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Biomedical subjects

X B Wang

Publications and source records attributed to X B Wang.

At least 19 recordsLinked to original sources

Non-uniform spatial distributions of both the magnitude and phase of AC electric fields determine dielectrophoretic forces.

It is well known that the conventional dielectrophoretic force acting on a polarised particle in a non-uniform AC electric field is proportional to the in-phase component of the induced dipole moment and the non-uniformity of the field strength. In contrast, the travelling-wave-dielectrophoretic force that acts on a particle subjected to a travelling electric field is proportional to the out-of-phase component of the induced dipole moment. We derive a theory that unifies the description and interpretation of conventional dielectrophoretic and travelling-wave-dielectrophoretic forces. We show that a particle in a non-uniform AC electric field experiences a dielectrophoretic force due to spatial non-uniformities of the magnitude and the phase of the field interacting, respectively, with the in-phase and out-of-phase components of the induced dipole moment. The theory is used to explain the translational effects observed for particles in the presence of standing, travelling and rotating fields in several experimental electrode configurations. The good agreement found between the experimental observations and the theoretical predictions validate the theory.

Cell Movement

Separation of human breast cancer cells from blood by differential dielectric affinity.

Electrorotation measurements were used to demonstrate that the dielectric properties of the metastatic human breast cancer cell line MDA231 were significantly different from those of erythrocytes and T lymphocytes. These dielectric differences were exploited to separate the cancer cells from normal blood cells by appropriately balancing the hydrodynamic and dielectrophoretic forces acting on the cells within a dielectric affinity column containing a microelectrode array. The operational criteria for successful particle separation in such a column are analyzed and our findings indicate that the dielectric affinity technique may prove useful in a wide variety of cell separation and characterization applications.

Blood

Decrease in CRE binding activity by chronic morphine administration in mouse brain.

Recent studies have suggested that opiate addiction is associated with transcriptional changes. We developed a novel method, in situ DNA-protein binding (ISDB), for investigating the distribution and changes of DNA binding activity of transcription factors in the brain. Using this method, we found that cAMP response element (CRE) binding activity was decreased by chronic morphine treatment in specific regions including the amygdala complex, thalamus, cerebral cortex and hypothalamus in mouse brain. This effect persisted for at least 14 days after the cessation of morphine. These data suggest that chronic morphine treatment elicits a long-term change in cAMP-mediated gene expression in the brain.

Animals

Changes in Friend murine erythroleukaemia cell membranes during induced differentiation determined by electrorotation.

We used electrorotation measurements to investigate alterations in the plasma membranes of DS19 murine erythroleukaemia cells that accompanied erythropoietic differentiation induced by hexamethylene bisacetamide (HMBA). Following 3 days of HMBA treatment, the mean cell membrane specific capacitance determined from electrorotation spectra of individual, viable cells at physiological tonicity (300 mosmol/kg) fell from 1.74 to 1.53 microF/cm2, in agreement with trends observed earlier by dielectrophoretic measurements on bulk cell populations. Scanning and transmission electron microscopy revealed that the relatively high values found for cell membrane capacitance (> 1 microF/cm2) reflected the large area of plasma membrane associated with complex surface morphology including numerous microvilli. Furthermore, it demonstrated that the fall in membrane capacitance during HMBA treatment correlated with a reduction in the density of these complex surface features. Differences in the mechanical characteristics of the cell membranes of untreated and treated cells were then examined by exposing cells to osmotic stress. The intricacy of membrane morphology intensified with increasing osmolality of the suspending medium and this was reflected in higher specific capacitance values. When the osmolality was increased from 210 to 450 mosmol/kg, the mean membrane capacitance of untreated DS19 cells changed from 1.58 to 2.05 microF/cm2 while that for HMBA-treated cells changed from 1.47 to 1.72 microF/cm2, a significantly smaller response. This demonstrated that cells exposed to 72 h of differentiation treatment had an enhanced mechanical resilience as compared with their untreated counterparts, evidencing the early stages of the development of the membrane skeleton which becomes fully developed in mature erythrocytes. Our findings demonstrate the value of electrorotation measurements as a method for the non-invasive characterisation of viable leukaemic cells and their responses to stimuli and show that the membrane capacitance values so derived reflect membrane morphology.

Acetamides

Modulation by chronic morphine administration of single-stranded cAMP response element (ssCRE) binding proteins in the mouse cerebellum.

