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M L Cohen

Publications and source records attributed to M L Cohen.

At least 163 records · Page 9Linked to original sources
PubMed1991

Protein kinase C translocation in rat stomach fundus: effects of serotonin, carbamylcholine and phorbol dibutyrate.

In an effort to characterize serotonergic receptor activation in rat stomach fundus, the potential role of protein kinases, more specifically protein kinase C (PKC), in serotonin-induced contraction of rat stomach fundus was examined. Staurosporine, a potent, but nonselective, inhibitor of protein kinases, attenuated basal, membrane-bound PKC activity in rat stomach fundus (IC50 = 10 nM). Although staurosporine (3-100 nM) produced a concentration-dependent inhibition of contractions elicited by serotonin (which does not increase phosphatidylinositol hydrolysis in the fundus), carbamylcholine (an agent stimulating phosphatidylinositol hydrolysis), and phorbol 12,13-dibutyrate (PDBu; a phosphatidylinositol-independent activator of PKC translocation), it was a more potent inhibitor of contractions produced by serotonin and PDBu than by carbamylcholine. Potassium chloride-induced contractions were attenuated minimally by staurosporine. These results raised the possibility that serotonin might exert an effect on protein kinase activity by a phosphatidylinositol-independent mechanism. Focusing on PKC, serotonin's ability to translocate PKC from cytosol to membrane in rat fundus was examined. Concentrations of serotonin (0.1-10 microM) which maximally contracted rat fundus did not translocate PKC. However, PDBu (10 nM-1 microM) and carbamylcholine (0.1-10 microM) significantly increased membrane-bound PKC activity. These results: 1) demonstrate that translocation of PKC occurred in rat stomach fundus in response to some, but not all, contractile agonists; 2) are consistent with the possibility that contraction of rat stomach fundus by carbamylcholine and PDBu may be related to increased membrane-bound PKC activity; and 3) indicate that serotonin-induced contraction, although potently blocked by staurosporine, did not result from PKC translocation in the rat stomach fundus.

Alkaloids↗
PubMed1990

Congenital tricuspid valve stenosis with atrial septal defect and left anterior fascicular block.

Congenital tricuspid valve stenosis in the absence of other valvular abnormalities is rare. In this report we describe a patient with congenital tricuspid valve stenosis, ostium secundum atrial septal defect, and electrocardiographic left anterior fascicular block, who presented with paradoxical emboli. This case, as well as previous case reports, suggests that congenital tricuspid stenosis with ostium secundum atrial septal defect is associated with left anterior fascicular block.

Adult↗
PubMed1990

The ambivalent effects of early and late administration of mannitol in cold-induced brain oedema.

This study was undertaken in order to determine whether early administration of mannitol is different from late administration in its effect on brain oedema. Cold-induced brain oedema, which was confirmed by high resolution CT scan, was produced in 2 groups of cats. In group one mannitol was given early (90 minutes after injury); in group two 3-4 hours after the injury (late). Repeated CT scans following mannitol administration showed that the early group exhibited significantly greater dehydration (p less than 0.0001) while the late group showed significant hydration, in the lesioned hemisphere. The contralateral control hemisphere responded to mannitol with similar dehydration effect in both groups.

Animals↗
PubMed1990

Inhibition of serotonin-amplified human platelet aggregation by ketanserin, ritanserin, and the ergoline 5HT2 receptor antagonists-LY53857, sergolexole, and LY237733.

Human platelets are known to possess 5HT2 receptors which, when activated, amplify the aggregation response produced by other aggregating agents. Several 5HT2 receptor antagonists, including ketanserin and ritanserin, are known to antagonize serotonin-mediated aggregation of human platelets. In the present study, we document the ability of three ergoline 5HT2 receptor antagonists, LY53857, sergolexole, and LY237733, to antagonize the serotonergic component of the human platelet aggregation response. Potencies of the ergoline esters (LY53857 and sergolexole) and the ergoline amide (LY237733) to inhibit serotonin-amplified platelet aggregation responses were similar to the potencies of ketanserin and ritanserin under the conditions of our study. Furthermore, all five 5HT2 receptor antagonists were capable of fully inhibiting the serotonergic component of the platelet aggregation response. In contrast to these potent ergoline esters and amides, 1-isopropyl dihydrolysergic acid (up to 10(-5)M), a putative metabolite of the ergoline esters, was ineffective under these in vitro conditions. These data are consistent with the high potency of these ergolines as antagonists of 5HT2 receptors and further support the involvement of 5HT2 receptors on human platelets in the amplifying response to serotonin.

