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D L Nelson

Publications and source records attributed to D L Nelson.

At least 199 records · Page 11Linked to original sources
PubMed1994

Targeting human IL-2 receptors for diagnosis and therapy.

The high-affinity interleukin-2 receptor (IL-2R) is a multichain receptor with at least three IL-2 binding chains: IL-2R alpha (55 kDa) bound by the monoclonal antibody anti-Tac, IL-2R beta (75 kDa) and Il-2R gamma (64 kDa). The IL-2R alpha also exists as a naturally occurring soluble molecule (sIL-2R alpha). We target the IL-2R for immune intervention since resting normal cells do not express the high-affinity IL-2R, whereas this receptor is on some cells in certain lymphoid neoplasias, select autoimmune disorders, and in individuals rejecting organ allografts. Treatments have included unmodified murine anti-Tac and radioisotopes conjugated to murine anti-Tac. Our emerging understanding of the IL-2R system continues to open possibilities for more specific immune intervention.

Animals↗
PubMed1994

Prosthesis training as a context for studying occupational forms and motoric adaptation.

OBJECTIVES: Occupational therapy authors frequently emphasize the importance of the use of objects in the development of motor skill. This study investigated the use of object-produced visual input in learning control of flexion and extension of an above-elbow training prosthesis. METHOD: Fifty-two male college students were randomly assigned to two training procedures: (a) two 1.5-min periods in which they used a flashlight attached to the hook of the prosthesis to connect dots on paper with light, or (b) the same time periods in which they had the opportunity to practice moving an equally weighted prosthesis, but without the light or dots. To assess motoric adaptation after training under one of the two conditions, each subject traced a continuous line through a maze with a pen attached to the hook. Deviations from the line were measured reliably. RESULTS: Data analysis with a Mann-Whitney test revealed that subjects in the group that trained with added materials traced with significantly more skill than subjects in the other group (one-tailed U = 225.5, p = .02). CONCLUSION: As predicted by occupational therapy theory, the object-produced visual input enhanced the learning of a motor skill relevant to rehabilitation. Although there is a need for more study across different occupations and populations, clinicians are urged to consider the possible benefits of the use of objects in the development of motor skills, as opposed to objectless exercise. Prosthetic training provides a useful context for future research addressing theoretical issues in motor learning.

Adolescent↗
PubMed1994

Pharmacological characterization of LY293284: A 5-HT1A receptor agonist with high potency and selectivity.

(-)-LY293284, (-)-4R-6-acetyl-4-(di-n-propylamino)1,3,4,5- tetrahydrobenz[c,d]indole, is a conformationally restricted tryptamine derivative with an acetyl group serving as a protophilic substitution for the hydroxyl in serotonin (5-HT). In ligand displacement studies, LY293284 had a Ki of 0.07 nM for the 5-HT1A receptor but no affinity for other monoaminergic receptors within 3 orders of magnitude. LY293284 was evaluated in in vivo models, which have been used as markers for presynaptic and postsynaptic 5-HT1A receptor activity. LY293284 decreased hypothalamic 5-hydroxyindoleacetic acid levels (ED50, 2.9 micrograms/kg s.c.) and dorsal raphe serotonergic neuron firing rate (ED50, 0.08 micrograms/kg s.c.), which are accepted indices of presynaptic activity. LY293284 also induced a reduction in body temperature in rats (ED50, 3.6 micrograms/kg s.c.), which was blocked by pretreatment with (+/-)-pindolol. Hypothermic responses of rats to 5-HT1A agonists have had both pre- and postsynaptic characteristics in previous studies. The ED50 values for 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT) in these tests were 15 to 45 times higher than those observed for LY293284. In models for postsynaptic activity, the ED50 for LY293284 for elevating serum corticosterone levels was 9.7 micrograms/kg s.c. and the minimum effective doses to induce lower lip retraction and flat posture were 3 micrograms/kg s.c. For comparison, the same indices obtained for 8-OH-DPAT were 222.4 and 100 micrograms/kg, respectively. The 5-HT syndrome responses induced by LY293284 were also attenuated by pretreatment with (+/-)-pindolol. LY293284 was 10 times more potent than 8-OH-DPAT in a drug discrimination test that used pigeons trained to identify 8-OH-DPAT. In sexual behavior tests with male rats, LY293284 induced a maximal reduction in ejaculatory latency at 0.01 micrograms/kg s.c., which was approximately 10 times higher potency than 8-OH-DPAT. In the pigeon conflict model for anxiolytic activity, LY293284 was 100 times more potent than 8-OH-DPAT in increasing punished responding. In the rat forced swim model for antidepressant-like activity, LY293284 was 30 and 35 times more potent than 8-OH-DPAT in decreasing immobility time and defecation rate. These studies have demonstrated that LY293284 is a highly selective and extremely potent 5-HT1A receptor agonist and represents a useful pharmacological tool for studying 5-HT1A receptor-mediated effects.

