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R A Coleman

Publications and source records attributed to R A Coleman.

At least 73 records · Page 4Linked to original sources
PubMed1995

Characterization of the PGE receptor subtype mediating inhibition of superoxide production in human neutrophils.

1. The aims of this study were to characterize the EP receptor subtype mediating the inhibition of superoxide anion generation by formyl methionyl leucine phenylalanine (FMLP)-stimulated human neutrophils, and to test the hypothesis that adenosine 3':5'-cyclic monophosphate (cyclic AMP) is the second messenger mediating the inhibition of the neutrophil by prostaglandin (PG)E2. 2. PGE2 (0.001-10 microM) inhibited FMLP (100 nM)-induced O2-generation from human peripheral blood neutrophils in a concentration-dependent manner, with an EC50 of 0.15 +/- 0.03 microM, and a maximum effect ranging from 36-84% (mean inhibition of 68.7 +/- 2.5%, n = 32). 3. The EP2-receptor agonists, misoprostol, 11-deoxy PGE1, AH13205 and butaprost, all at 10 microM, inhibited O2- generation, causing 95.5 +/- 2.9%, 56.8 +/- 5.2%, 37.1 +/- 6.6% and 18.9 +/- 4.4% inhibition respectively, the latter two being much less effective than PGE2. Similarly, the EP1-receptor agonist, 17-phenyl PGE2 (10 microM), and the EP3/EP1-receptor agonist, sulprostone (10 microM), also inhibited O2- generation, causing 32.2 +/- 7.0% and 15.3 +/- 3.4% inhibition respectively. 4. The non-selective phosphodiesterase inhibitor, isobutyl methylxanthine (IBMX, 0.25 mM) inhibited the FMLP response by 54.5 +/- 5.0%. In addition, IBMX shifted concentration-effect curves for PGE2, misoprostol, 11-deoxy PGE1, butaprost, and AH 13205 to the left, to give EC50s of 0.04 +/- 0.03 (n = 13), 0.07 +/- 0.03 (n = 4), 0.08 +/- 0.03 (n = 4), 0.33 +/- 0.13 (n = 4) and 0.41 +/- 0.2 microM (n = 3) respectively, allowing equieffective concentration-ratios (EECs, PGE2 = 1) of 11.5, 5.3, 50.7 and 12.7 to be calculated. This agrees well with the relative potencies of these agonists at EP2 receptors.5. By contrast, even in the presence of IBMX (0.25 mM), sulprostone and 17-phenyl PGE2 were only effective at the highest concentration (10 microM), and gave EECs of > 700 and 486 respectively, suggesting that EP1 or EP3 receptors are not involved.6. The selective type IV phosphodiesterase inhibitor, rolipram at 2 and 10 nM did not inhibit the FMLP response, but at the higher concentration of 50 nM, it decreased the FMLP response by 46.6 +/-7.3%.However, rolipram shifted concentration-effect curves for PGE2 to the left to give EC50s of 0.06 +/-0.022,0.015 +/- 0.0, 0.012 +/- 0.006 microM at 2, 10 and 50 nM respectively, compared to the control EC50 of0.27+/- 0.09 microM for PGE2.7. The EP4/TP receptor blocking drug, AH 23848B (10 microM, 10 min) did not inhibit 02- generation by PGE2, but was found to potentiate significantly the effect of PGE2 at the lower concentrations of PGE2 tested (0.001-0.1 microM).8. The adenylate cyclase inhibitor, SQ 22,536 (0.1 mM, 2 min) reduced PGE2-induced inhibition of 02-production, giving an EC50 in the absence of SQ 22,536 of 0.24 +/- 0.1, and 1.9 +/- 1.1 AM in its presence.9. These results suggest that inhibition of superoxide generation by PGE2 is mediated by stimulation ofEP2 receptors and activation of adenylate cyclase, leading to the elevation of intracellular levels of cyclic AMP.

Adenine↗
PubMed1995

Characterization of the prostanoid receptors mediating constriction and relaxation of human isolated uterine artery.