Adult↗
PubMed1990

Bradykinin-induced contractions of canine prostate and bladder: effect of angiotensin-converting enzyme inhibition.

Alpha-adrenergic, cholinergic, and serotonergic receptor-mediated contractile responses have been well characterized in the genitourinary tissues of several mammalian species. The present study characterizes the in vitro contractile responsiveness of canine bladder and prostate to the peptides, bradykinin, angiotensin I, and angiotensin II. All preparations contracted to 0.15 M KCl. Bradykinin elicited contractile responses in both prostate (10(-10) to 10(-7) M) and bladder (10(-10) to 10(-6) M). In both tissues, angiotensin II produced minimal responses and angiotensin I failed to elicit contractions. The potent angiotensin converting enzyme (ACE) inhibitor, enalaprilic acid [MK-422] (10(-6) M) increased the contractile response to the prostate to bradykinin two-fold while having no effect on bradykinin-induced contractions in the bladder. Enalaprilic acid did not affect the contractile responses of the two tissues to angiotensin I or angiotensin II. The canine urogenital tissue contractile responses to bradykinin, angiotensin I, and angiotensin II may have relevance to human physiology. Previous studies have demonstrated that human prostatic tissue, specifically benign prostatic hyperplasia (BPH), has the highest concentration of ACE activity of tissues evaluated. Bradykinin is a potent peptidergic contractile agent in canine bladder and prostate. The activity of enalaprilic acid to amplify the bradykinin-induced contractions in the canine prostate is consistent with high levels of ACE in the tissue. These data confirm the sensitivity of the canine prostate to bradykinin and report for the first time, the ability of bradykinin to induce contractions in the prostate. These studies support the possibility that bradykinin may be involved in mediating micturition under normal and pathological states such as infravesical obstruction secondary to BPH. Furthermore, the results from these investigations in canine urogenital tissues, if applicable to humans, suggest that urinary function be closely monitored in patients receiving ACE inhibitor therapy.

Angiotensin I↗
PubMed1990

Canine, but not rat bladder contracts to serotonin via activation of 5HT2 receptors.

Serotonin is a biogenic amine that can exert multiple effects on smooth muscle, including smooth muscle of the genitourinary tract; effects that may be species dependent. The present study using isolated tissues documents potent contractile responses to serotonin in canine bladder smooth muscle. Contractile responses to serotonin in canine bladder could be mimicked by alpha-methyl serotonin, a selective 5HT2 receptor agonist. In fact, although alpha-methyl serotonin was slightly less potent as a contractile agonist relative to serotonin, the contractile response to alpha-methyl serotonin was more persistent as evidenced by a greater recovery time to resting force following washout. In contrast, the 5HT1A receptor agonist, 8-OH-DPAT and the 5HT3 selective agonist, 2-methyl serotonin, did not markedly contract canine bladder. These data establish that contractile responses to serotonin in the canine bladder are mediated by activation of 5HT2 receptors. We further demonstrated that the 5HT2 receptor antagonist, LY53857, potently inhibited the contractile response to both serotonin and alpha-methyl serotonin in the canine bladder consistent with agonist activation of 5HT2 receptors. In contrast to the potent response to serotonin observed in the canine bladder, rat bladder preparations did not markedly contract in response to serotonin, alpha-methyl serotonin, 8-OH-DPAT, or 2-methyl serotonin. Thus, these studies reinforce the marked species variability in responsiveness to serotonin and indicate that contraction to serotonin in the canine bladder is mediated by activation of the 5HT2 receptor.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1990

(8 beta)-6-methylergoline amide derivatives as serotonin antagonists: N1-substituent effects on vascular 5HT2 receptor activity.

