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W Shen

Publications and source records attributed to W Shen.

At least 19 recordsLinked to original sources
PubMed1999

A seven-transmembrane, G protein-coupled receptor, FPRL1, mediates the chemotactic activity of serum amyloid A for human phagocytic cells.

We have previously reported (Badolato, R., J.M. Wang, W.J. Murphy, A. R. Lloyd, D.F. Michiel, L.L. Bausserman, D.J. Kelvin, and J.J. Oppenheim. 1994. J. Exp. Med. 180:203; Xu, L., R. Badolato, W.J. Murphy, D.L. Longo, M. Anver, S. Hale, J.J. Oppenheim, and J.M. Wang. 1995. J. Immunol. 155:1184.) that the acute phase protein serum amyloid A (SAA) is a potent chemoattractant for human leukocytes in vitro and mouse phagocytes in vivo. To identify the signaling mechanisms, we evaluated patterns of cross-desensitization between SAA and other leukocyte chemoattractants. We found that the chemotactic bacterial peptide, N-formyl- methionyl-leucyl-phenylalanine (fMLP), was able to specifically attenuate Ca2+ mobilization in human phagocytes induced by SAA, but only at very high concentrations, suggesting that SAA uses a low affinity fMLP receptor. Here we demonstrate that SAA selectively induced Ca2+ mobilization and migration of HEK cells expressing FPRL1, a human seven-transmembrane domain phagocyte receptor with low affinity for fMLP, and high affinity for lipoxin A4. Furthermore, radiolabeled SAA specifically bound to human phagocytes and FPRL1-transfected 293 cells. In contrast, SAA was not a ligand or agonist for FPR, the high affinity fMLP receptor. Thus, SAA is the first chemotactic ligand identified for FPRL1. Our results suggest that FPRL1 mediates phagocyte migration in response to SAA.

Amino Acid Sequence↗
PubMed1998

Effect of nitrous oxide on excitatory and inhibitory synaptic transmission in hippocampal cultures.

Nitrous oxide (N2O; laughing gas) has been a widely used anesthetic/analgesic since the 19th century, although its cellular mechanism of action is not understood. Here we characterize the effects of N2O on excitatory and inhibitory synaptic transmission in microcultures of rat hippocampal neurons, a preparation in which anesthetic effects on monosynaptic communication can be examined in a setting free of polysynaptic network variables. Eighty percent N2O occludes peak NMDA receptor-mediated (NMDAR) excitatory autaptic currents (EACs) with no effect on the NMDAR EAC decay time course. N2O also mildly depresses AMPA receptor-mediated (AMPAR) EACs. We find that N2O inhibits both NMDA and non-NMDA receptor-mediated responses to exogenous agonist. The postsynaptic blockade of NMDA receptors exhibits slight apparent voltage dependence, whereas the blockade of AMPA receptors is not voltage dependent. Although the degree of ketamine and Mg2+ blockade of NMDA-induced responses is dependent on permeant ion concentration, the degree of N2O blockade is not. We also observe a slight and variable prolongation of GABAA receptor-mediated (GABAR) postsynaptic currents likely caused by previously reported effects of N2O on GABAA receptors. Despite the effects of N2O on both NMDA and non-NMDA ionotropic receptors, glial glutamate transporter currents and metabotropic glutamate receptor-mediated synaptic depression are not affected. Paired-pulse depression, the frequency of spontaneous miniature excitatory synaptic currents, and high-voltage-activated calcium currents are not affected by N2O. Our results suggest that the effects of N2O on synaptic transmission are confined to postsynaptic targets.

Animals↗
PubMed1998

Metabotropic and ionotropic glutamate receptors regulate calcium channel currents in salamander retinal ganglion cells.

1. Glutamate suppressed high-voltage-activated barium currents (IBa, HVA) in tiger salamander retinal ganglion cells. Both ionotropic (iGluR) and metabotropic (mGluR) receptors contributed to this calcium channel inhibition. 2. Trans-ACPD (1-aminocyclopentane-trans-1S,3R-dicarboxylic acid), a broad-spectrum metabotropic glutamate receptor agonist, suppressed a dihydropyridine-sensitive barium current. Kainate, an ionotropic glutamate receptor agonist, reduced an omega-conotoxin GVIA-sensitive current. 3. The relative effectiveness of selective agonists indicated that the predominant metabotropic receptor was the L-2-amino-4-phosphonobutyrate (L-AP4)-sensitive, group III receptor. This receptor reversed the action of forskolin, but this was not responsible for calcium channel suppression. l-AP4 raised internal calcium concentration. Antagonists of phospholipase C, inositol trisphosphate (IP3) receptors and ryanodine receptors inhibited the action of metabotropic agonists, indicating that group III receptor transduction was linked to this pathway. 4. The action of kainate was partially suppressed by BAPTA, by calmodulin antagonists and by blockers of calmodulin-dependent phosphatase. Suppression by kainate of the calcium channel current was more rapid when calcium was the charge carrier, instead of barium. The results indicate that calcium influx through kainate-sensitive glutamate receptors can activate calmodulin, which stimulates phosphatases that may directly suppress voltage-sensitive calcium channels. 5. Thus, ionotropic and metabotropic glutamate receptors inhibit distinct calcium channels. They could act synergistically, since both increase internal calcium. These pathways provide negative feedback that can reduce calcium influx when ganglion cells are depolarized.

