrointestinal hormone patterns.

Fasting gastrointestinal motor and hormone patterns were studied in 11 healthy volunteers. Cyclic motor activity was present in all subjects during fasting, but the duration and site of onset of each cycle were variable, even in the same subject. Fasting gastrin, GIP, and glucagon levels remained low and constant during the 8-hr study, while plasma motilin levels exhibited cyclic variation in 7 of the 11 subjects. Achlorhydria (induced with cimetidine in 5 of the 11 subjects) did not alter the pattern of fasting motor activity or plasma motilin. In the remaining six subjects, the effect of liquid nutrient meals was examined. Ingestion of a sodium chloride bolus failed to disrupt fasting cyclic activity, while all nutrient-containing solutions inhibited gastric phase-2 motor activity, the duration of inhibition being longest for the mixed and lipid meals. All nutrient meals released GIP, while only protein and mixed meals released gastrin, and the lipid meal released motilin. Our study confirms the rhythmicity of interdigestive motor cycles in man and demonstrates their lack of dependence on gastric acid secretion and some relationship to motilin cycles in certain individuals as determined by radioimmunoassay. Transition from fasting to fed pattern (after liquid meals) is characterized by the inhibition of phasic gastric pressure changes in the antrum and the development of irregular activity in the intestine, similar in pattern to fasting phase 2. Because the duration of interruption of the gastric interdigestive pattern by meals depends on their nutrient content, we conclude that dietary composition may be a major determinant of the fasting-fed motor balance in man.

Achlorhydria↗

A demonstration of bicarbonate production by the normal human stomach in vivo.

Bicarbonate secretion by the stomach may play a role in gastric mucosal protection, and we have therefore examined bicarbonate secretion in the human stomach. Gastric bicarbonate production and contamination with salivary and duodenal bicarbonate was measured in healthy volunteers using an intubation technique. The stomach and duodenum were perfused with nonabsorbable markers and the pH, PCO2, marker, and amylase concentrations measured in 10-min gastric and duodenal aspirates. These measurements enabled calculation of gastric bicarbonate content and the amount of bicarbonate contributed by saliva and duodenogastric reflux. Acid secretion was suppressed by intravenous cimetidine. Validation studies demonstrated a good correlation between instilled and calculated recovered bicarbonate (r = 0.97, P less than 0.001, N = 6), and marker recovery was consistent in each subject. Over a 6-h period, gastric pH ranged from 6 to 7 and PCO2 from 20 to 40 mm Hg. Gastric bicarbonate output stabilized at 326--392 mumol/hr and mean bicarbonate concentration ranged from 2.3 to 20.0 mmol/liter. Approximately two thirds of this bicarbonate was free, 11% was derived from duodenogastric reflux, and 3% from swallowed saliva. This study demonstrates secretion of bicarbonate by the human stomach in vivo at a rate equivalent to 10--20% of basal acid secretion.

Adult↗

Studies of acid and alkaline secretion by rabbit gastric fundus in vitro: effect of low concentrations of sodium taurocholate.

The effect of low concentrations of sodium taurocholate on the secretory and electrical activity of isolated rabbit fundic mucosa has been studied. Fundic mucosa maintained a stable potential difference (10.2 +/- 0.6 mV, n = 70) and electrical resistance (85 +/- 6 omega . cm-2, n = 70) and the majority of preparations spontaneously secreted acid (2.85 +/- 0.31 mumol . cm-2 . h-1, n = 70). Histamine (10(-5) and 10(-4) M) and carbachol (10-4 M) increased acid secretion, and these responses were prevented by cimetidine (10(-3) M) and atropine (10(-5) M), respectively. Mucosal application of taurocholate (10(-4) and 10(-3) M) increased net acidity without altering electrical activity. This response exhibited tachyphylaxis, was not altered by pretreating the tissues with cimetidine (10(-3) M), atropine (10(-5) M), or somatostatin (10(-6) M), and occurred in mucosas maximally stimulated by histamine. Sodium thiocyanate (6 x 10(-2) M, serosal side) inhibited spontaneous acid secretion revealing net alkalinization (0.83 +/- 0.05 mumol . cm-2 . h-1, n = 58) that was completely inhibited by anoxia and potassium cyanide (10(-2) M) and markedly reduced by 2,4-dinitrophenol (10(-4) M). Some fundic preparations spontaneously secreted alkali (1.20 +/- 0.20 mumol . cm-2 . h-1, n = 6) after an initial period of acid secretion. Mucosal-side taurocholate (10(-3) M) converted net alkali secretion by both thiocyanate-treated and spontaneously-secreting mucosas to acid secretion without affecting electrical conductance. These secretory responses may be implicated in the pathogenesis of gastric mucosal damage by bile salts.

Animals↗

Interdigestive motor activity in patients with systemic sclerosis.

Fasting antral, duodenal, and jejunal motor activity and plasma motilin and pancreatic polypeptide were studied in 14 patients with systemic sclerosis, 6 and 9 without clinical evidence of small bowel involvement, and 8 healthy control subjects. Normal interdigestive motor activity was present in control subjects and patients without intestinal involvement. However, cyclic motor activity was absent in 3 of the patients with intestinal disease and the motility index per interdigestive cycle (or per 6-h recording period in those without cyclic activity) was significantly less in the antrum (181 +/- 103 mm2 compared with 760 +/- 86 and 1116 +/- 96 mm2 for patients without involvement and healthy control subjects, respectively), duodenum (153 +/- 101 mm2 compared with 1425 +2- 186 nd 1055 +/- 241 mm2), and jejunum (268 +/- 131 mm2 compared with 1166 +/- 97 and 1105 +/- 128 mm2). Metoclopramide and bethanechol significantly increased motor activity at the three sites in all subjects but the magnitude of the metoclopramide response was less in patients with intestinal involvement. Fasting concentrations of motilin and pancreatic polypeptide exhibited cyclic variation with peak values occurring during phase 3 of the interdigestive cycle. Plasma motilin during each phase of motor activity was significantly higher in patients with scleroderma, with or without intestinal involvement, than in control subjects. The abnormal motor activity demonstrated here indicates a possible mechanism by which intestinal stasis and bacterial overgrowth could occur and by which clinical disturbances of intestinal transit might arise.

Adult↗