1. This study was undertaken to characterize pharmacologically the prostanoid receptor subtypes mediating constriction and relaxation of human isolated uterine artery. 2. U-46619 was a potent constrictor agonist on human uterine artery (EC50 [95% CL] = 3.5 [1.8-6.7] nM). Prostaglandin E2 (PGE2), PGF2 alpha, PGD2 and PGI2 only weakly constricted the uterine artery, being at least 100 times less potent than U-46619. The PGE2 and PGI2 constrictor effects may be modified by the potent dilator effects of these compounds. A number of agonists which show selectivity for FP-, DP- and EP-receptors including ICI 81008, BW 245C, sulprostone, rioprostil and butaprost, failed to cause any constriction at concentrations up to 30 microM. 3. Constrictor responses induced by all agonists tested were reduced or abolished by the TP-receptor blocking drugs, GR 32191 and EP 092. pA2 estimates for both antagonists versus U-46619 were 8.50, values which are consistent with their affinities at TP-receptors. 4. In preparations pre-constricted with phenylephrine (1 microM) both PGI2 and PGE2 were potent relaxant agonists. The selective IP-receptor agonists, cicaprost and iloprost, also dilated human uterine artery and were approximately 10 fold more potent than PGI2. The EP2-receptor agonists, butaprost and rioprostil and the selective DP-agonist, BW 245C, were at least 100 fold weaker than PGI2 and PGE2 suggesting that neither DP- nor EP2 receptors were involved. 5. We conclude that TP-receptors mediate constriction, whereas IP- and possibly EP4-receptors mediate relaxation of human uterine artery.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
PubMed1995

Linoleic and linolenic acids are selectively secreted in triacylglycerol by hepatocytes from neonatal rats.

To determine whether specific fatty acids are metabolized differently by neonatal liver, hepatocyte cultures from neonatal (age: 5, 11, and 21 days) and adult rats were incubated with radiolabeled 18:1, 18:2, or 18:3. At each age, the rate of oxidation was highest for 18:3 and lowest for 18:1. Conversely, esterification was highest for 18:1 and lowest for 18:3. Fatty acid esterification was of the order: day 5 > day 11 > adult > day 21. When incubations contained each of two of the above fatty acids, one radiolabeled and the other not, 18:1 inhibited oxidation of radiolabeled 18:2 by up to 45% in neonatal hepatocytes. In addition, added 18:1 increased glycerolipid accumulation from 18:2 and 18:3. Under these conditions, the relative proportion of triacylglycerol secreted in the medium, compared with that accumulated in the cells, was two- to fourfold higher for day 11 and 21 rat hepatocytes. The results suggest that a specific mechanism exists in the livers of neonatal rats to spare n-3 and n-6 fatty acids from oxidation and instead secrete them in triacylglycerol.

Aging↗
PubMed1995

Developmental regulation of the catalytic subunit of the apolipoprotein B mRNA editing enzyme (APOBEC-1) in human small intestine.

Apolipoprotein (apo) B mRNA editing is a site-specific cytidine deamination reaction responsible for the production of apoB-48 in mammalian small intestine. This process is mediated by an enzyme complex that includes the catalytic subunit, APOBEC-1. In the present study, it is shown that the developmental regulation of apoB mRNA editing in fetal human small intestine is closely mirrored by accumulation of APOBEC-1 mRNA. Similar results were obtained using Caco-2 cells, the data further suggesting that culture of these cells under conditions previously shown to promote differentiation produce an earlier and more marked induction of APOBEC-1 mRNA abundance. Complementary analysis of APOBEC-1 protein accumulation using immunocytochemical localization reveals its appearance to be temporally coordinated with the accumulation of APOBEC-1 mRNA and its distribution to be confined to villus-associated enterocytes. Previous studies demonstrated a close temporal association between the development of triglyceride synthesis and apoB mRNA editing in the rat liver and small intestine. Analysis of fatty acid CoA ligase, monoacylglycerol acyltransferase, and diacylglycerol acyltransferase activity in preparations of human liver and small intestine demonstrates activity of all three enzymes in the late first and early second trimester, suggesting that certain aspects of complex lipid biosynthesis in the human fetal small intestine and liver are regulated developmentally. The cues that modulate the post-transcriptional regulation of fetal human small intestinal apoB gene expression may thus include both temporal programming and events related to the emergence of lipid transport capability.