Ambystoma↗
PubMed1998

Amino acid residues in Std1 protein required for induction of SUC2 transcription are also required for suppression of TBPDelta57 growth defect in Saccharomyces cerevisiae.

The STD1 gene of Saccharomyces cerevisiae was isolated independently as a multicopy suppressor of a dominant negative mutation in the TATA-binding protein and of a mutation in the Snf1/Snf4 kinase complex, suggesting that Std1 might couple the Snf1 kinase signaling pathway to the transcriptional machinery. In order to identify the protein domains that specify these activities of the Std1 protein, a plasmid library of randomly mutagenized STD1 genes was screened for loss of function alleles using complementation of the raffinose growth defect of a std1-, mth1- strain as an assay. One missense allele (P236S) with complete loss of function at 30 degreesC and four missense alleles (L173F, E225K, S269L and E274K) that conferred a temperature sensitive phenotype were identified. The C-terminal 20 residues of Std1 were essential for SUC2 derepression, whereas the deletion of the N-terminal 96 residues did not affect SUC2 gene induction. Std1 mutants that lost the ability to induce SUC2, were also unable to suppress the growth defect caused by the expression of the dominant negative TBPDelta57 protein, suggesting that these two genetic screens may be detecting the same biological activity.

Alleles↗
PubMed1998

Biodegradation in vitro and retention in the rabbit eye of crosslinked poly(1-vinyl-2-pyrrolidinone) hydrogel as a vitreous substitute.

To elucidate the relatively short retention of crosslinked poly(1-vinyl-2-pyrrolidinone) hydrogels in the eye when used as potential vitreous substitutes, a 14C-labeled hydrogel was produced and subjected to both in vitro biodegradation assays and in vivo experiments. The polymer was synthesized by the free-radical copolymerization of 99% 1-vinyl-2-pyrrolidinone with 1% 14C-methyl methacrylate in the presence of ethylene glycol dimethacrylate (0.1%) as crosslinking agent. The in vitro protocol for assessing the biodegradation included the incubation of hydrogel with hydrolases (trypsin or collagenase), followed by examination of changes in its physical characteristics and by monitoring its residual radioactivity, as well as by detection of possible degradation products. Within the maximum duration of experiments (4 weeks), none of the procedures indicated biodegradation of polymer. The hydrogel was also injected into the vitreous humor of rabbits and followed up to 4 weeks. Residual radioactivity measurements of the vitreous contents indicated that 50% of the polymer was removed by the end of this period. Histopathologic examination revealed cell infiltrates of the mononuclear phagocyte system in both vitreous and retinal tissue. A possible phagocyte-mediated mechanism for the dissipation of hydrogel is discussed.

Animals↗
PubMed1998

Production and testing of Schistosoma japonicum candidate vaccine antigens in the natural ovine host.

The objectives of this work were to clone and express Chinese strain Schistosoma japonicum antigens and evaluate their immunogenicity and protective efficacy in the natural ovine host in China. Recombinant antigens selected for testing were: isoforms of glutathione S-transferase Sj28GST and Sj26GST; the large hydrophilic domain of Sj23, the homologue of the protective S. mansoni membrane antigen Sm23; and a 3' fragment of S. japonicum paramyosin. In addition, Chinese strain S. japonicum native paramyosin and GST were purified and used for vaccination. Antigens were co-administered with Freund's adjuvants or BCG. We also examined the effects of co-administration of native unfractionated GSTs with keyhole limpet haemocyanin (KLH), which shares a cross-reactive protective epitope with schistosomes. These are the first side-by-side comparisons of candidate defined-antigen schistosomiasis vaccines in a natural host. Significant partial protection was obtained with each of the antigens tested. Less protection was obtained with a recombinant fragment of S. japonicum paramyosin compared with native paramyosin. Co-administration of native GST and KLH was no more effective than vaccination with either antigen alone. Although encouraging levels of protection against S. japonicum were demonstrated using each of these antigens, further work is needed to optimise vaccine delivery and vaccination schedules.

