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Biomedical subjects

L Isaac

Publications and source records attributed to L Isaac.

At least 37 records · Page 2Linked to original sources
PubMed1994

Pharmacological characterization of dynorphin A (1-17)-induced effects on spinal cord-evoked potentials.

Dynorphin A (1-17) was applied directly onto the spinal cord of rats during electrophysiologic recording of the dorsal root potential (DRP) and the ventral root potentials (VRPs), i.e., monosynaptic reflex and polysynaptic reflexes. Dynorphin application resulted in a dose-dependent depression of the DRP (ED50, 4.5 nmol) which persisted for 30 to 50 min. This effect was not antagonized by nor-binaltorphimine, a kappa-opioid receptor antagonist. During this depression we observed a potentiation of the VRPs which persisted for 4 to 5 min and preceded depression of the VRPs (ED50, 4.0-4.9 nmol). The depression of the VRPs was antagonized competitively by nor-binaltorphimine, although the potentiation was not. beta-Funaltrexamine, a mu-opioid receptor antagonist, had no influence on dynorphin-induced changes of evoked potentials. These data indicate that dynorphin-induced depression of the VRPs is mediated by kappa-opioid receptor activity, whereas neither potentiation of the VRPs nor depression of the DRP appears to be mediated by an opioid receptor effect.

Animals↗
PubMed1993

Dynorphin A (1-13) potentiates dynorphin A (1-17) on loss of the tail-flick reflex after intrathecal injection in the rat.

Dynorphin A (1-13) or dynorphin A (1-17) administered intrathecally to rats induces a dose-dependent loss of the tail-flick reflex and a reversible hindlimb paralysis. Although their potency is comparable, dynorphin A (1-17) is less efficacious in producing loss of the tail-flick reflex. These data suggest that dynorphin A (1-17) acts as a partial agonist or exerts its neurotoxic effect in the presence of an endogenous non-competitive inhibitor. To determine whether these substances act at similar sites we performed isobolographic analysis. This analysis revealed that dose-additivity exists for paralysis whereas deviation from dose-additivity exists for loss of the tail-flick reflex.

Analgesics↗
PubMed1992

Effect of electroconvulsive shock on 5-HT2 and alpha 1-adrenoceptors and phosphoinositide signalling system in rat brain.

We studied the effect of repeated administration of electroconvulsive shock (ECS) on alpha 1-adrenoceptor subtype (alpha 1A and alpha 1B) and 5-HT2 (serotonin-2) receptors and receptor-mediated phosphoinositide (PI) hydrolysis in rat cerebral cortex. We observed that repeated administration with ECS significantly increased the density of 5-HT2 receptors, as labeled by [3H]ketanserin, as well as 5-HT-stimulated [3H]inositol-1-phosphate ([3H]IP1) in rat cerebral cortex. We also observed that repeated ECS administration caused a significant increase in the number of alpha 1-adrenoceptors and the alpha 1B-adrenoceptor subtype as measured by (+/-)-beta-([125I]iodo-4-hydroxyphenyl)-ethyl-aminomethyl-tetralone binding. However, it had no significant effects on norepinephrine (NE)-stimulated [3H]IP1 formation or alpha 1A-adrenoceptor subtype. These results thus suggest that up-regulation of 5-HT2 receptors after administration with ECS is associated with increased 5-HT-stimulated [3H]IP1 formation. The lack of effects on NE-stimulated PI turnover in ECS treated rats may be due to its lack of effect on the alpha 1A-adrenoceptor subtype.

Animals↗
PubMed1992

Structural requirements for thioester bond formation in human complement component C3. Reassessment of the role of thioester bond integrity on the conformation of C3.