A series of (8 beta)-6-methylergoline amide derivatives was synthesized with various alkyl substituents in the N1-position in order to evaluate their effectiveness in blocking vascular 5HT2 receptors. The influence of both the N1 substituent and amide derivative proved to be of great importance on binding affinities to vascular 5HT2 receptors. Within each series of amides, however, maximum affinity was achieved with an N1-isopropyl substituent (14, 18, 26, 38, and 41; all with 2.7-50 times greater affinity than their N1-H analogues), with the exception of two cases (22 and 37) in the cyclohexylamide derivatives wherein N1-methyl equalled the isopropyl in potency. Other than these exceptions, affinities followed the pattern of H less than Me less than Et less than iPr, with potencies falling off with larger alkyl substituents.

Amides↗
PubMed1990

Synthesis and biochemical evaluation of tritium-labeled 1-methyl-N-(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)-1H-indazole-3-carboxa mide, a useful radioligand for 5HT3 receptors.

The advent of potent, highly selective 5HT3 receptor antagonists has stimulated considerable interest in 5HT3 receptor mediated physiology and pharmacology. To permit detailed biochemical studies regarding interaction of the indazole class of serotonin (5HT) antagonists with 5HT3 receptors in multiple tissues, we synthesized 1-methyl-N-(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)-1H-indazole- 3-carboxamide (LY278584, compound 9) in high specific activity, tritium-labeled form. This radioligand was selected as a synthetic target because of its potency as a 5HT3-receptor antagonist, its selectivity for this receptor viz a viz other 5HT-receptor subtypes, and the ability to readily incorporate three tritia via the indazole N-CH3 substituent. Alkylation of N-(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)-1H-indazole-3-carboxamide (8) with sodium hydride and tritium-labeled iodomethane, followed by HPLC purification, resulted in [3H]-9 with a radiochemical purity of 99% and a specific activity of 80.5 Ci/mmol. This radioligand bound with high affinity to a single class of saturable recognition sites in membranes isolated from cerebral cortex of rat brain. The Kd was 0.69 nM and the Bmax was 16.9 fmol/mg of protein. The specific binding was excellent, and accounted for 83-93% of total binding at concentrations of 2 nM or less. The potencies of known 5HT3-receptor antagonists as inhibitors of [3H]-9 binding correlated well with their pharmacological receptor affinities as antagonists of 5HT-induced decreases in heart rate and contraction of guinea pig ileum, suggesting the central recognition site for this radioligand may be extremely similar to or identical with peripheral 5HT3 receptors.

Animals↗
PubMed1990

Influence of the 60-megadalton plasmid on adherence of Escherichia coli O157:H7 and genetic derivatives.

The adherence of enterohemorrhagic Escherichia coli serotype O157:H7 and various genetic derivatives to Henle 407 intestinal and HEp-2 epithelial cell lines was examined by light and electron microscopy. The parent outbreak strain, 7785, harbors a 60-megadalton serotype-specific plasmid designated pO157 and adhered to both cell lines, as determined by light microscopy. A plasmidless derivative, 2-45, showed reduced adherence to both cell lines. After being labeled with Tn801, pO157 was transformed into E. coli C600, E. coli HB101, and E. coli GH42, and back into 2-45. Both E. coli C600 and HB101 transformants adhered weakly; full adherence was restored to the 2-45(pO157::Tn801) transformant. Transmission electron microscopy (EM) demonstrated the intimate attachment of HB101(pO157::Tn801) to Henle 407 cells which formed cuplike structures and areas of possible actin polymerization adjacent to adhering bacterial cells; scanning EM further extended these observations. EM studies of E. coli O157:H7 strains were hampered by extensive intestinal cell damage, presumably due to the action of Shiga-like toxins. EM also demonstrated that 7785 and its plasmidless derivative 2-45 were piliated and that no pili were apparent on HB101(pO157::Tn801) or GH42//(pO157::Tn801). The plasmid pO157 appears to modify the eucaryotic cell adherence of E. coli O157:H7 and to confer that adherence on E. coli HB101 through surface structures other than pili. These findings, when compared with other published reports, also suggest similarities between enterohemorrhagic and enteropathogenic E. coli adherence properties.