Adjuvants, Immunologic↗
PubMed1998

Lack of desensitization and enhanced efficiency of calcium channel promoter in conscious dogs with heart failure.

The goal of this study was to compare responses to a calcium promoter, BAY y 5959, and dobutamine (Dob) in heart failure (HF). Dogs (n = 9) were chronically instrumented and studied in the conscious state before and after pacing-induced HF. In the control state, BAY y 5959 (20 microgram. kg-1. min-1) increased the first derivative of left ventricular (LV) pressure (dP/dt) by 83 +/- 8% and mean arterial pressure (MAP) by 8 +/- 2% and decreased heart rate (HR) by 30 +/- 3%. With Dob (10 microgram. kg-1. min-1) LV dP/dt rose similarly (+80 +/- 6%), but HR also rose (+25 +/- 4%) (P < 0.05 vs. BAY y 5959). After HF developed, BAY y 5959 still increased LV dP/dt by 108 +/- 8% and MAP by 21 +/- 2% and decreased HR by 28 +/- 4%, whereas Dob increased LV dP/dt by only 50 +/- 7% (P < 0.05 vs. BAY y 5959) and MAP by 7 +/- 3%, and HR did not change (+3 +/- 3%) (P < 0.05 vs. BAY y 5959). In HF, cardiac work increased more (P < 0. 05) with BAY y 5959 (+105 +/- 13%) compared with Dob (+47 +/- 11%), yet myocardial oxygen consumption increased similarly with the two drugs. Accordingly, mechanical efficiency increased more (P < 0.05) with BAY y 5959 (+73 +/- 14%) than with Dob (+17 +/- 12%). These data indicate that 1) increases in contractility mediated directly by Ca2+ are relatively resistant to desensitization in HF; and 2) the calcium-channel promoter can produce increases in myocardial contractility and cardiac work similar to those of Dob at a significantly lower oxygen cost, thereby enhancing mechanical efficiency in HF.

Animals↗
PubMed1997

Sestamibi scanning is inadequate for directing unilateral neck exploration for first-time parathyroidectomy.

OBJECTIVE: To determine whether technetium Tc 99m sestamibi scanning is accurate enough to allow surgeons to perform unilateral neck exploration for first-time parathyroidectomy in patients with primary hyperparathyroidism. DESIGN: Retrospective review. SETTING: University tertiary care center. PATIENTS: Forty patients with primary hyperparathyroidism who underwent sestamibi scanning before first-time parathyroidectomy, of whom 28 had single adenomas, 9 had multiple adenomas, and 3 had hyperplasia. INTERVENTIONS: All 40 patients underwent bilateral neck exploration with identification of 4 parathyroid glands. MAIN OUTCOME MEASURES: We compared the results of preoperative sestamibi scanning with operative and histologic findings. We then used these data to calculate the projected success rates of parathyroidectomy if unilateral neck explorations had been performed based on the results of sestamibi scanning, instead of bilateral explorations. RESULTS: Sestamibi scanning was correct in 20 (71%) of 28 patients with single adenomas, 4 (44%) of 9 patients with multiple adenomas, and 0 (0%) of 3 patients with hyperplasia. If unilateral neck explorations had been performed on the basis of localization by sestamibi scanning, parathyroidectomy would have failed in 4 (10%) of 40 patients. CONCLUSIONS: Sestamibi scanning, although helpful, is inadequate for directing unilateral neck exploration for first-time parathyroidectomy. Surgeons who perform unilateral neck exploration based on the results of sestamibi scanning will record a higher failure rate and incur higher costs than those who perform bilateral neck exploration for first-time parathyroidectomy.

Adolescent↗
PubMed1997

Mechanism of the hardening process for a hydroxyapatite cement.

The dynamics for the hydraulic process of calcium phosphate cement (CPC) were investigated by X-ray diffraction quantitative analysis. The results show that the hardening process of CPC is initially controlled by the dissolution of reactants in a 4-h period and subsequently by diffusion through the product layer of hydroxyapatite (HAP) around the grains. The compressive strength rises approximately linearly with the increase of the extent of conversion in a 4-h period, and a maximum compressive strength of about 51 MPa, which is superior to those reported by the references, is obtained in 4 h. Then the compressive strength drops a little with an increase in the extent of conversion. The final product of setting reaction is acicular HAP crystal. Crystal seed not only reduces the setting time but also drops the compressive strength. The variation of pH in CPC slurry from 7.5 to 10.5 reveals that the control step of the dissolution process in the hardening process is the dissolution of dicalcium phosphate anhydrous and the presence of crystal seed will reduce the supersaturation to produce HAP.