A unique thioester bond, formed between the side chains of neighboring C and Q residues, is present in complement components C3 and C4 and the protease inhibitor alpha 2-macroglobulin. This structure is essential for mediating covalent attachment to target acceptors and also for maintaining these proteins in their native conformation. An examination of the residues in the immediate vicinity of the C and Q reveals a very high degree of sequence similarity among the three proteins which crosses species barriers. The following is the sequence flanking the thioester residues in C3, the highly conserved amino acids being underlined and the the thioester-forming residues being indicated by italics: 1005V-T-P-S-G-C-G-E-Q-N-M-I-G-M-T-P-T1021. Through a site-directed mutagenesis and cDNA expression approach, we have examined the importance of the conserved amino acids in the formation, stability, and function of the thioester bond in C3. The behavior of the mutants fell into three categories. The potential loss in peptide backbone flexibility by the replacement of G1009 by A or S was permissive to thioester formation and function as was replacement of M1015 by the still fairly bulky residue F. In contrast, replacement of M1015 by A resulted in an alpha-chain which was highly unstable toward proteolytic degradation. The third category, which included mutant molecules P1007G, P1020G, E1012Q, and Q1013N, displayed an unusual phenotype in which both the autolytic fragmentation and the hemolytic activity characteristics of thioester-intact molecules were absent. However, like their wildtype counterpart, these molecules retained the ability to be cleaved by C3 convertase (C4b2a), a conformation-dependent property that is normally lost in the conversion of native C3 to thioester-hydrolyzed C3(H2O). Since an identical functional profile was obtained when the thioester was deliberately prevented from forming in the mutant C1010A, we conclude that if a stable thioester fails to form during biosynthesis, at least parts of the mature protein can adopt a more native-like conformation than is the case when the thioester is first formed and then hydrolyzed in the mature protein. In view of these new findings, the interpretation of the previously observed correlation between the loss of thioester integrity and the adoption of a C3b-like conformation must be reassessed.

Amino Acid Sequence↗
PubMed1992

Putative neurotoxicity of SKF 38393 and other D1 dopaminergic drugs investigated in rat striatum.

The recently alleged neurotoxicity of the D1 receptor agonist, SKF 38393, was investigated in rat striatum by measuring the enzymes acetylcholinesterase (AChE) and glutamate decarboxylase (GAD). First, unilateral intrastriatal microinjection of the excitotoxin kainic acid (2 micrograms in 1 microliter) was shown to evoke vigorous contraversive circling, followed 1 or 2 weeks later by profound decreases in striatal AChE (24 and 54%), GAD (51 and 75%), and protein (36 and 47%), as well as loss of GAD (45% at 2 weeks) in the ipsilateral substantia nigra. Similar striatal treatments with SKF 38393 (30 micrograms in 0.5-1 microliter), the related benzazepines SKF 82526 (D1 agonist, 30 micrograms in 1 microliter) and SCH 23390 (D1 antagonist, 5 micrograms in 1 microliter), or the phenanthridine D1 agonist CY 208-243 (5 micrograms in 1 microliter) failed to affect the rats' behaviour or their striatal levels of AChE, GAD, and protein. Intrastriatal SKF 38393 (30 micrograms in 0.5 microliter) also had no influence on these enzymes in the substantia nigra. It is concluded that none of the D1 dopaminergic compounds examined here was neurotoxic toward the many different cell groups that contain AChE and/or GAD in the striatum.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗
PubMed1991

Similar effects of treatment with desipramine and electroconvulsive shock on 5-hydroxytryptamine1A receptors in rat brain.

The effect of chronic and acute treatment with desipramine (DMI) and electroconvulsive shock (ECS) on 5-hydroxytryptamine1A (5-HT1A) receptors was determined in the cortex and the hippocampus brain regions of rats. We observed that chronic treatment with both DMI and ECS significantly decreased 5-HT1A receptors, as determined by [3H]8-hydroxy-2-(di-n-propylamino)tetralin [( 3H]8-OH-DPAT) binding, in the cortex but not in the hippocampus. Acute treatment with DMI or ECS did not significantly alter the 5-HT1A receptors in the cortex. Neither chronic nor acute treatment influenced KD of [3H]8-OH-DPAT binding in the cortex or hippocampus. These results thus suggest that in contrast to the effects on 5-HT2 receptors, tricyclics as well as ECS produce similar effects on 5-HT1A receptors, suggesting that this site may represent a common site of action for antidepressant treatment.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1991

A strychnine-sensitive site is involved in dynorphin-induced paralysis and loss of the tail-flick reflex.