The development of opiate tolerance and dependence are thought to be associated with gene expression. Our previous studies have shown that the binding activity of nuclear factors to a single-stranded oligo-DNA containing cAMP response element (ssCRE) is altered by long term treatment with morphine in cultured neuronal cells. In the present experiments, the effects of acute and chronic treatments with morphine on the binding of nuclear proteins to single- and double-stranded oligo-DNAs of the cAMP response element were studied in the mouse brains by using gel shift assay. The activity of single-stranded CRE binding proteins (ssCRE-BP) was decreased by chronic morphine treatment to about 40% of control in the cerebellum. The effect of chronic morphine treatment on the binding activity persisted for at least 2 weeks after morphine withdrawal. The activity of double-stranded CRE binding proteins was also detected in the cerebellum, but it was insensitive to the morphine treatment. The activity of ssCRE-BP was also decreased by acute morphine treatment in 5 h, but it returned to control level in 24 h. These data suggest that the change of ssCRE-BP can be involved in the development of tolerance and dependence.

Animals

[Involvement of gene expression in drug tolerance and dependence].

The development of drug tolerance and dependence are thought to be associated with gene expression. Our studies showed that the binding activity of nuclear factors to several DNA sequences is altered by long-term treatment with methamphetamine, cocaine and morphine: 1) the binding activity of AP-1 increased markedly in the mouse brain after administration of methamphetamine and cocaine, 2) CRE-binding activity was decreased by chronic morphine treatment in the amygdala complex, cerebral cortex and hypothalamus of the mouse brain, and 3) the binding activity of single-stranded CRE binding proteins was decreased by chronic morphine treatment in the mouse cerebellum. These data suggest that the changes of DNA binding proteins can be involved in the development of drug tolerance and dependence.

Amphetamine

In situ DNA-protein binding: a novel method for detecting DNA-binding activity of transcription factor in brain.

A novel method, in situ DNA-protein binding (in situ DPB), was developed to detect the distribution and DNA-binding activity of AP-1 and Sp1 binding proteins in situ. The regional distribution of AP-1 binding protein in mouse brain was different from that of Sp1. Antibody against the DNA-binding domain of Jun protein markedly reduced the AP-1 but not the Sp1 binding activity. The binding activity of AP-1 probe increased markedly in the brain after administration of methamphetamine. These results suggest that the in situ DPB is convenient and sensitive for detecting the distribution and the DNA-binding activity of transcription factors in situ.

Animals

Medium hyperosmolarity inhibits prolactin secretion induced by depolarizing K+ in GH4C1 cells by blocking Ca2+ influx.

Medium hyperosmolarity between 300 (normal medium osmolarity) and 600 mOsm inhibited in a concentration-correlated fashion (r greater than 0.97, p less than 0.001) the rise in intracellular Ca2+ concentration ([Ca2+]i) and prolactin (PRL) secretion induced in GH4C1 cells by depolarizing 30 mM K+. [Ca2+]i concentration and PRL secretion were tightly related between 300 and 600 mOsm (r = 0.976, p less than 0.001); 50% inhibition of both occurred at 450 mOsm. Medium hyperosmolarity slowed the rate of Ca2+ influx. At 600 mOsm the rise in both [Ca2+]i and PRL secretion was abolished but PRL secretion induced by 1 microM phorbol 12-myristate 13-acetate was not significantly reduced. Our data suggest that inhibition of Ca2+ influx may be the primary mechanism by which extracellular hyperosmolarity inhibits PRL secretion induced by high medium K+ in GH4C1 cells. Depression of the Ca2+ intracellular transduction system may play a pathophysiological role in vivo in conditions such as dehydration and hypertonic coma.

Animals

Differences in the AC electrodynamics of viable and non-viable yeast cells determined through combined dielectrophoresis and electrorotation studies.

An electrode system is described for the near-simultaneous application and measurement of translational, levitational and rotational forces induced by AC electric fields, and this has been used to investigate the differences in the AC electrodynamics of viable and non-viable yeast cells. A new approach to the theoretical modelling of the experimental data has enabled these differences to be quantified in terms of changes in the conductivity of the cytoplasmic membrane and cell interior. The results are considered to have potentially important biomedical and biotechnological applications.

Cell Survival

Different natures of supersensitivity of adenylate cyclase stimulated by calcitonin gene-related peptide and isoproterenol in rat diaphragm after denervation and reserpine treatment.