Animals↗
PubMed1994

Molecular cloning, functional expression, and mRNA tissue distribution of the human 5-hydroxytryptamine2B receptor.

Clones encoding a portion of the human 5-hydroxytryptamine (5-HT)2B receptor gene were isolated from a human placental genomic library. Based on distribution studies of 5-HT2B receptor mRNA, human uterus cDNA libraries were constructed and screened, resulting in the isolation of several full-length cDNA clones. These clones harbored a common single open reading frame encoding a protein of 481 amino acids. The deduced amino acid sequence of the human 5-HT2B receptor displayed 91.5% identity within the transmembrane domains and 82% identity overall with the rat 5-HT2B receptor. The human 5-HT2B receptor stably expressed in AV12-664 cells demonstrated high affinity (Kd = 10.18 +/- 1.60 nM), saturable [3H]serotonin binding, similar to that previously described for the rat 5-HT2B receptor. The pharmacological profile of the human 5-HT2B receptor was virtually identical to that of the rat 5-HT2B receptor, with the exceptions of the 5-HT2A receptor antagonists ketanserin and spiperone. Both compounds exhibited higher affinity at the human 5-HT2B receptor (ketanserin, Ki = 376 +/- 58 nM; spiperone, Ki = 697 +/- 54 nM) than at the rat 5-HT2B receptor (ketanserin, Ki = 3559 +/- 175 nM; spiperone, Ki = 3278 +/- 92 nM). Functional coupling of the human 5-HT2B receptor was also demonstrated in AV12-664 cells, where 5-HT produced a dose-dependent increase in phosphatidylinositol hydrolysis (EC50 = 27 +/- 12 nM) analogous to that seen with the rat 5-HT2B receptor. Reverse transcription-polymerase chain reaction studies revealed human 5-HT2B receptor mRNA to be expressed in many tissues, including the central nervous system. The presence of 5-HT2B receptor mRNA in human brain and not in rat brain raises the possibility that the 5-HT2B receptor may be of significance in higher brain function.

Amino Acid Sequence↗
PubMed1994

Species variations in transmembrane region V of the 5-hydroxytryptamine type 2A receptor alter the structure-activity relationship of certain ergolines and tryptamines.

Previous work has suggested that species differences in the structure-activity relationship for ergolines and tryptamines at the 5-hydroxytryptamine (5-HT)2A (formerly known as the 5-HT2) receptor are related to aliphatic substitution at the N1-position on the indole nucleus. The present work has confirmed these findings by examining the rat and human cloned 5-HT2A receptors. As previously found, N1-substitution of ergolines or tryptamines had no effect or increased affinity for the rat 5-HT2A receptor but decreased affinity for the human receptor. Also, the N1-unsubstituted analogues had higher affinity for the human 5-HT2A receptor, whereas the N1-alkyl analogues had a higher affinity for the rat receptor. By mutating the rat 5-HT2A receptor, the importance of the Ala/Ser242 species variation in amino acid sequence was examined in relation to this structure-activity relationship. Three mutations of the rat 5-HT2A receptor were made, i.e., A242S, A242V, and A242T. All three mutations resulted in functional (able to stimulate inositol phosphate hydrolysis) 5-HT2A receptors with high affinity for [3H]ketanserin and 1-(2,5-dimethoxy-4-[125I]iodophenyl)isopropylamine. The A242S mutation resulted in a pharmacological profile that was almost identical to that of the human 5-HT2A receptor but differed significantly from that of the wild-type rat receptor. This strongly suggests that the Ala/Ser242 species variation accounts for the differences in the structure-activity relationship. The A242V and A242T mutations resulted in differing but profound effects on affinity for the different ergolines and tryptamines. The results are discussed in terms of the importance of position 242 in the binding of these ligands to 5-HT2A receptors. In addition, arguments are presented that suggest that a hydrogen-bonding interaction occurs between the human 5-HT2A receptor at Ser242 and the N1-hydrogen of N1-unsubstituted ergolines and tryptamines and may serve as an important contact point in the receptor.

