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W X Shi

Publications and source records attributed to W X Shi.

35 records · Page 2Linked to original sources

Presence of antibodies to heat stress proteins and its possible significance in workers exposed to high temperature and carbon monoxide.

Antibodies to the ubiquitous group of stress proteins known as heat shock proteins (Hsps) have been found to be associated with a number of diseases in humans. Hsps are known to be induced by certain xenobiotics, some of which are common in the working environment. The biological significance of the presence of such autoantibodies is presently unclear. In the present study, we used immunoblotting to investigate the presence of antibodies against the different stress proteins, Hsp27, Hsp60, Hsp71, Hsc (heat shock cognate) 73 and Hsp89 alpha and beta in groups of workers exposed to high temperature or carbon monoxide. These data were related to a detailed clinical evaluation and to various laboratory measurements including electrocardiogram (ECG), B echogram, white blood cell counts and typing, the activity of alanine aminotransferase (ALT), acid phosphatase (ACP) and alkaline phosphatase (ALP) and lymphocyte DNA damage. Antibodies to Hsp27 and Hsp71 were found more frequently in the high temperature and carbon monoxide-exposed groups than in controls (P < 0.05). The carbon monoxide-exposed group showed the highest incidence of anti-Hsp antibodies. Anti-Hsp60 antibodies were only detected in workers exposed to high temperature or carbon monoxide. The percentage of workers with abnormal ECG, B echogram changes and displaying hepatitis B antigen (HBsAg) was higher in the carbon monoxide group than in the control group (P < 0.05). There was a significant increase in the activity of ALT in the high temperature and carbon monoxide groups and in the activities of ACP and ALP in the carbon monoxide group (P < 0.05). The extent of DNA damage measured in lymphocytes was higher in workers from the high temperature and carbon monoxide-exposed groups. We suggest that the increased frequency of antibodies to Hsps is the result of these damages of the release of denatured Hsps and of a decrease in the phagocytic ability of macrophages in these workers. The data gathered in the present study show a statistical relation between the occurrence of antibodies against Hsps and the frequency of health problems in workers and suggest a potential role for the antibodies as useful biomarkers to assess whether workers are experiencing environmental stress.

Antibodies↗

Nonlinear sequence-dependent structure of nigral dopamine neuron interspike interval firing patterns.

Firing patterns of 15 dopamine neurons in the rat substantia nigra were studied. These cells alternated between two firing modes, single-spike and bursting, which interwove to produce irregular, aperiodic interspike interval (ISI) patterns. When examined by linear autocorrelation analysis, these patterns appeared to reflect a primarily stochastic or random process. However, dynamical analysis revealed that the sequential behavior of a majority of these cells expressed "higher-dimensional" nonlinear deterministic structure. Dimensionality refers to the number of degrees of freedom or complexity of a time series. Bursting was statistically associated with some aspects of nonlinear ISI sequence dependence. Controlling for the effects of nonstationarity substantially increased overall predictability of ISI sequences. We hypothesize that the nonlinear deterministic structure of ISI firing patterns reflects the neuron's response to coordinated synaptic inputs emerging from neural circuit interactions.

Action Potentials↗

Ritanserin, a 5-HT2A/2C antagonist, reverses direct dopamine agonist-induced inhibition of midbrain dopamine neurons.

Ritanserin (RIT), widely-used as a selective 5-HT2A/2C receptor antagonist, has been reported to produce significant therapeutic effects on the negative symptoms of schizophrenia and to improve extrapyramidal side effects induced by neuroleptics. Because midbrain dopamine (DA) systems are believed to be the major site of action for many antipsychotic drugs, the effect of RIT on substantia nigra DA neurons was examined in chloral hydrate-anesthetized rats using single unit recording techniques. Systemic injection of RIT (0.1-6.4 mg/kg, i.v.) had no consistent effect on basal firing rate but significantly reversed the inhibition induced by both direct and indirect DA agonists. However, our data suggest that this effect of RIT is largely mediated by a mechanism independent of 5-HT. Thus the 5-HT2A/2C agonist 1(2,5 dimethyoxy-4-iodophenyl)-2-aminopropane showed no effect on either basal firing rate or the inhibition induced by the direct DA agonist quinpirole. Neither the selective 5-HT2A antagonist MDL 100907 nor depletion of endogenous 5-HT using p-chlorophenylalanine mimicked the effect of RIT (i.e., attenuated quinpirole-induced inhibition). Furthermore, the effect of RIT persisted in animals pretreated with p-chlorophenylalanine. Because RIT is known to bind D2-like receptors and because the inhibition of DA neurons induced by low doses of a direct DA agonist is believed to be mediated by DA autoreceptors, these results suggest that RIT may act on DA autoreceptors directly as a DA antagonist. Since similar doses of RIT were reported to have no significant effect on postsynaptic D2 receptors in the striatum, it is possible that RIT at the doses used may selectively block DA autoreceptors.