In skeletal muscles, calcitonin gene-related peptide (CGRP) released from motor nerve terminals and humoral catecholamines stimulate adenylate cyclase (AC) and enhance muscle contraction. The effects of denervation and treatment with reserpine on twitch contraction and the AC system in rat diaphragm were investigated. The basal levels of twitch contraction and AC activity of the diaphragm of rats were both increased 2 weeks after phrenic nerve denervation but were not altered by treatment with reserpine. Reserpine treatment provoked supersensitivity of AC to isoproterenol, without affecting the response to CGRP. On the other hand, denervation decreased the activation of AC and enhancement of twitch contraction by CGRP, without affecting the responses to isoproterenol. These data suggest that denervation causes up-regulation of AC as a result of loss of CGRP release from nerve terminal and that depletion of catecholamines by reserpine treatment supersensitizes the responses at the beta-adrenoceptor level. Thus, nervous and humoral factors regulate the AC system in striated muscle by different mechanisms.

Adenylyl Cyclases

Different pathways for Ca2+ influx and intracellular release of Ca2+ mediated by muscarinic receptors in ileal longitudinal smooth muscle.

Muscarinic receptor-mediated elevations in intracellular Ca2+ concentration ([Ca2+]i) in the longitudinal smooth muscle of guinea pig ileum were studied by the use of fura-2 fluorescence. Dose-response analysis indicated a difference in the potencies of carbachol (CCh) to increase [Ca2+]i in the presence and absence of extracellular Ca2+. For the increase in [Ca2+]i due to Ca2+ release from intracellular stores in the absence of extracellular Ca2+, the ED50 value of CCh was 3 x 10(-5) M. On the other hand, in the presence of Ca2+, the ED50 value was 2.5 x 10(-7) M, indicating that a low concentration of CCh (less than 10(-7) M) caused influx of extracellular Ca2+ without Ca2+ release. Oxotremorine and pilocarpine induced Ca2+ influx, but were less potent inducers of Ca2+ release. CCh also stimulated the formation of inositol trisphosphates (IP3) with an ED50 value of (4.5 x 10(-5) M), which was similar to that for Ca2+ release from intracellular stores. Treatment of the smooth muscle with neomycin (1 mM), a phospholipase C inhibitor, abolished both CCh-induced IP3 formation and Ca2+ release from intracellular stores, but did not affect CCh-induced Ca2+ influx. These results suggest that the pathway for muscarinic stimulation of Ca2+ influx through plasma membranes is different from that for Ca2+ release from intracellular stores, which seems to be coupled with IP3 formation.

Animals

Clinical evaluation of 99Tcm-MIBI SPECT in the assessment of coronary artery disease.

To evaluate the clinical efficacy of 99Tcm-MIBI myocardial tomography for detecting coronary artery disease (CAD) 115 consecutive patients who underwent both 99Tcm-MIBI single photon emission computed tomography (SPECT) and coronary arteriography were studied. Thirty-three patients without and 82 with significant coronary artery disease were documented by coronary arteriography. The overall sensitivity and specificity of 99Tcm-MIBI SPECT for detecting coronary artery disease were 96 and 87.9%, respectively. The sensitivity for identifying patients with CAD without myocardial infarction was 88%. The sensitivity of 99Tcm-MIBI SPECT for detecting individual coronary artery lesions was 86% for left anterior descending artery (LAD), 69% for left circumflex artery (Lcx) and 86% for right coronary artery (RCA), lesions respectively. In conclusion, 99Tcm-MIBI SPECT provides a reliable method for detecting CAD.

Adult

Race, family income, and low birth weight.

The relations among race, family income, and low birth weight were examined using information obtained from the National Longitudinal Survey of Youth, which conducted yearly interviews with a nationally representative sample of young women identified in the late 1970s. Data were available for these women and their offspring from 1979 through 1988. Maternal education, maternal age, age/parity risk, marital status, and smoking during pregnancy served as covariates in cross-sectional and longitudinal analyses. The risk of low birth weight among births to black women and white women who were poor was at similarly high levels regardless of whether poverty was determined prior to study entrance or during the study period. Longitudinal analyses showed an exceptionally large increase in risk of low birth weight among children born to women whose prior pregnancy ended in a low-birth-weight infant. These two findings emphasize the importance of factors antecedent to the pregnancy in the genesis of low birth weight.

Adolescent

Evidence that potassium channels regulate prolactin secretion in GH4C1 cells by causing extracellular calcium influx.

Tetraethylammonium (TEA), a K+ channel blocker, induced prolactin (PRL) secretion in GH4C1 cells in a dose-dependent manner when applied at a concentration from 1-20 mM. During continuous exposure to TEA, a significant increase in PRL secretion occurred by 20 min and the response was sustained until the end of a 60-min exposure. Blocking Ca2+ influx by employing a Ca(2+)-depleted medium or the Ca2+ channel blocker, nifedipine, prevented induction of PRL secretion by 20 mM TEA. Preincubation of the cells for 10 min with 20 mM TEA did not inhibit PRL secretion induced by thyrotropin-releasing hormone (TRH), phorbol 12-myristate 13-acetate (TPA) or by cell swelling produced by 30% medium hyposmolarity, but significantly depressed that induced by depolarizing 30 mM K+. BaCl2, another K+ channel blocker, had the same effect on PRL secretion as TEA. The data suggest that blocking K+ channels may cause membrane depolarization, thereby inducing Ca2+ influx which is a potent stimulus for PRL secretion in GH4C1 cells.