APOBEC-1 Deaminase↗
PubMed1995

Metabolism of fenbufen by cultured 3T3-L1 adipocytes: synthesis and metabolism of xenobiotic glycerolipids.

The storage of xenobiotic compounds as glycerolipids and their subsequent mobilization was studied using fenbufen and differentiated 3T3-L1 cells in culture. Fenbufen was taken up from the incubation medium and incorporated into triacylglycerol, diacylglycerol, and phospholipids. The triacylglycerol was susceptible to digestion by pancreatic lipase. The xenobiotic phospholipid contained three species, one of which behaved as fenbufenoyl phosphatidylcholine as judged by TLC, HPLC, choline analysis, and mass spectroscopy. After incubation with radioactive fenbufen for 18 h, the cells were transferred to a chase medium where radioactivity was lost from the cells and appeared in the medium. The rate was three times higher when 10 microM isoproterenol was present; insulin had no effect. Non-esterified fenbufen and analogues of mono- and di-acylglycerol were secreted. Monofenbufenoylglycerol was characterized by its ability to be used as a substrate by purified monoacylglycerol acyltransferase. When oleic acid was used in place of fenbufen, the majority of the radioactivity released in a chase experiment was the non-esterified acid (over 90%) and neither mono- nor di-acylglycerol was detected. These data indicate that 3T3-L1 adipocytes can synthesize fenbufen-containing lipids and release them into the medium on hormonal stimulation. The secretion of mono- and di-acylglycerols may have unforeseen pharmacological or toxicological implications.

3T3 Cells↗
PubMed1994

Hepatic monoacylglycerol acyltransferase is regulated by sn-1,2-diacylglycerol and by specific lipids in Triton X-100/phospholipid-mixed micelles.

The lipid cofactor requirement of hepatic monoacylglycerol acyltransferase (MGAT) (EC 2.3.1.22) was studied in Triton X-100/lipid-mixed micelles. Anionic phospholipids and anionic lysophospholipids stimulated MGAT activity, whereas fatty acids and sphingosine inhibited enzyme activity. Phosphatidic acid was a potent activator, stimulating MGAT 11-fold at 4.2 mol %. Kinetic studies revealed that phosphatidic acid, with an apparent Ka of 0.26 mol %, was a better activator than phosphatidylserine, phosphatidylinositol, or cardiolipin. Of the anionic lysophospholipids, lysophosphatidic acid was a better activator than lysophosphatidylserine, stimulating maximally at less than 3 mol %. Oleate was a more potent inhibitor (Ki, 2.4 mol %) than sphingosine (Ki, 18.3 mol %). The dependence of MGAT on sn-2-monoacylglycerol was not cooperative in the absence or presence of anionic phospholipids, oleic acid, or sphingosine. The apparent Km for sn-2-monoC18:1-glycerol was 1.24 mol % in the presence of maximally activating phospholipid and 0.19 mol % when phospholipid was omitted. MGAT's product sn-1,2-diacylglycerol was a weaker activator than the anionic phospholipids, but the effects of diacylglycerol and phospholipid were additive. Activation by sn-1,2-diC18:1-glycerol was highly cooperative with a Hill coefficient of 3.6. Activation was specific for the sn-1,2-stereoisomer; neither 1,3-diacylglycerol nor the ether analogs of sn-1,2- or 1,3-diacylglycerol were activators. Since several of the lipid modulators of MGAT activity are intracellular second messengers, these data suggest the possibility that regulatory links exist between signal transduction and the synthesis of complex lipids via the monoacylglycerol pathway.

Acyltransferases↗
PubMed1994

Glycogen storage disease in adults.