Bacterial Adhesion↗
PubMed1990

Immunologic characterization of a 35-kilodalton recombinant antigen of Mycobacterium tuberculosis.

A 35-kilodalton (kDa) recombinant antigen (35-kDa antigen) produced by Escherichia coli JM107 carrying DNA from Mycobacterium tuberculosis was purified and immunologically examined by in vivo and in vitro methods. A monoclonal antibody (2B2) was produced against the 35-kDa antigen. The protein was purified from the insoluble fraction of the recombinant E. coli strain by either affinity chromatography with the 2B2 monoclonal antibody or preparative isoelectric focusing. In enzyme-linked immunosorbent assay and Western blot (immunoblot) analyses, antibody to 2B2 reacted with whole-cell sonic extracts of M. tuberculosis and other slowly growing mycobacteria but not with two rapid growers, M. chelonae and M. fortuitum. An injection series totaling less than 1 mg of purified protein without adjuvant elicited a humoral response in guinea pigs. In one guinea pig, 10 micrograms of purified protein injected intradermally elicited both a humoral and a cell-mediated response. Results of these studies suggest that the 35-kDa antigen is a membrane-associated protein that stimulates both humoral and cell-mediated immune responses and should be evaluated as a vaccine candidate.

Animals↗
PubMed1990

Alterations of intercellular junctions induced by hypoxia in canine myocardium.

To delineate potential structural mechanisms of impaired cell coupling induced by hypoxia in canine myocardium, we characterized derangements in intercellular junctions and alterations in the space constant in strips of ventricular epimyocardium before and after selected intervals of hypoxia in vitro. Tissue samples were analyzed morphometrically with transmission and freeze-fracture electron microscopy. Space constants in control tissues averaged 1.61 +/- 0.47 mm (mean +/- SD). After 30 and 60 min of hypoxia, space constants declined by 0.24 +/- 0.22 and 0.32 +/- 0.17 mm, respectively (P less than 0.05 vs. control in each case). Impaired coupling was not reversible with reoxygenation. Focal pathological separation of intercalated disk membranes was observed after 30 min of hypoxia, but morphometric analysis demonstrated no reduction in gap junction surface density to account for uncoupling after 30 min of hypoxia. However, after 60 min of hypoxia, gap junction surface density was reduced by 45%. Quantitative analysis of freeze-fractured gap junction replicas after 30 min of hypoxia revealed a significant decrease in P-face particle diameter from 8.51 +/- 1.64 nm in control tissues to 7.25 +/- 1.33 nm (P less than 0.01) with no further change at 60 min. Thus impaired coupling at 30 min is likely related to a change in the gap junction particle. Further uncoupling after 60 min of hypoxia is likely related, in addition, to reduced gap junction surface density. These results suggest that alterations in P-face particles and gap junction surface density are important determinants of progressive cellular uncoupling induced by hypoxia.

Animals↗
PubMed1990

Experimental cardiac tamponade: correlation of pressure, flow velocity, and echocardiographic changes.

Seven episodes of experimental cardiac tamponade were induced in five anesthetized closed-chest dogs. Simultaneous pericardial and intracavitary pressures were synchronized with superior vena caval and transvalvular pulsed-Doppler flow tracings. The earliest indication of tamponade was the development of a negative transmural right atrial pressure that occurred during early ventricular diastole and was associated with echocardiographic evidence of right atrial collapse. This was also associated with reversal of diastolic flow in the superior vena cava and with diminished early diastolic flow velocity across the tricuspid as well as the mitral valve. During more advanced cardiac tamponade, the transmural right atrial pressure became negative during both early and late ventricular diastole as well as during isovolumic ventricular systole. This was associated with a disappearance of early diastolic ventricular filling and right ventricular diastolic collapse as observed on two-dimensional echocardiography. In hypotensive cardiac tamponade (cardiac output diminished by 70%), the decreased transmural right atrial pressure that developed during ventricular systole was accompanied by diminished antegrade flow in the superior vena cava. In advanced and hypotensive tamponade, ventricular filling occurred mainly during atrial contraction.