Calcium

Dual effect of osmotic cell swelling on prolactin secretion by acutely dispersed adenohypophyseal cells.

Cell swelling induced by acute exposure to the permeant molecule urea or by medium hyposmolarity evoked a prompt PRL secretory burst from dispersed rat anterior pituitary cells. However, during continuous exposure greater than or equal to 10 min to these conditions inhibition of basal and TRH-induced PRL secretion occurred and there was an "off" burst of PRL secretion following return to basal conditions. Compared with continuous TRH stimulation which causes biphasic PRL secretion with a rapid high amplitude first phase secretory burst followed by a sustained low level second phase of secretion, cell swelling induced only "first phase" secretion. Removing Ca2+ from the medium or adding 50 microM verapamil markedly depressed the "off" secretory burst following return to basal conditions but had no effect on the initial high amplitude burst. Our data suggest that the effect of cell swelling on PRL secretion is complex and that there are at least two mechanisms for PRL secretion in normal anterior pituitary cells; these are differently affected by cell swelling and Ca2+ influx.

Animals

Dopamine inhibits cell swelling-induced prolactin secretion in MMQ cells by blocking Ca2+ influx.

To evaluate the role of Ca2+ influx on hormone secretion induced by cell swelling, we have utilized a prolactin (PRL)-secreting rat tumor cell line, MMQ, which has plasmalemma dopamine receptors. Medium hyposmolarity or osmotically equivalent isotonic urea caused prompt cell swelling and a rise in both [Ca2+]i and PRL secretion in a dose-dependent manner. Dopamine inhibited the induced increase in both [Ca2+]i and PRL secretion in a dose-dependent manner but the maximum inhibition was only 50%. This effect of dopamine was prevented by haloperidol. Depletion of medium Ca2+ or blocking Ca2+ influx with nifedipine completely abolished the osmotically induced rise in both [Ca2+]i and PRL secretion. These data indicate that Ca2+ influx through nifedipine-sensitive Ca2+ channels is an essential component of PRL secretion induced by osmotic cell swelling in MMQ cells and that a dopaminergic receptor-linked mechanism influences the opening of these channels.

Animals

Hormone secretion stimulated by ethanol-induced cell swelling in normal rat adenohypophysial cells.

Ethanol has been reported to affect endocrine functions, but its mechanism of action is unclear. To evaluate the hypothesis that cell swelling induced by ethanol permeation through the plasmalemma triggers hormone secretion, we studied the effect of ethanol on both hormone secretion and cell volume in acutely dispersed rat adenohypophysial cells under isotonic and hypertonic conditions. Isotonic ethanol caused a prompt cell swelling and an explosive secretory burst of prolactin and thyrotropin, which were proportional to the concentration of ethanol between 10 and 120 mM. The lowest effective dose of isotonic ethanol was 10 mM, which is below the plasma levels of legal intoxication (16 mM). Removal of medium Ca2+ enhanced the isotonic ethanol-induced increases in both cell volume and secretion. Hypertonic ethanol was ineffective in these effects. These data indicate that, in normal rat adenohypophysial cells, cell swelling caused by the rapid passage of ethanol through the plasmalemma is a potent mechanism for stimulating hormone secretion and this induced secretion is negatively modulated by extracellular Ca2+.

Animals

The contrasting role of calcium influx in secretion induced by cell swelling can differentiate normal and tumor-derived rat pituitary cells.

We have evaluated whether cell swelling may be a generally useful technique to differentiate normal and neoplastic pituitary cells, making the comparison between normal lactotrophs and thyrotrophs and tumor-derived GH4C1 and MMQ cells. With 1.5 mM medium Ca2+, cell swelling induced by osmotically equivalent stimuli, 27% medium hyposmolarity or 80 mM isotonic urea, caused a prompt increase in both intracellular Ca2+ and hormone secretion by all cell types. Depletion of medium Ca2+ abolished the cell swelling-induced increase in intracellular Ca2+ in all cell types and hormone secretion in the tumor-derived cells. However, it enhanced hormone secretion in normal cells. The critical role of Ca2+ influx in osmotically induced secretion in neoplastic, but not normal, pituitary cells may reflect some fundamental alteration in the intracellular transduction system in tumor cells.

Animals