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Biomedical subjects

Y H Tan

Publications and source records attributed to Y H Tan.

At least 73 records · Page 4Linked to original sources
PubMed1992

Protein phosphatase inhibitors induce the sustained expression of the Egr-1 gene and the hyperphosphorylation of its gene product.

The immediate-early gene Egr-1 is strongly and rapidly induced in human and mouse Balb/c fibroblasts by okadaic acid and calyculin A, both specific inhibitors of protein serine/threonine phosphatases 1 and 2A. In contrast to the transient induction of the Egr-1 gene by serum or phorbol 12-myristate 13-acetate, these phosphatase inhibitors stimulated a sustained induction of the Egr-1 gene. The induction is shown to occur transcriptionally and is sustained post-transcriptionally. Okadaic acid-induced Egr-1 mRNA is significantly more stable than serum-induced Egr-1 mRNA. The half-life of serum-induced Egr-1 mRNA is estimated to be 12 min, compared with a half life of 2 h for okadaic acid-induced Egr-1 mRNA. Okadaic acid also induced the expression of the related immediate-early genes Egr-2 and Egr-3 albeit to a lesser extent than Egr-1. Treatment of cells with okadaic acid and calyculin A also induced the synthesis of Egr-1 protein. The Egr-1 protein is weakly or not phosphorylated in quiescent cells, but multiple species of the phosphorylated forms of the Egr-1 protein are detected in cells treated with either of the phosphatase inhibitors. Simultaneous treatment of cells with TPA and okadaic acid synergistically induced Egr-1 gene expression, and H7 strongly inhibits this induction. Taken together, the results indicate that the induction of Egr-1 gene transcription and the phosphorylation of the induced Egr-1 protein are under the control of protein kinase(s) and protein phosphatase(s). The phosphorylation and dephosphorylation of Egr-1 protein may play a role in controlling cell growth.

3T3 Cells↗
PubMed1992

Okadaic acid mimics multiple changes in early protein phosphorylation and gene expression induced by tumor necrosis factor or interleukin-1.

Okadaic acid, a phosphatase inhibitor from a marine organism, mimics tumor necrosis factor/interleukin-1 (TNF/IL-1) in inducing changes in early cellular protein phosphorylation. A total of approximately 116 proteins exhibit significant and concordant changes in phosphorylation or dephosphorylation within 15 min in human fibroblasts activated by either okadaic acid, TNF, or IL-1. The fidelity of this mimicry by okadaic acid extends to the phosphorylation of the 27 hsp complex, stathmin, eIF-4E, myosin light chain, nucleolin, epidermal growth factor receptor, and other cdc2-kinase substrates (c-abl, RB, and p53). The okadaic acid-induced pattern of protein phosphorylation is distinct from that observed in cells treated with phorbol 12-myristate 13-acetate or with ligands like epidermal growth factor, cyclic AMP agonists, bradykinin, or interferons. Like TNF, okadaic acid also induces the transcription of immediate early response genes like c-jun and Egr-1 as well as the interleukin-6 genes. The overall early effects of okadaic acid uniquely parallel those of TNF/IL-1 and not those of other cytokines or ligands. Regulation of protein phosphatase inhibition is discussed as a mechanism for TNF/IL-1 signal transduction.

Autoradiography↗
PubMed1992

Regulation of the Egr-1 gene by tumor necrosis factor and interferons in primary human fibroblasts.

Treatment of quiescent primary human fibroblasts with tumor necrosis factor (TNF) alpha, TNF-beta, interleukin-1, interferon (IFN) alpha, IFN beta, or IFN gamma induced Egr-1 mRNA. In primary human fibroblasts TNF-alpha and TNF-beta were mildly mitogenic and IFN alpha and IFN gamma were growth inhibitory. However, in HeLa cells TNF but not IFN induced the expression of Egr-1 mRNA, while both cytokines inhibited HeLa cell division. Kinetic measurements of Egr-1 gene expression showed that TNF-alpha, TNF-beta, and IFN gamma increased the cellular concentration of Egr-1 mRNA within 30 min. A maximum induction of Egr-1 mRNA was detected at approximately 60 min which dropped to basal level by 180 min. Induction was inhibited by H7 and staurosporine but not by HA1004, indicating the involvement of a functional protein kinase C. The Egr-1 message was translated and the cellular Egr-1 protein detected within 60 min of cytokine treatment. Despite similar Egr-1 mRNA induction, the amount of Egr-1 protein translated in IFN alpha- and IFN gamma-treated cells was lower than in those treated with TNF-alpha and TNF-beta, and highest in the EGF-treated primary human fibroblasts. Indeed, the level of Egr-1 protein translated in these cells correlated proportionally with both the phosphorylation of cap-binding protein (eukaryotic initiation factor) and the amount of cellular DNA synthesis in the variously treated fibroblasts. These results suggest that both growth stimulatory and inhibitory cytokines can regulate Egr-1 gene expression at the transcriptional and translational level. However, the combination of these regulatory controls may determine the cellular concentration of the Egr-1 gene product and hence, its effect on cell proliferation.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1992

Full-length cDNA sequence of dengue type 1 virus (Singapore strain S275/90).