Dynorphin A(1-13) administered intrathecally to rats induces a reversible hindlimb paralysis and permanent loss of the tail-flick reflex in a dose-dependent and all-or-none manner. The loss of the tail-flick reflex has been determined to result from neurotoxicity linked to the N-methyl-D-aspartate (NMDA) receptor. Recently, it has been reported that NMDA antagonists attenuate irreversible paralysis induced by dynorphin A(1-17) and dynorphin A(2-17). In the present studies, we examined whether repeated injections of dynorphin A(1-13) acetate salt could change the characteristics of the reversible paralysis. Injections repeated every 48 h resulted in hindlimb paralysis upon each injection which was not different in terms of magnitude or duration (P greater than 0.60). Injections repeated at 2 h intervals resulted in desensitization of the paralytic effects (P less than 0.05). We also examined if strychnine sulfate, a glycine antagonist would alter the paralytic response to dynorphin. Strychnine protected rats from paralysis (P less than 0.01) and loss of the tail-flick reflex with an ED50 of 7 nmol. We conclude that the reversible paralysis induced by dynorphin A(1-13) is repeatable which suggests that the paralysis results from nontoxic or subtoxic actions of dynorphin. Desensitization to the paralytic effects occurs with closely spaced injections by some unknown mechanism. In addition, we conclude that the spinal glycinergic inhibitory system may participate in the induction of the paralysis because strychnine antagonizes dynorphin-induced paralysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Complexes of .NO with nucleophiles as agents for the controlled biological release of nitric oxide. Vasorelaxant effects.

Selected nucleophile/nitric oxide adducts [compounds which contain the anionic moiety, XN(O-)N = O] were studied for their ability to release nitric oxide spontaneously in aqueous solution and for possible vasoactivity. The diversity of structures chosen included those in which the nucleophile residue, X, was that of a secondary amine [Et2N, as in [Et2NN(N = O)O]Na, 1], a primary amine [iPrHN, as in [iPrHNN(N = O)O]Na, 2], a polyamine, spermine [as in the zwitterion H2N(CH2)3NH2+(CH2)4N[N(N = O)O-](CH2)3NH2, 3], oxide [as in Na[ON(N = O)O]Na, 4], and sulfite [as in NH4[O3SN(N = O)O]NH4, 5]. The rate constants (k) for decomposition in pH 7.4 phosphate buffer at 37 degrees C, as measured by following loss of chromophore at 230-260 nm, were as follows: 1, 5.4 x 10(-3) s-1; 2, 5.1 x 10(-3) s-1; 3, 0.30 x 10(-3) s-1; 4, 5.0 x 10(-3) s-1; and 5, 1.7 x 10(-3) s-1. The corresponding extents of nitric oxide release (ENO) were 1.5, 0.73, 1.9, 0.54, and 0.001 mol/mol of starting material consumed, respectively, as determined from the integrated chemiluminescence response. Vasodilatory activities expressed as the concentrations required to induce 50% relaxation in norepinephrine-constricted aortic rings bathed in pH 7.4 buffer at 37 degrees C (EC50) were as follows: 1, 0.19 microM; 2, 0.45 microM; 3, 6.2 microM; 4, 0.59 microM; and 5, 62 microM. Vasorelaxant potency (expressed as 1/EC50) was strongly correlated with the quantity of .NO calculated from the physicochemical data to be released in the interval required to achieve maximum relaxation at the EC50 doses (r = 0.995). This suggests that such nucleophile/.NO adducts might generally be useful as vehicles for the nonenzymatic generation of nitric oxide, in predictable amounts and at predictable rates, for biological purposes. The particular significance for possible drug design is underscored in the very favorable potency comparison between several of these agents and the established nitrovasodilators sodium nitroprusside and glyceryl trinitrate (EC50 values of 2.0 and greater than 10 microM, respectively) in parallel aortic ring tests.