Amphetamines↗
PubMed1993

Identification of a family of casein kinases in Paramecium: biochemical characterization and cellular localization.

Protein phosphorylation is believed to play a role in the regulation of ciliary motility in the protozoan Paramecium tetraurelia. Five protein kinases from Paramecium, activated by cyclic nucleotides or Ca2+, have been characterized previously. We report here the identification of a family of second-messenger-independent casein kinases in Paramecium. Casein kinase activity was enriched in the soluble fraction of cilia, but there was also significant activity tightly associated with axonemes. Three ciliary casein kinase activities (soluble CKS1 and CKS2, and axonemal CKA) were separated by chromatography and characterized. The native forms of all three were monomeric, with molecular masses of 28-45 kDa as judged by in-gel kinase assays and sizing by gel filtration. CKS2 was inhibited by heparin, but CKA was unaffected and CKS1 was stimulated. All three activities preferred acidic substrates such as casein and phosvitin, but they could be distinguished by their preference for other substrates. Antibodies against mammalian casein kinase I recognized CKS1 and CKS2 in immunoblots (43 kDa), but did not stain CKA. The antibodies to casein kinase I were used to probe other cellular fractions. A 65 kDa antigen (particulate casein kinase, CKP) was enriched in particulate fractions of whole cells. This 65 kDa protein was found in isolated cell cortices, but was not present in the infraciliary lattice. This report represents the first biochemical identification of a casein kinase I family in protozoa.

Animals↗
PubMed1993

Three-dimensional quantitative structure-activity relationships of 5-HT receptor binding data for tetrahydropyridinylindole derivatives: a comparison of the Hansch and CoMFA methods.

A series of new derivatives of 3-(1,2,5,6-tetrahydropyridin-4-yl)indole (4-THPI) has been synthesized, and their dissociation constants at the 5-HT1A and 5-HT2 serotonin (5-HT) receptor subtypes have been determined. The new data were combined with similar binding data on a related set of THPI analogs reported previously (Taylor et al. Mol. Pharmacol. 1988, 34, 42-53) and used to develop 3-dimensional quantitative structure-activity relationships (3-D QSARs) for these compounds at the 5-HT1A and 5-HT2 receptor sites, by the method of comparative molecular field analysis (CoMFA). Since the previous study included several conventional QSARs obtained by Hansch analysis, and the new compounds in some cases fall within the congeneric series used in those analyses, we were able to make a direct comparison of the predictive capabilities of CoMFA and Hansch analysis using identical training and test data sets. The overall quality of actual predictions of activity by both methods appears to be about the same, as assessed by the root mean square (rms) residuals between actual and predicted pKi values. On the one hand, the compounds most poorly predicted by the Hansch analysis were 34, 35, and 37, while compounds 30-33 were relative poorly predicted by CoMFA. However, a clear advantage of CoMFA is the ability to include diversely substituted or noncongeneric analogs that must be omitted from conventional QSAR analysis. Using the entire data set of 45 THPI analogs reported here, pKi predictions for six additional compounds having 5-heteroarylindole substituents gave rms residuals of 0.46 and 0.36 for the 5-HT1A and 5-HT2 models, respectively; this is close to the experimental error of the binding data. The significance of the CoMFA field graphs in terms of molecular features required for activity and selectivity at these 5-HT receptor subtypes is discussed.

Animals↗
PubMed1993

N(1)-substituted ergolines and tryptamines show species differences for the agonist-labeled 5-HT2 receptor.

Previous studies indicated that selected ergolines and tryptamines showed species differences for affinity to the antagonist-labeled 5-HT2 receptor. The present study examined these same compounds for affinity at the agonist-labeled 5-HT2 receptor in rat and squirrel monkey cortical homogenates using [125I]DOI ([125I]1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane). As seen with the antagonist-labeled 5-HT2 receptor, N(1) alkyl substitution of either the ergolines or tryptamines resulted in a slight increase or no effect on their affinity for the agonist-labeled rat 5-HT2 receptor. In contrast, these same N(1) substitutions resulted in significant decreases in affinity for the agonist-labeled monkey 5-HT2 receptor. It was also noted that N(1)-unsubstituted ergolines and tryptamines (such as ergonovine, LY86057, LY193525 and 5-methoxytryptamine) tended to have higher affinity for the monkey versus the rat agonist-labeled receptor. However, the N(1) alkyl-substituted ergolines and tryptamines (such as mesulergine, LY53857, amesergide, N(1)-isopropyltryptamine and N(1)-isopropyl-5-methoxytryptamine) showed significantly lower affinity for the monkey versus the rat 5-HT2 receptor. These data suggest that, at least in relation to the N(1) position, ergolines and tryptamines bind in a similar orientation. These results are also discussed in terms of what amino acid differences between species may account for this structure-activity relationship.