The complete nucleotide sequence and the deduced amino acid sequence of the genome of dengue virus type 1 (Singapore strain S275/90) were determined from cDNA clones. The single-stranded, positive-sense RNA is 10,718 nucleotides in length and contains a single long open reading frame of 10,188 nucleotides encoding a polyprotein of 3396 amino acids. The genomic size and organization were found to be similar to that of other dengue virus serotypes. Both the nucleotide and deduced amino acid sequences were compared with the partial sequence of DEN1 (Nauru Island) and complete sequences of DNE2 (Jamaica), DEN3 (H87), and DEN4 (Dominica) virus genomes.

Amino Acid Sequence↗
PubMed1992

Protein phosphatases in cell signalling.

Protein phosphorylation is the most common post-translational modification and plays a role in all known pathways of signal transduction. Net phosphorylation is a result of the balance of activities of protein kinases and phosphatases. There is increasing evidence that the regulated removal of phosphate groups from proteins, catalyzed by protein phosphatases, is required for the downstream activation of other signalling proteins.

Animals↗
PubMed1992

Interleukin-6 regulates the cytotoxic effect of tumour necrosis factor on U937 cells.

In this study, we demonstrate that low but not high concentrations of interleukin-6 (IL-6) potentiate the cytotoxic effect of tumour necrosis factor-alpha (TNF-alpha) on U937 cells, in a dose-dependent manner. Killing of U937 cells by 100 U/ml of TNF-alpha, was maximally potentiated by 50 U/ml of IL-6. No potentiation of cell killing was observed when the concentration of IL-6 was increased to 4000 U/ml. At a concentration of 50 U/ml, IL-6 up-regulated TNF receptor expression but no change in TNF receptor number was observed when the concentration of IL-6 was increased to 4000 U/ml. Low concentrations of IL-6 can also induce sub-cytotoxic doses of TNF-alpha (0.1 and 0.33 U/ml) to kill U937 cells. Up-regulation of TNF receptors by IL-6 is dependent on de novo protein synthesis since receptor induction is abolished in the presence of cycloheximide. Taken together the data suggest that the potentiation of cell killing observed by a combination of these lymphokines is mediated in part by IL-6-induced changes in TNF receptor expression.

Cell Line↗
PubMed1992

Protective effect of cycloprotobuxine-A against cardiac arrhythmias induced by ouabain.

Cycloprotobuxine-A (CPB-A) 1-4 mg.kg-1 iv increased the dose of ouabain required to induce ventricular arrhythmias in guinea pigs. At the equitoxic doses (1/50 LD50), CPB-A was more potent than cyclovirobuxine-D and amiodarone. Pretreatment with reserpine (5 mg.kg-1 ip), vagotomy or pithing spinal cord did not prevent the action of CPB-A, which indicate that the protective effect of CPB-A may be due to its direct action on myocardium without the involvement of nervous system. In isolated guinea pig ventricular muscles, CPB-A 3 mumol.L-1 consistently decreased the amplitude of oscillatory afterpotentials (OAP) and blocked triggered activity elicited by ouabain. At 30 mumol.L-1, CPB-A abolished the appearance of OAP. It seems that one of the mechanisms for the anti-arrhythmic action of CPB-A was a decrease in the amplitude of OAP.

Alkaloids↗
PubMed1991

Histamine H3-receptors inhibit sympathetic neurotransmission in guinea pig myocardium.

The histamine H3 agonist, (R)-alpha-methylhistamine (alpha-MeHA, 10(-10) to 10(-5) M), caused a concentration-dependent inhibition of the sympathetic contractile response to electrical field stimulation of guinea pig isolated atria, but alpha-MeHA did not alter the basal tension or the contraction induced by exogenously applied norepinephrine. Blockade of H1 and H2 histamine receptors, and alpha- and beta-adrenoceptors failed to prevent the inhibitory effect of alpha-MeHA, whereas the specific H3 receptor antagonist, thioperamide, concentration dependently reversed the inhibitory effect of alpha-MeHA. At the concentration of 10(-7) M, which was effective for antagonizing the action of alpha-MeHA, thioperamide did not modify the sympathetic responses facilitated by the beta 2-adrenoceptor agonist, clenbuterol, or attenuated by the alpha 2-adrenoceptor agonist, clonidine. Our results suggest that H3 receptors exist on the cardiac sympathetic terminals, which may modulate adrenergic neurotransmission in guinea pig myocardium.

Animals↗
PubMed1991

Interleukin 1 and tumor necrosis factor activate common multiple protein kinases in human fibroblasts.