Animals↗
PubMed1991

Dynorphin-induced depression of the dorsal root potential in rat spinal cord: a possible mechanism for potentiation of the C-fiber reflex.

Dynorphin A (1-13) acetate salt was applied to the exposed spinal cord of rats during electrophysiological recording of the dorsal root potential (DRP) and the DR III arising from dorsal root stimulation. Simultaneously, ventral root potentials indicative of monosynaptic and polysynaptic myelinated pathways were recorded. Amplitudes of the DRP and DR III were decreased for 30 to 60 min in a dose-dependent manner after dynorphin administration with ED50 values of 21 and 28 nmol, respectively. Amplitude of ventral root potentials was also decreased with a maximal effect at 5 min postdrug treatment. In contrast, we (Caudle and Isaac, J. Pharmacol. Exp. Ther. 246: 508-513, 1988b) showed that the polysynaptic unmyelinated pathway (C-fibers) was greatly enhanced by dynorphin treatment and that this pathway was the locus of excitotoxicity (Caudle and Isaac, Brain Res. 443: 329-332, 1988a). Because the DRP reflects gamma-aminobutyric acid-mediated presynaptic inhibition of primary afferent terminals, these data indicate that dynorphin suppresses gamma-aminobutyric acid-mediated inhibition thereby disinhibiting primary afferent transmission. This presynaptic disinhibition may be the mechanism for the selective potentiation of the C-fiber pathway which may account for the selective neuron toxicity, i.e., loss of the C-fiber pathway and sparing of the monosynaptic and polysynaptic myelinated pathways after dynorphin administration.

Action Potentials↗
PubMed1990

MK-801 blocks dynorphin A (1-13)-induced loss of the tail-flick reflex in the rat.

Dynorphin A (1-13) administered intrathecally to rats results in a dose-dependent loss of the tail-flick reflex. This effect is mediated, at least in part, by N-methyl-D-aspartate receptors. We examined the influence of pretreatment or post-treatment with MK-801 on this behavioral response. MK-801 administered i.p. 30 min prior to dynorphin provided dose-dependent protection against loss of the tail-flick reflex with an ED50 of 0.06 mg/kg. MK-801 administered after dynorphin had a dose- and time-dependent protective action. The dose of 0.06 mg/kg protected 63% of the animals from loss of the tail-flick reflex when injected 15 min after dynorphin. In contrast, 3 mg/kg did not protect animals when injected 15 min after dynorphin, but did protect 50% of the animals when injected 30 min post-dynorphin. Although we cannot exclude other effects mediated by MK-801, these data support our previous findings that dynorphin-induced loss of the tail-flick reflex involves the N-methyl-D-aspartate-receptor complex and support the contention that the process(es) initiated by dynorphin injection proceed rapidly (minutes rather than hours).

Animals↗
PubMed1990

Lytic, agglutinating, and opsonizing effect of alpha 2-macroglobulin on sheep red blood cells.

Mouse alpha 2-macroglobulin (alpha 2M) induced agglutination and lysis of sheep red blood cells depending on temperature and time of incubation in vitro. When these erythrocytes (E) were treated with a subagglutinanting dose of alpha 2M, they were adhered to and phagocytosed by thioglycollate-elicited and BCG-activated mouse peritoneal macrophages. Phagocytosis was not observed when resident peritoneal macrophages were tested. alpha 2M also was able to dissociate sheep red blood cells previously aggregated by IgM anti-E.