Amphetamines↗
PubMed1993

External GTP alters the motility and elicits an oscillating membrane depolarization in Paramecium tetraurelia.

Paramecium, a unicellular ciliated protist, alters its motility in response to various stimuli. Externally added GTP transiently induced alternating forward and backward swimming interspersed with whirling at a concentration as low as 0.1 microM. ATP was 1000-fold less active, whereas CTP and UTP produced essentially no response. The response to the nonhydrolyzable GTP analogs guanosine 5'-[gamma-thio]triphosphate and guanosine 5'-[beta, gamma-imido]triphosphate was indistinguishable from that to GTP. This behavioral response was correlated with an unusual transient and oscillating membrane depolarization in both wild-type cells and the mutant pawn B, which is defective in the voltage-dependent Ca2+ current required for action potentials. This is a specific effect of external GTP on the excitability of a eukaryotic cell and, to our knowledge, is the first purinergic effect to be discovered in a microorganism.

Animals↗
PubMed1993

A second member of the novel Ca(2+)-dependent protein kinase family from Paramecium tetraurelia. Purification and characterization.

The ciliated protozoan Paramecium tetraurelia contains two protein kinase activities (CaPK-1 and CaPK-2) that are dependent on Ca2+ (Gundersen, R. E., and Nelson, D. L. (1987) J. Biol. Chem. 262, 4602-4609). We purified Ca(2+)-dependent protein kinase-1 (CaPK-1) 1,000-fold from the EGTA-extracted soluble fractions of Paramecium. The purified enzyme was a single polypeptide of 52 kDa on SDS-polyacrylamide gel electrophoresis with a native molecular mass of 60,000, suggesting that the active enzyme is a monomer. The purified kinase used casein as the best substrate in vitro, and its activity was absolutely dependent on Ca2+. The physical, catalytic and regulatory properties were clearly distinct from those of casein kinase, protein kinase C, and Ca2+/calmodulin-dependent protein kinases. CaPK-1 was half maximally activated by submicromolar (0.2 microM) free Ca2+, and the purified kinase bound Ca2+ in a blot overlay assay. CaPK-1 and the previously characterized CaPK-2 were biochemically and immunologically different enzymes sharing a similar activation mechanism. CaPK-1 and CaPK-2 appear to be members of a new family of Ca(2+)-dependent protein kinases. A protein immunologically related to the CaPKs was also detected in rat brain.

Animals↗
PubMed1993

Immunological comparison of 22S, 19S, and 12S dyneins from Paramecium cilia.

Three forms of dynein (22S, 19S, and 12S) were purified from Paramecium cilia. Two classes of monoclonal antibodies against purified 22S dynein were generated. One class reacted on immunoblots with the heavy chains of 22S, 19S, and 12S dyneins; the second class reacted with an 88 kD intermediate chain of 22S dynein. Polyclonal antiserum to the heavy chains of 22S dynein reacted with the alpha-heavy chain of 22S and 19S dyneins. A previously described antiserum raised against 22S dynein [Travis et al.: Biochim. Biophys. Acta 966:73-83, 1988] recognized the gamma-heavy chain of 22S dynein which was also present in 19S and 12S dyneins, along with the 88 and 76 kD intermediate chains of 22S dynein. This antiserum was also able to immunoprecipitate dynein from crude extracts of cilia. Electron microscopy revealed that the 22S dynein consisted mainly of two-headed particles with some three-headed particles present. The 12S dynein was mainly one-headed particles. The 19S dynein was a mixture of three-, two-, and one-headed particles. The immunological and electron microscopic studies showed that 19S dynein arises from 22S dynein, and that 12S dynein is heterogeneous, composed of the gamma-heavy chain of 22S dynein and a unique dynein ATPase. The polyclonal antibodies were also used to detect cross-reactive proteins in other organisms. Both the anti-heavy chain and the anti-22S dynein sera reacted strongly with 22S outer arm dynein of Tetrahymena, but not with the 14S dynein of this organism.

Animals↗
PubMed1993

Serum and urine soluble interleukin-2 receptor in idiopathic nephrotic syndrome.