Biocompatible Materials↗
PubMed1997

Evaluation of the biocompatibility of a nonceramic hydroxyapatite.

In this study, the biocompatibility of calcium phosphate cement (CPC) was systematically investigated, which included systemic injection acute toxicity assay, cell culture cytotoxicity assay, gene mutation assay (Ames test), chromosome aberration assay (micronucleus test), DNA damage assay (unscheduled DNA synthesis test) and implant histological evaluation. The results showed that the CPC had no toxicity and all tests for mutagenicity and potential carcinogenicity of CPC extracts are negative. CPC was implanted into the femurs of rabbits under no-load condition and the histology of specimens revealed that the implant tightly joined with the surrounding bone, only a very slight histological inflammatory reaction occurred. Thus, CPC is a highly biocompatible material and seems to be safe for application in humans.

Animals↗
PubMed1997

Two metabotropic gamma-aminobutyric acid receptors differentially modulate calcium currents in retinal ganglion cells.

Metabotropic gamma-aminobutyric acid (GABA) receptors were studied in amphibian retinal ganglion cells using whole cell current and voltage clamp techniques. The aim was to identify the types of receptor present and their mechanisms of action and modulation. Previous results indicated that ganglion cells possess two ionotropic GABA receptors: GABAAR and GABACR. This study demonstrates that they also possess two types of metabotropic GABAB receptor: one sensitive to baclofen and another to cis-aminocrotonic acid (CACA). The effects of these selective agonists were blocked by GDP-beta-S. Baclofen suppressed an omega-conotoxin-GVIA-sensitive barium current, and this action was reversed by prepulse facilitation, indicative of a direct G-protein pathway. The effect of baclofen was also partially occluded by agents that influence the protein kinase A (PKA) pathway. But the effect of PKA activation was unaffected by prepulse facilitation, indicating PKA acted through a parallel pathway. Calmodulin antagonists reduced the action of baclofen, whereas inhibitors of calmodulin phosphatase enhanced it. Antagonists of internal calcium release, such as heparin and ruthenium red; did not affect the baclofen response. Thus, the baclofen-sensitive receptor may respond to influx of calcium. The CACA-sensitive GABA receptor reduced current through dihydropyridine-sensitive channels. Sodium nitroprusside and 8-bromo-cGMP enhanced the action of CACA, indicating that a nitric oxide system can up-regulate this receptor pathway. CACA-sensitive and baclofen-sensitive GABAB receptors reduced spike activity in ganglion cells. Overall, retinal ganglion cells possess four types of GABA receptor, two ionotropic and two metabotropic. Each has a unique electrogenic profile, providing a wide range of neural integration at the final stage of retinal information processing.

Action Potentials↗
PubMed1997

Expression of the brain creatine kinase gene is low in neuroblastoma cell lines.

The cytoplasmic creatine kinase (CKB) enzyme has a central role in the regeneration of ATP in the brain. We have shown previously that CKB mRNA levels in cultured primary rat brain astrocytes and oligodendrocytes are much higher than in primary neurons. It has been suggested that high CKB expression is essential for the energy-demanding functions of glial cells. Conversely, CKB may be repressed in most neuronal cells; however, CKB protein has previously been detected by immunohistochemistry in several distinct groups of neurons in the adult rodent brain. Presently, little is known of the factors responsible for the high CKB expression in glia and possible repression in neurons. In this report, we investigated if low CKB mRNA was characteristic of some established neuronal cell lines. CKB mRNA was found to be extremely low in mouse C1300 neuroblastomas NS20Y and N1E-115 but 10-fold higher in NG108-15, a hybrid cell composed of a C1300 neuroblastoma and a rat C6 glioma. Since we showed NG108-15 contained only rat CKB mRNA transcribed from the C6 glioma CKB gene, expression of CKB mRNA may be a manifestation of a glial property in NG108-15 cells. However, CKB mRNA expression in NG108-15 appeared not to be fully activated since it was still 5-fold lower than in (parental) C6 glioma and 10-fold lower than in cellular RNA from either total rat brain or cultured primary astrocytes. When neuronal differentiation was increased in NS20Y and N1E-115 by treating cells with prostaglandin E1 and theophylline, the extremely low CKB mRNA level was not significantly changed. In a comparative study, the CKB mRNA levels in NS20Y, N1E-115 and neuronal RT4-B8 and RT4-E5 cells (from the rat RT4 peripheral neurotumor) were at least 50-fold lower than that in C6 glioma and 100-fold lower than in cultured primary astrocytes. These cell lines may provide a system for the identification of factors involved in the possible repression of CKB in many neuronal cells.