Animals↗
PubMed1990

Trypanosoma cruzi: killing and enhanced uptake by resident peritoneal macrophages treated with alpha-2-macroglobulin.

We report that alpha-2-macroglobulin (A2M), the physiologically important plasma protease inhibitor and suspected immunomodulator, alters the functional ability of murine resident peritoneal macrophages (RM) to ingest and kill the infective trypomastigote stage of Trypanosoma cruzi, the aetiological agent of Chagas' disease. Treatment of RM with 500 micrograms/ml A2M for 30 min enhanced the uptake of trypomastigotes, epimastigotes, and amastigotes by 125%, 46%, and 300%, respectively. The same treatment also increased the phagocytosis of sheep erythrocytes opsonized with complement and IgG as well as of galactosylated asialoerythrocytes. After 60-90 min parasite-cell interaction, epi- and amastigotes were killed by the RM, whereas the infection with trypomastigotes was controlled only after 24 h. Other protease inhibitors, bovine serum albumin, and LPS showed no such effect. The production of hydrogen peroxide was not affected by A2M treatment, but the ultrastructural aspects showed trypomastigote damage and enhancement of macrophage membrane ruffling, indicative of macrophage activation. These results suggest that A2M has the ability to modulate, at least functionally, certain receptor-mediated endocytic pathways that, in concert with an activation of possibly oxygen-independent microbicidal mechanisms, could contribute to resistance against the parasite.

Animals↗
PubMed1989

Localization of dynorphin-induced neurotoxicity in rat spinal cord.

Intrathecally injected dynorphin A (1-13) in rats results in a reversible hindlimb paralysis and an irreversible loss of the tail-flick reflex. Histologic examination of the spinal cords of dynorphin treated rats demonstrated dead and/or dying neurons predominately localized in the central area which approximates Rexed lamina VII and X. In this area a maximum effect of the dynorphin-induced neurotoxicity is evident. Thus, the dynorphin-induced neuron death is suggestive of an anatomical selectivity.

Animals↗
PubMed1988

A novel interaction between dynorphin(1-13) and an N-methyl-D-aspartate site.

Dynorphin injected intrathecally in the rat results in a neurotoxicity behaviorally expressed as an irreversible loss of the thermally evoked tail-flick reflex. The excitatory amino acid antagonists DL-2-amino-5-phosphonovalerate (APV) and gamma-D-glutamylglycine (DGG) blocked the loss of the tail-flick reflex. The order of potency (APV greater than DGG) suggests that the N-methyl-D-aspartate (NMDA) subclass of excitatory amino acid receptors participate in the neurotoxicity. Additionally, intrathecal injection of APV results in a reversible loss of the tail-flick reflex, whereas with DGG doses which block the tail-flick reflex also result in hindlimb paralysis. These data suggest that neurotransmission in the tail-flick reflex pathway is, in part, mediated by NMDA receptors. From these and previous findings it was concluded that dynorphin neurotoxicity results from enhanced, excitotoxic, transmission across these synapses utilizing NMDA receptors.

2-Amino-5-phosphonovalerate↗
PubMed1988

In vitro models for testing the metabolic effects of myelographic contrast media.

Water-soluble nonionic x-ray contrast media have greatly improved the quality and safety of myelography. Toxic side effects are still observed however. The side effects are generally worse with the first nonionic agent, metrizamide, which has a glucoselike side group. Two in vitro models were developed to examine the effects of contrast media on glucose metabolism. Using rat hippocampus slices, the authors observed significant depression of carbon dioxide production by metrizamide and by deoxyglucose, a known metabolic inhibitor. Iohexol and iopamidol did not cause significant depressions. In rat brain synaptosomes the authors did not observe a depression of the uptake of deoxyglucose 14C by any media tested. These studies indicate that metrizamide can create metabolic depression but that it does not compete with glucose for the membrane glucose carrier.

Animals↗