Amphetamines↗

A mannose-specific lectin from Vicia villosa seeds.

A lectin specific to mannose has been purified from Vicia villosa seed by (NH4)2SO4 fractionation, GalNAc-Sepharose and Man-Sepharose affinity chromatography. It was defined as VVLM, which showed a single band on an acidic-PAGE stained with Coomassie brilliant blue. The molecular weight of VVLM was 50 kDa as determined by gel filtration on Biogel P-100 column. The VVLM molecule consists of 2 distinct subunits with apparent molecular weight of 30 kDa and 22 kDa determined by SDS-PAGE. VVLM has at least four isolectins with similar haemagglutinating activity. Its extinction coefficient is calculated as A1(1cm) = 16.4 at 280 nm. Sugars could not be detected by phenol-sulfuric acid method. The circular dichroism analysis at far UV indicated that VVLM was a beta-sheet-rich protein, and gave no alpha-helix, 69% beta-sheet, 14% beta-turn by Provencher and Glockner method. The lectin was inhibited by alpha-methyl-D-mannose at 12.5 mM and glucose or GlcNAc at 50 mM. The carbohydrate binding specificity of VVLM was investigated by using affinity chromatography on a VVLM-Sepharose column. Among various Asn-linked oligosaccharides, core structure Man alpha 1-->3(Man alpha 1-->6)Man beta 1-->4GlcNAc beta 1-->4GlcNAcOT were found to have high affinity for VVLM-Sepharose. The antisera of VVLM did not produce precipitin line with VVLG in agar double diffusion plate indicating no serological relationship between VVLM and VVLG. However VVLM showed similar immunodeterminants of some other lectins of mannose specificity such as Con A, PSL, LCA and VFL.

Amino Acids↗

GABA synapses formed in vitro by local axon collaterals of nucleus accumbens neurons.

GABAergic medium-spiny neuron axons not only form the principal projections of the nucleus accumbens (nAcc) but also branch locally in a dense network overlapping their own dendrites, suggesting that their recurrent synapses mediate the major information processing functions of the nAcc. We used postnatal nAcc cultures to study these synapses individually. In culture, as in the intact nAcc, medium-spiny neurons account for over 95% of the cells and are GABAergic. Strikingly, these neurons showed a spike afterhyperpolarization (AHP) that was largely blocked by the GABAA antagonist bicuculline. The bicuculline-sensitive AHP occurred without or with latency, and met criteria for monosynapticity; consistent with this, dye fills showed the presence of recurrent axons and a low incidence of dye coupling. Blockade of Ca2+ influx eliminated this autaptic PSP, while TTX almost completely eliminated it, indicating that it is due to exocytic GABA release principally at axodendritic contacts. While blocking GABAB receptors had no direct effect on the autaptic PSP, activating these receptors with baclofen produced presynaptic inhibition, as well as directly mediated hyperpolarization; together, these actions increased the signal-to-noise ratio in the cellular response to synaptic inputs. Bicuculline also increased the signal-to-noise ratio; in addition, it induced burst firing and depolarization inactivation. In contrast, the indirect GABA agonist flurazepam and the GABA uptake blocker nipecotic acid each enhanced autaptic PSPs. Since autapses formed in vitro appear to be functionally equivalent to synapses between neighboring medium-spiny neurons that receive similar inputs, these results bear on the function of intrinsic GABA synapses in the intact nAcc. Thus, intrinsic GABA synapses are likely to regulate the signal-to-noise ratio in nAcc information processing and may be important targets for the modulatory actions of endogenous neurotransmitters and drugs.

Animals↗

Conformation and activity of Phaseolus coccineus var. rubronanus lectin.