Although a cellular immune pathogenesis is suspected in idiopathic nephrotic syndrome of childhood (INS), there is scant direct evidence of in vivo immune activation. In order to investigate cytokine cascade activation in INS, soluble interleukin-2 receptor (sIL-2R) in plasma and urine was characterized and its levels measured in INS patients during relapse. Immunochemically detectable sIL-2R had a molecular mass of 35-46 kDa in both serum and urine and the molecule appears to be excreted intact; the pI was 5.05. INS patients had elevated serum sIL-2R levels compared with adult normal controls (845 +/- 97 vs. 373 +/- 47 U/ml, P = 0.001) and were significantly higher than previously published age-matched controls. Urinary excretion of sIL-2R was 47.2 +/- 13 U/mg creatinine in patients. Both the sIL-2R excretion rate and the fractional excretion of sIL-2R were positively correlated with the excretion of albumin (P = 0.02 and 0.002, respectively). These increased serum and urine levels occurred whether relapse was or was not associated with an intercurrent illness. We conclude that: (1) despite increased sIL-2R excretion during INS relapse, serum levels are significantly elevated; (2) while the elevated urinary levels could result from enhanced intrarenal production, they more likely reflect the increased serum levels; (3) the elevated sIL-2R levels support an immune pathogenesis in INS.

Adolescent↗
PubMed1993

Increased serum soluble interleukin-2 receptors in schizophrenic monozygotic twins.

There is a confusing history of immune findings associated with schizophrenia. At least some of these discrepant results may be artifacts caused by heterogeneity. In an attempt to decrease heterogeneity, we studied three groups of monozygotic twins who were either discordant for schizophrenia, concordant and ill, or concordant and well. This comparison minimizes environmental and genetic variance, and heightens differences that are actually due to the disorder. Overall, schizophrenic subjects had higher levels of serum soluble interleukin-2 receptors (SIL-2Rs) than unaffected individuals (480.8, SD 238.6 U/ml vs 380.9, SD 170.6 U/ml; F = 5.256, df = 1.61, P = 0.02). When data from discordant and concordant twin groups were analysed separately, both the discordant ill twins (P = 0.06) and concordant ill twin pairs (P = 0.08) showed trends towards higher serum SIL-2R levels than their respective control groups. These data contribute to the growing body of evidence that immune activation is associated with some forms of schizophrenia.

Adult↗
PubMed1993

Lunotriquetral arthrodesis.

Lunotriquetral arthrodesis has been recommended for the treatment of disabling pain at the lunotriquetral joint after more conservative measures have failed. We retrospectively analyzed a series of 22 patients treated with a lunotriquetral arthrodesis for presence of fusion, method of fixation, duration of immobilization, carpal alignment, pain relief, and ability to return to work. Fixation with a Herbert screw supplemented with a Kirschner wire (K-wire) was superior to fixation with K-wires alone. Immobilization longer than 6 weeks was superior to immobilization less than 6 weeks. Combined use of a Herbert screw supplemented with a K-wire and immobilization longer than 6 weeks resulted in union in all patients, even when performed for nonunion after a prior attempted arthrodesis. Pain was improved in all patients; all patients who previously were working returned to work. Routine posteroanterior and lateral x-ray films often failed to adequately profile the arthrodesis site. We recommend using a Herbert screw supplemented with a K-wire for lunotriquetral arthrodesis and keeping the patient in a cast until fusion is documented, usually at least 8 weeks. Fluoroscopic spot views or tomograms are recommended to demonstrate bone fusion.

Adult↗
PubMed1993

Trinucleotide repeat expansions in neurological disease.

During the past year, new examples of human neurological disease have been discovered that have an unprecedented type of mutation as their cause: the remarkable expansion of trinucleotide repeats. These triplet repeats are normally polymorphic and exonic, though not always coding. In disease states they become markedly unstable and may expand moderately or by thousands of repeats in a single generation, influencing gene expression, message stability or protein structure.

Fragile X Syndrome↗
PubMed1993

Methods for finding genes. A major rate-limiting step in positional cloning.

Identification of transcribed sequences from within genomic regions has been a major rate-limiting step in the pursuit of genes involved in many human genetic diseases. Early efforts focused primarily on screening of cDNA libraries, identification of evolutionarily conserved sequences, and northern blot hybridization. In recent years, several innovative techniques for gene identification have been devised. These techniques expand the size of the genomic region capable of being scanned for genes, while also allowing detection of genes regardless of their expression patterns. This article reviews several new and older techniques and discusses the advantages and limitations of each.

Cloning, Molecular↗