Animals↗
PubMed1997

Expression of the brain creatine kinase gene in rat RT4 peripheral neurotumor cell lines and its modulation by cell confluence.

Creatine kinases (CK) catalyze the reversible transfer of a high energy phosphate group between creatine phosphate and ADP to regenerate ATP in cell types where the requirements for ATP are extensive and/or sudden. Previously, we have shown in primary rat brain cell cultures that brain CK (CKB) mRNA levels are highest in astrocytes and oligodendrocytes and much lower in neuronal cells. However, little is known of the factors which regulate CKB expression in the central nervous system and peripheral nervous system. To begin to investigate these factors, we asked in this report (1) if this pattern of CKB expression was also characteristic of some established glial and neuronal cell lines derived from the PNS; (2) whether CKB expression could be rapidly modulated by culture conditions, and (3) if CKB is expressed in cells with characteristics of glial cell progenitors. In subconfluent cells, CKB mRNA and enzyme activity were found to be high in both the rat RT4 peripheral neurotumor stem cell RT4-AC36A and its glial cell derivative RT4-D6. Conversely, CKB mRNA and activity were 5- and 8-fold lower, respectively, in the neuronal derivative RT4-E5 and, more dramatically, CKB was undetectable in neuronal RT4-B8 cells. Maintaining RT4-D6 glial cells at confluence rapidly increased CKB enzyme activity by 7-fold, such that D6 cells contained about 25% of the CKB level in lysates prepared from either whole adult rat brain or primary cultures of rat brain astrocytes. The levels of CKB mRNA and immunoreactive protein were also correspondingly increased in confluent D6 cells. These confluence-mediated increases in CKB appeared to be due to cell-cell contact and not the depletion of serum growth factors or an increase in intracellular cAMP. This study indicates that CKB expression is highest in cells displaying glial properties and can be rapidly modulated by appropriate culture conditions. The results are discussed in relation to the factors which may regulate CKB expression in vivo.

Animals↗
PubMed1997

Ectopic eye development in Drosophila induced by directed dachshund expression.

The dachshund gene encodes a nuclear protein that is required for normal eye development in Drosophila. In the absence of dachshund function, flies develop with severely reduced or no eyes. We show that targeted expression of dachshund is sufficient to direct ectopic retinal development in a variety of tissues, including the adult head, thorax and legs. This result is similar to that observed with the highly conserved Drosophila gene eyeless, which can induce ectopic eye formation on all major appendages. Here, we show that dachshund and eyeless induce the expression of each other and that dachshund is required for ectopic retinal development driven by eyeless misexpression. These results suggest that the control of eye development requires the complex interaction of multiple genes, even at the very highest regulatory levels.

Animals↗
PubMed1997

Defective endogenous nitric oxide-mediated modulation of cellular respiration in canine skeletal muscle after the development of heart failure.

BACKGROUND: It is well documented that nitric oxide (NO) suppresses the function of a number of mitochondrial enzymes. Our recent studies found that endogenous NO may play an important role in the modulation of tissue oxygen (O2) consumption and cellular respiration both in vitro and in vivo. METHODS: Tissue O2 consumption was measured by a Clark-type O2 electrode at 37 degrees C in freshly isolated skeletal muscle segments from the accessory head of the triceps brachii (90% type I muscle fiber) and extensor carpi radialis (86% type II muscle fiber) from normal dogs and dogs with tachycardia-induced heart failure. RESULTS: S-nitroso-N-acetylpenicillamine (SNAP), carbachol, and bradykinin at doses of 10(-7) to 10(-4) mol/L concentration significantly suppressed tissue O2 consumption both in the absence and presence of 2,4-dinitrophenol (1 mmol/L), a mitochondrial uncoupler. These effects were not significantly different in the accessory head of the triceps brachii (90% type I muscle fiber) and extensor carpi radialis (86% type II muscle fiber). The effects of carbachol and bradykinin but not SNAP were attenuated by NG-nitro-L-arginine (10(-4) mol/L), indicating inhibition of the formation of endogenous NO. The inhibitory effect on tissue O2 consumption in response to carbachol and bradykinin became significantly smaller in skeletal muscle from dogs with pacing-induced heart failure, but the effects of SNAP were unchanged. CONCLUSIONS: Endogenous NO released from microvascular endothelium may play an important physiologic role in the modulation of cellular respiration in skeletal muscle, and the loss of this regulatory function may contribute to peripheral metabolic disorders and poor exercise tolerance during heart failure.

2,4-Dinitrophenol↗