The conformation of native and denatured Phaseolus coccineus var. rubronanus lectin was studied by circular dichroism (CD) and correlated to the hemagglutinating activity. The far-UV CD spectrum at 25 degrees C showed a broad, negative band around 223 nm and a positive one at 196 nm. CD data analysis of the lectin indicated a beta-sheet-rich protein. At high temperatures, the spectrum was blue-shifted with increasing magnitude; these changes correlated well with the loss of the activity. The conformation of lectin between pH 2 and 10 remained essentially unchanged. At pH 13 the CD spectrum resembled that of unordered form with a negative band near 200 nm and the activity was completely lost. The denatured lectin in 6 M guanidine hydrochloride would be renatured upon diluting the denaturant to 0.75 M; the changes in CD spectrum again correlated well with the loss of the activity. The effect of sodium dodecyl sulfate on the lectin was drastic; it sharply increased the alpha-helix at the expense of the beta-sheet and reduced the activity; the changes reached a plateau above 20 mM surfactant.

Circular Dichroism↗

Conformation and activity of mannose- and N-acetylgalactosamine-specific lectins from Vicia villosa seeds.

The conformation of two Vicia villosa lectins specific for mannose and N-acetylgalactosamine, respectively, was studied by circular dichroism. Both showed a broad negative CD band around 220 nm and a positive one above 190 nm. CD data analysis indicated that they were rich in beta-sheet. However, they differed in conformational stability against extreme pH, at elevated temperature, and in guanidine hydrochloride and sodium dodecyl sulfate solutions. The unusual feature was that the conformation of N-acetylgalactosamine-specific lectin was virtually unaltered in 6 M guanidine hydrochloride and 7.5 mM surfactant.

Acetylgalactosamine↗

Actions of neurotensin: a review of the electrophysiological studies.

Three effects of NT were observed on midbrain DA cells. The modulatory effect of NT, that is, the attenuation of DA-induced inhibition, has been most extensively examined. Studies indicate that this effect of NT was not simply due to a nonspecific excitation. NT selectively attenuated DA-induced inhibition without affecting either GABA-induced inhibition or glutamate-induced excitation of the same cells, and the attenuation of DA-induced inhibition could be observed at the doses at which the basal activity of DA cells was not changed by NT. The attenuation of DA-induced inhibition by NT is also unlikely to result from the formation of a DA-NT complex, since neuromedin N, which competes with NT for the same receptor but does not bind to DA, mimicked the effects, and neurotensin(1-11), which forms a complex with DA but is inactive in competing for NT receptors, did not. The similarities between the effects of NT and those of 8-bromo-cAMP and forskolin suggest that intracellular cAMP and protein kinase A may be involved. This suggestion was supported by the findings that IBMX (an inhibitor of phosphodiesterases) potentiated the effect of NT; and SQ22536 (an inhibitor of adenylate cyclase) and H8 (an inhibitor of protein kinase A) antagonized it. Phorbal-12,13-dibutyrate (an activator of protein kinase C) did not mimic the effect of neurotensin, and H7 (an inhibitor of protein kinase C) did not reduce the effect, suggesting that protein kinase C is unlikely to be involved in the modulatory effect of neurotensin. Experiments in vitro indicated that the excitatory effect of NT on DA cells occurred at higher concentrations (> 10 nM) than those needed for producing the modulatory effect. Its persistence during DA receptor blockade by sulpiride suggests that this effect was not entirely mediated by an attenuation of the inhibition induced by endogenously released DA. At even higher concentrations (> 100 nM), a sudden cessation of cell activity preceded by an increase in firing rate was observed. Whether this effect of NT was due to depolarization inactivation or a toxic effect of the peptide at high concentrations remains to be determined. In most other areas studied, the excitatory effect of NT was most commonly observed. In many areas, this excitatory effect was apparently a direct postsynaptic effect of NT. However, different mechanisms may be involved (see Table 1). For example, in some areas NT acted through a decrease in membrane conductance, while in others no change or an increase in the membrane conductance was observed.(ABSTRACT TRUNCATED AT 400 WORDS)

Neurotensin↗

Roles of intracellular cAMP and protein kinase A in the actions of dopamine and neurotensin on midbrain dopamine neurons.