Animals↗
PubMed1990

5-Hydroxytryptamine3 receptors mediate tachycardia in conscious instrumented dogs.

Intravenously administered 5-HT and the 5-HT3 selective agonist, 2CH3-5-HT, and the 5-HT2 selective agonist, alpha-CH3-5-HT, transiently increased heart rate in conscious, instrumented dogs. 5-HT, alpha-CH3-5-HT and 2CH3-5-HT increased systolic blood pressure in conscious dogs. The increase in blood pressure produced by alpha-CH3-5-HT was blocked by the 5-HT2 selective antagonist, LY53857, supporting a role for vascular 5-HT2 receptors in the pressor response to these amines. In contrast, LY53857 did not antagonize tachycardia produced by 2CH3-5-HT. Furthermore, propranolol also did not block 2CH3-5-HT-induced tachycardia, indicating that an indirect neuronal effect to release norepinephrine cannot explain the increase in heart rate to 2CH3-5-HT. Tachycardia to 2CH3-5-HT (as well as to isoproterenol) was modestly inhibited, but never abolished by interruption of the autonomic nervous system with atropine or hexamethonium. 5-HT3 receptor antagonists, zacopride, ICS 205-930 and GR38032F, dose-dependently blocked the tachycardia and pressor response to 2CH3-5-HT. These data establish the presence of a 5-HT3 receptor mediating a direct positive chronotropic effect of 5-HT in conscious dogs, an effect that depends, only minimally, on the presence of an intact autonomic nervous system.

Animals↗
PubMed1990

Pharmacological and molecular evidence that the contractile response to serotonin in rat stomach fundus is not mediated by activation of the 5-hydroxytryptamine1C receptor.

The receptor mediating contraction in response to serotonin in the rat stomach fundus has not been characterized in light of the currently acceptable serotonergic receptor classification scheme. Several biochemical and pharmacological approaches to a characterization of this receptor have demonstrated nonidentity with the 5-hydroxytryptamine2 (5HT2), 5HT3, 5HT1A, and 5HT1B receptors, as defined by radiolabeled ligand binding studies in brain cortical membranes. Although there have been reports suggesting that the receptor in the rat stomach fundus may be analogous to the 5HT1C receptor, other pharmacological and biochemical studies have not been consistent with this idea. The present study utilized high affinity ligands for the 5HT1C receptor and the recently derived 5HT1C receptor cDNA clone to provide a more definitive approach to the examination of the relationship between the 5HT1C receptor and the serotonergic contractile receptor in the rat stomach fundus. Using three ligands with high affinity at the 5HT1C receptor, LY53857, ritanserin, and SCH23390, the contractile response to serotonin was inhibited by all three ligands. However, inhibition did not appear competitive nor was the inhibitory potency of these ligands consistent with their affinity at 5HT1C binding sites in brain cortical membranes. We further showed that SCH23390, unlike LY53857 and ritanserin, was also a partial agonist, producing a maximal contraction that was approximately 50% of the maximal response to serotonin in the rat stomach fundus. Thus, the use of these ligands did not support the contention that the receptor mediating serotonin-induced contractions in the rat stomach is identical to the 5HT1C receptor. In more definitive studies using a 5HT1C receptor cDNA probe, we were unable to detect hybridization of the probe with any mRNA species from the rat stomach fundus, whereas the 5HT1C receptor cDNA probe did hybridize to the 5HT1C receptor mRNA in rat brain. Because the cathepsin-D cDNA probe hybridized equally in rat brain and stomach fundus, ensuring the integrity of the RNA preparation from both tissues, the absence of measurable quantities of the 5HT1C receptor mRNA in the rat stomach was probe specific and not an artifact. Furthermore, primers specific for the rat 5HT1C receptor sequence did not detect significant levels of receptor mRNA in rat fundus, although the target sequence was amplified a minimum of 10(5)-fold in a polymerase chain reaction. These studies do not support the contention that the receptor mediating contractile responses to serotonin in the rat stomach fundus is identical to the 5HT1C receptor.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