OBJECTIVE: To identify complications amenable to prevention in adults with glycogen storage disease (GSD) types Ia, Ib, and III and to determine the effect of the disease on social factors. DESIGN: Case series and clinical review. SETTING: Referral medical centers in the United States and Canada. PATIENTS: All patients with GSD-Ia (37 patients), GSD-Ib (5 patients), and GSD-III (9 patients) who were 18 years of age or older. MEASUREMENTS: Ultrasound or radiographic studies identified liver adenomas, nephrocalcinosis, or kidney stones. Radiographic studies identified osteopenia. Reports of the clinical examination, serum chemistry results, and social data were obtained. RESULTS: For patients with GSD-Ia, problems included short stature (90%), hepatomegaly (100%), hepatic adenomas (75%), anemia (81%), proteinuria or microalbuminuria (67%), kidney calcifications (65%), osteopenia or fractures or both (27%), increased alkaline phosphatase (61%) and gamma-glutamyltransferase (93%) activities, and increased serum cholesterol (76%) and triglyceride (100%) levels. Hyperuricemia was frequent (89%). Patients with GSD-Ib had severe recurrent bacterial infections and gingivitis. In patients with GSD-III, 67% (6 of 9) had increased creatinine kinase activity. Four of these patients had myopathy and cardiomyopathy. CONCLUSIONS: For GSD-Ia, hyperuricemia and pyelonephritis should be treated to prevent nephrocalcinosis and additional renal damage. For GSD-Ib, granulocyte-colony-stimulating factor may prevent bacterial infections. For GSD-III, more data are required to determine whether the myopathy and cardiomyopathy can be prevented. Most of the patients with GSD-I and GSD-III had 12 or more years of education and were either currently in school or employed.

Adult↗
PubMed1994

Formoterol on airway smooth muscle and human lung mast cells: a comparison with salbutamol and salmeterol.

Formoterol, like salbutamol and salmeterol, relaxed isolated preparations of guinea-pig trachea and human bronchus, and inhibited antigen-induced mediator release from human lung fragments in a concentration-related fashion. In each case, these actions were mediated through beta 2-adrenoceptors, with formoterol being 50-120-fold more potent than salbutamol, and 2-27-fold more potent than salmeterol. The duration of action of formoterol was longer than that of salbutamol in all preparations, but was markedly shorter than that of salmeterol, whose actions persisted for many hours despite continuous or extensive washing of the tissues. In conscious guinea-pigs, inhaled formoterol, salbutamol and salmeterol all caused dose-related inhibition of histamine-induced bronchoconstriction. Formoterol was again more potent (10-20-fold) than either salbutamol or salmeterol. However, while the actions of a threshold-effective dose of formoterol persisted for less than 3 h, somewhat longer than those of salbutamol (< 1.5 h), an equivalent dose of salmeterol was active for at least 6 h. Therefore, while formoterol is a potent beta 2-adrenoceptor agonist in vitro and in vivo, and is consistently longer-acting than salbutamol, its duration of action is markedly shorter than that of salmeterol.

Albuterol↗
PubMed1994

Erythromycin, lincosamides, peptidyl-tRNA dissociation, and ribosome editing.

Inaccurate protein synthesis produces unstable beta-galactosidase, whose activity is rapidly lost at high temperature. Erythromycin, lincomycin, clindamycin, and celesticetin were shown to counteract the error-inducing effects of streptomycin on beta-galactosidase synthesized in the antibiotic-hypersensitive Escherichia coli strain DB-11 Met-. Newly synthesized beta-galactosidase was more easily inactivated by high temperatures when synthesized by bacteria partially starved for arginine, threonine, or methionine. Simultaneous treatment with erythromycin or lincomycin yielded beta-galactosidase that was inactivated by high temperatures less easily than during starvation alone, an effect attributed to stimulation of ribosome editing. When synthesized in the presence of canavanine, beta-galactosidase was inactivated by high temperature more easily but this effect could not be reversed by erythromycin. The first arginine in beta-galactosidase occurs at residue 13, so the effect of erythromycin during arginine starvation is probably to stimulate dissociation of erroneous peptidyl-tRNAs of at least that length. Correction of errors induced by methionine starvation is probably due to stimulation of dissociation of erroneous peptidyl-tRNAs bearing peptides at least 92 residues in length. All the effects of erythromycin or the tested lincosamides on protein synthesis are probably the result of stimulating the dissociation from ribosomes of peptidyl-tRNAs that are erroneous or short.

Anti-Bacterial Agents↗
PubMed1994

Adipose monoacylglycerol:acyl-coenzyme A acyltransferase activity in the white-throated sparrow (Zonotrichia albicollis): characterization and function in a migratory bird.