The role of intracellular cAMP and protein kinase A in dopamine-induced inhibition of dopamine neurons and the attenuation of this inhibition by neurotensin were studied in rat midbrain slices. Spontaneous activity of dopamine cells was recorded extracellularly from both the ventral tegmental area and the substantia nigra. Perfusion of the slices with 8-bromo-cAMP and forskolin significantly attenuated dopamine-induced inhibition, but neither blocked the inhibition completely. Neither SQ22536, an inhibitor of adenylate cyclase, nor H8, an inhibitor of protein kinase A, mimicked the inhibitory effect of dopamine on dopamine neurons, although they potentiated dopamine's effect. These results indicate that dopamine-induced inhibition of dopamine neurons can be affected by intracellular cAMP levels, but is unlikely to be mediated solely by inhibition of adenylate cyclase. The similarities between the effects of neurotensin and those of 8-bromo-cAMP and forskolin suggest that intracellular cAMP may be involved in the actions of neurotensin. This suggestion is supported by our findings that isobutyl-methylxanthine (an inhibitor of phosphodiesterases) potentiated the effect of neurotensin and SQ22536 and H8 antagonized it. Phorbol-12,13-dibutyrate (an activator of protein kinase C) did not mimic the effect of neurotensin, and H7 (an inhibitor of protein kinase C) did not reduce the effect, suggesting that protein kinase C is unlikely to be involved in the modulatory effect of neurotensin.

Animals↗

Identified postnatal mesolimbic dopamine neurons in culture: morphology and electrophysiology.

To examine the intrinsic properties of postnatal mesolimbic dopamine (DA) neurons, we dissociated the ventral tegmental area (VTA) from postnatal rats, enriched for DA neurons by microdissection or gradient purification, and grew the cells in culture. In these cultures, up to 50% of neurons were dopaminergic. DA neurons resembled their in vivo counterparts in soma shapes, and in showing two levels of tyrosine hydroxylase (TH) expression, axodendritic differentiation, two sizes of synaptic vesicles, nest-like synaptic arrangements with non-DA cells, and synaptic specializations. Electrophysiologically, however, they could not be distinguished from non-DA cells, which could be consistent with heterogeneity in cell properties. To examine a functional subset of VTA DA neurons, we retrogradely labeled VTA neurons projecting to the nucleus accumbens. These mesoaccumbens neurons were 86% TH positive, 56% cholecystokinin positive, and 0% neurotensin positive; they also displayed the soma shapes characteristic of DA neurons more generally and two levels of TH expression. Like their in vivo counterparts, mesoaccumbens cells generally fired single broad spikes that were triggered by slow depolarizations and had robust spike afterhyperpolarizations, low- and high-threshold Ca2+ spikes, rapid accommodation of firing, time-dependent anomalous rectification, and hyperpolarizing autoreceptor responses. Strikingly, the expression of these active properties did not change with time in culture. Mesoaccumbens DA cells could be identified by a distinctive subset of properties that made up an electrophysiological signature; however, unlike their in vivo counterparts, they were less often spontaneously active and never fired in bursts. These results suggest that most DA cell properties are intrinsic to the cells, including a significant heterogeneity that is maintained in postnatal culture; their level and mode of activity, however, appear to require afferent input. Culturing identified postnatal VTA DA neurons now makes possible examination of the impact of their individual properties on synaptic function.

Animals↗

Neurotensin modulates autoreceptor mediated dopamine effects on midbrain dopamine cell activity.

Interactions of neurotensin (NT) with midbrain dopamine (DA) neurons were studied in rats using microiontophoretic techniques. Local ejection of NT significantly increased (greater than 30%) the firing rate of a few DA cells (12/106). In most cases, however, iontophoretic NT produced no significant change in spontaneous activity. On the other hand, in these same cells, NT significantly attenuated the inhibition induced by either DA or quinpirole, a specific D2 agonist. Inhibition induced by DA was not attenuated by either glutamate or cholecystokinin, although both of them increase the firing rate of DA cells. The effect of NT appears to be selective as NT attenuated DA-induced inhibition without a measurable effect on either GABA-induced inhibition or glutamate-induced excitation of the same DA cells. Combined, these results suggest that NT's effect on DA cell activity is primarily a neuromodulatory one. As both NT and D2 receptors in midbrain DA cell areas are primarily located on DA cells, the above results also suggest that the observed interaction between NT and DA occurred at the DA cell level.

Animals↗

Effects of neurotensin on midbrain dopamine neurons: are they mediated by formation of a neurotensin-dopamine complex?

The effects of neurotensin on midbrain dopamine neuron activity were studied in brain slices using single-unit recording techniques. At low concentrations (0.2-10 nM), neurotensin attenuated dopamine-induced inhibition without a significant effect on the basal firing rate. At higher concentrations (greater than 10 nM), however, it consistently caused an increase in cell activity. At even higher concentrations (greater than 100 nM), a sudden cessation of cell activity preceded by an increase in firing rate was observed. Whether this effect of neurotensin was due to depolarization inactivation or to a toxic effect of the peptide at high concentrations remains to be determined. To determine whether the effects of neurotensin were mediated by formation of a neurotensin-dopamine complex, several neurotensin analogues were studied. Neurotensin (8-13), which binds to both neurotensin receptors and dopamine, mimicked the effects of native neurotensin. Neuromedin N, which competes with neurotensin for the same receptor but does not bind to dopamine, also mimicked the effects. However, neurotensin (1-11), which forms a complex with dopamine but is inactive in competing for neurotensin receptors, was ineffective. In addition, the excitatory effect of neurotensin was not attenuated in the presence of dopamine receptor blockade by sulpiride. These results suggest that formation of a neurotensin-dopamine complex may not account for the action of neurotensin on dopamine cells. When combined with the fact that there is a high density of neurotensin receptors on dopamine cells, our results support the suggestion that the observed effects of neurotensin on dopamine neurons are most likely mediated by an activation of neurotensin receptors.

Animals↗

HLA-A, -B, -DR, -DQ antigens in black North Americans.

HLA antigens of black North Americans are not as well-characterized as those of other ethnic groups. We present results from HLA typing of black Americans using the 10th International Histocompatibility Workshop (IHW) sera. 1502 black Americans are typed for HLA-A, -B antigens. 128 black Americans are typed for HLA-DR, -DQ antigens. New specificities and splits established by the 10th IHW are identified in our data. Antigen frequency, gene frequency and haplotype frequency are computed for each phenotype observed. Haplotype A30--Bw42--DRw18--DQw4 and haplotype Aw36--Bw53 in strong linkage disequilibrium is characteristic of this black population. DR and DQ association patterns are similar to those of white North Americans. A major difference is the association of DRw8 with DQw7 in blacks versus an association of DRw8 with DQw4 in whites.

Black People↗

A simple and effective method for preventing the formation of salt bridges between barrels of a multibarrel microiontophoresis electrode.

Several kinds of multibarrel glass blanks are available commercially for making a multibarrel microiontophoresis electrode. Among them are the ones that consist of fused straight lengths of tubing available in three to nine barrels. Although they enjoy many advantages over the other kinds of assemblies, salt bridges form easily between the barrels of the electrodes made from these blanks. These bridges, when they form, render the multibarrel pipette unusable. Here, we describe a simple and effective way to prevent the formation of salt-bridge by coating the end of the barrels with paraffin. The paraffin coated electrode exhibits excellent recording properties which are equivalent to those made from more expensive blanks.

Animals↗

A small volume chamber for electrical recording from submerged brain slices and a pulse-free medium supply system using a peristalic pump.

A small volume chamber for recording from submerged brain slices is described. In the chamber, the slice is covered on all sides by a thin layer of moving medium. The chamber is simple and easy to construct. The small volume of the chamber allows fast exchange of the medium solution surrounding the slice and reduces the complications induced by long time exposure of slices to certain drugs (eg. desensitization). Along with this small volume chamber, a medium supply system is also described, which is composed of a peristaltic pump and a pulse dampener. This pulse-free medium supply system allows cells to be held for up to 8 h with solution changes as frequently as every 10-20 min.

Animals↗

Tetrahydroprotoberberine--a new chemical type of antagonist of dopamine receptors.

In rats lesioned by unilateral micro-injection of 6-OHDA into substantia nigra, the apomorphine-induced contralateral rotation and the amphetamine-induced ipsilateral rotation were antagonized by THB, l-THP and haloperidol. Scopolamine reversed the antagonistic effect of THB against amphetamine. Thus, THB and l-THP exhibited the DA-receptor antagonistic property which was similar to that of haloperidol. l-SPD (10 mg/kg), however, only antagonized the amphetamine-challenged rotational response, while it could not antagonize, but potentiate, the apomorphine-challenged rotational response. So, l-SPD might be a partial agonistic antagonist of DA-receptors, and its effect was more potent than that of THB and l-THP. l-SPD would be ascertained further in clinic trial. From these results and others, the authors suggest that THPB is a new chemical type of antagonist of brain DA-receptors.

Alkaloids↗