Although migrating birds use stored triacylglycerol as their primary fuel for flight, they must retain sufficient stores of omega 6 and omega 3 fatty acids to sustain reproduction after the spring migration. Hepatic monoacylglycerol:acyl-coenzyme A acyltransferase (EC 2.3.1.22) (MGAT) activity is associated with physiological periods in which lipolysis and beta-oxidation are prominent, and it may also play a role in the selective retention of certain essential fatty acids. Therefore, we characterized MGAT activity in adipose tissue from the white-throated sparrow (Zonotrichia albicollis), a migratory bird. MGAT specific activity from adipose tissue and liver, respectively, was 22.2 +/- 7.27 and 0.79 +/- 0.35 nmol/min/mg of total particulate protein. Activity did not vary seasonally or between male and female birds. Specific activity increased 4.3-fold in the presence of 75 micrograms of phosphatidylcholine and phosphatidylserine (1:1, w/w). MGAT acylated sn-1(3)-monooleoylglycerol, sn-2-monooleylglycerol ether and sn-1(3)-monooleylglycerol ether at 7.5, 5.7 and 1.7%, respectively, of the rate observed with sn-2-monooleoylglycerol. An initial lag phase observed at low concentrations of palmitoyl-CoA was corrected by adding 2 mM MgCl2, Mg(NO3)2 or CaCl2, suggesting a requirement for divalent cations. MGAT acylated sn-2-monolinolenoylglycerol and sn-2-monolinoleoylglycerol in preference to sn-2-monooleoylglycerol. Specificity of MGAT for sn-2-monoacylglycerols and the probable enhanced affinity fo sn-2-monoacylglycerols of specific acyl chains may allow selected omega 6 and omega 3 fatty acids to be retained within the adipocyte, while nonessential fatty acids are released for beta-oxidation in flight muscles.

Acyltransferases↗
PubMed1994

A novel inhibitory prostanoid receptor in piglet saphenous vein.

A range of prostanoid agonists were tested for activity on isolated ring preparations of piglet saphenous vein. The selective TxA2-mimetic (TP-receptor agonist), U-46619, contracted the preparation in a concentration-related fashion. These contractions were inhibited by the TP-receptor blocking drug, GR32191B, producing a pA2 of 7.8 (slope = 1.6). Prostanoid-induced relaxant responses were studied on preparations which had been pre-contracted using an EC60 concentration of phenylephrine (mean EC60 = 0.97 microM), in the presence of GR32191B (1 microM), to block contractile TP-receptors. Under these conditions, PGD2, PGE2, PGF2 alpha, PGI2, and U-46619, all caused concentration-related relaxation. PGE2 was the most potent agonist (EC50 = 0.23nM), whereas, all of the other agonists were at least 1,000-fold weaker, providing strong evidence for the presence of inhibitory EP-receptors. The selective synthetic EP-agonists, sulprostone (EP1/EP3) and AH13205X (EP2), were next tested for relaxant activity. While both compounds caused concentration-related relaxant activity, they were respectively 6,000 and 11,000-fold less potent than PGE2. The potent TP-receptor blocking drugs, AH22921X and AH23848B, were both weak antagonists of PGE2 but not isoproterenol-induced relaxant responses of piglet saphenous vein in a concentration-related fashion. These two compounds had pA2 values against PGE2 of 5.3 and 5.4 respectively, with regression slopes not significantly different from unity. In contrast, neither compound at a concentration of 30 microM had any antagonist activity against prostanoid-induced effects on guinea-pig fundus (EP1), rabbit ear artery (EP2) or guinea-pig vas deferens (EP3). In conclusion, the piglet saphenous vein contains TP-receptors mediating smooth muscle contraction, and a PGE2-specific (EP) receptor mediating relaxation. The inhibitory EP-receptor does not appear to be of the EP1, EP2 or EP3-subtypes, and appears therefore to be a novel subtype which we tentatively term EP4, and the potent TP-receptor blocking drugs, AH22921X and AH23848B, appear to be weak, but specific EP4-receptor blocking drugs.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
PubMed1994

The effects of PACAP and VIP on guinea pig tracheal smooth muscle in vitro.

Neither pituitary adenylate cyclase activating polypeptide-38 (PACAP) nor its homologue, vasoactive intestinal polypeptide (VIP), contracted guinea pig isolated trachea (GPT), but on preparations contracted with KCl (40 mM), both caused concentration-related relaxation (3 nM-3 microM, VIP IC50 = 72 nM, PACAP IC50 = 224 nM). Relaxant curves to PACAP were slower in reaching a maximum than those to VIP (approximately 150 and 50 min, respectively). The protease inhibitors, phosphoramidon (1 microM), leupeptin (50 microM), bestatin (100 microM), soya bean trypsin inhibitor (1 microM), and aprotinin (5 microM), together caused a small enhancement of relaxations to VIP, but not to PACAP. The VIP antagonist, [4-Cl-D-Phe6, Leu17]VIP (1-10 microM), did not inhibit the relaxation to either peptide, but did cause large contractions, which were enhanced by protease inhibition. These findings demonstrate that PACAP relaxes GPT in a similar manner to VIP.

Animals↗
PubMed1994

The effects of IAPP and CGRP on guinea pig tracheal smooth muscle in vitro.

Neither the novel peptide, islet amyloid polypeptide (IAPP), nor its homologue, calcitonin gene-related peptide (CGRP), contracted guinea pig isolated trachea (GPT), but on preparations contracted with KCl (40 mM), both caused concentration-related relaxation (1 nM-3 microM). Relaxant curves to both were shallow and slow in development, with clear maxima not being obtained. The protease inhibitors, phosphoramidon (1 microM), leupeptin (50 microM), bestatin (100 microM), soya bean trypsin inhibitor (1 microM), and aprotinin (5 microM), together had no effect on relaxations to CGRP, but depressed those to IAPP. Aprotinin appeared to be responsible for this depression. The specific CGRP antagonist, CGRP(8-37), 1-10 microM, had no effect on relaxations to either peptide. These findings demonstrate that IAPP relaxes GPT in a similar manner to CGRP.

Amyloid↗
PubMed1994

ADH-induced recycling of fluid-phase marker from endosomes to the mucosal surface in toad bladder.

In the toad urinary bladder, the reversal of antidiuretic hormone (ADH) stimulation results in the endocytosis of apical membrane water channels, along with any fluid-phase marker present in the mucosal bathing solution. We have loaded vesicles with horseradish peroxidase (HRP), then restimulated the bladders and measured the reappearance of endocytosed HRP in the mucosal bath. HRP-loaded bladders that were restimulated showed HRP release that peaked sharply within 15 min after restimulation. Varying the interval between loading and restimulation did not vary HRP release significantly. Restimulation with forskolin gave HRP release values similar to ADH. The amount of HRP released correlated with the magnitude of water permeability induced. The demonstration that fluid-phase markers can be recycled from endosomes to the apical surface in a hormone-dependent fashion indicates that endocytosed membrane, containing water channels, is able to rapidly recycle back to the surface in response to hormone restimulation. In addition, marker release declined progressively with repeated restimulation, totaling < 30% of the retrieved amount. This result indicates that a relatively large proportion of the retrieved marker reaches a nonrecycling compartment.

Animals↗
PubMed1994

Long-term regulation of collecting duct water permeability: freeze-fracture analysis of isolated perfused tubules.

Terminal inner medullary collecting duct (IMCD) segments from water-restricted rats have high osmotic permeabilities despite the absence of vasopressin (AVP). We performed freeze-fracture analysis of individual IMCD segments from such animals following measurement of their water permeability (Pf). IMCD segments from control rats did not have a high Pf in the absence of AVP (88 +/- 15 microns/s) and had a low incidence of E-face intramembrane particle (IMP) clusters (9.6 +/- 2.7 clusters/100 microns 2). Segments exposed to 0.1 nM AVP in vitro had enhanced Pf (1,060 +/- 210 microns/s) and cluster incidence (122 +/- 33 clusters/100 microns 2). IMCD segments isolated from rats dehydrated for 48 h and perfused without AVP exposure had an elevated Pf (605 +/- 71 microns/s) and a high incidence of clusters (166 +/- 36 clusters/100 microns 2). There also was an increase in the number of single particles between clusters in tubules from dehydrated rats (2.5-fold) and in AVP-treated tubules (3.6-fold). These findings indicate that IMP clusters are associated with high water permeability in tubules from dehydrated animals independent of continued AVP exposure. The increased incidence of particles between clusters suggests that water channels may also occur outside of cluster domains.

Animals↗