High resolution two-dimensional gel electrophoresis was used to analyze the signal transduction pathways of tumor necrosis factor (TNF-alpha) and interleukin 1 (IL-1 alpha and -beta) in human fibroblasts. Approximately 450 discrete radioactive spots were electrophoretically resolved from cytosolic extracts of cells prelabeled with 32P. At least 63 of these polypeptides exhibited significant and concordant phosphorylation or dephosphorylation in response to TNF or IL-1, despite the fact that different receptors are involved. Most of these changes concerned serine/threonine residues although enhanced tyrosine phosphorylation of several polypeptides was also observed. Phosphorylation patterns induced by a number of other agonists were compared with the patterns induced by IL-1 and TNF. These included activators of protein kinases C and A, bradykinin (a stimulator of inositol phospholipid hydrolysis), epidermal growth factor, heatshock, and mellitin (an activator of phospholipase A2). Although each of these agonists induced changes resulting in a distinct pattern of protein phosphorylation, none of these patterns had significant homology with that induced by IL-1 and TNF. Other assays were performed to verify the involvement of specific kinases. Collectively, these data indicate that IL-1 and TNF activate multiple protein kinases viz. a kinase(s) which activates microtubule-associated protein 2 (MAP-2) kinase, a kinase that phosphorylates the cap-binding protein, and a possibly novel serine/threonine protein kinase.

Autoradiography↗
PubMed1991

Small-vessel pathology and anastomosis following maxillofacial firearm wounds: an experimental study.

This experiment was designed to provide a biological basis for early reconstruction of maxillofacial defects caused by firearms using free vascularized flap transfer. The pathologic changes of small-vessel injury, injury mechanism, and the feasibility for early anastomosis of these vessels in the maxillofacial region following high-velocity missile wounds were studied. Dogs' faces were wounded by a steel sphere, weighing 0.7 g and 1.03 g, with an initial velocity of 1,300 m/s or 1,500 m/s. Angiography and high-speed x-ray photography were used to record the effect of injury to blood vessels and other tissues. The pathologic changes in the small blood vessels in the wounds were studied by light microscopy (LM) and electron microscopy (EM). Anastomosis of small vessels in the wound was performed at different times after injury. There were temporary cavity effects and small vascular injury in the wounds. The pathologic changes of the small vessels included microthrombus formation, endothelial loss, breaks in the internal elastic layer, and necrosis. Degeneration of cells was found to end 3 cm from the wound edge. Anastomosis of the vessels performed 3 days after injury provided good short-term patency. These results suggest that the operation for repairing facial defects should be performed 3 days after injury.

Animals↗
PubMed1991

Do second messengers play a role in interferon signal transduction?

The signalling pathway by which the binding of interferons (IFNs, alpha and beta) to their receptor elicits its biological activity, the activation of the transcription of a distinct set of genes called the IFN-stimulated genes (ISG), is far from clear. Debate continues about whether interferon-receptor interaction results directly in gene activation or if second messengers are involved. In this article, we will discuss the evidence that rapid and transient changes in lipid metabolism and the activation of specific isoforms of protein kinase C (PKC) are involved in the initial signalling of interferon activation.

Animals↗
PubMed1991

Effect of sodium chloride on transport of bacteria in a saturated aquifer material.

Determinations were made of the influence of NaCl concentration, cell density, and flow velocity on the transport of Pseudomonas sp. strain KL2 through columns of aquifer sand under saturated conditions. A pulse-type boundary condition was used. The experiments were conducted by using 0.3-m-long Plexiglas columns with an internal diameter of 0.05 m. When a 1-h pulse of a 0.01 M NaCl solution containing 10(8) cells per ml was added at a flow rate of 10(-4) m s-1, the bacterial density in the effluent never exceeded 2.2% of the density of cells added, and only 1.5% of the bacteria passed through the aquifer material. In contrast, when the bacteria were applied in distilled water, the relative cell density in the effluent approached 100%, and 60% of the bacteria were transported through the aquifer solids. Under these conditions, the breakthrough of Pseudomonas sp. strain KL2 was slower than chloride. When the flow rate was 2.0 x 10(-4) m s-1, the cell density in the effluent reached 7.3% of that added in 0.01 M NaCl solution, but only 3.9% of the bacteria were transported through the aquifer particles. On the other hand, the density in the effluent approached 100% of that added in deionized water, and 77% of the added bacteria were recovered. When the density of added cells was 10(9) cells per ml at a flow rate of 10(-4) m s-1, the densities in the effluent reached 70 and 100% of those added in salt solution and deionized water, respectively, and 44 and 57% of the bacteria were transported through the aquifer solids.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Movement↗
PubMed1990

Arachidonic acid regulates the binding of human interferon in human skin fibroblasts.

The induction of the antiviral state in human fibroblasts by human interferon is inhibited by arachidonic acid, its analogues 5,8,11,14-eicosatetraynoic and 5,8,11-eicosatriynoic acids, as well as by sodium arachidonate. The fatty acids myristic or oleic acid and sodium palmitate do not inhibit the antiviral action of interferon. Experiments were conducted to investigate the mechanism by which arachidonic acid could inhibit the action of interferon. No correlation between cellular lipoxygenase activities and the inhibition of antiviral action of interferon was observed in the fatty acid treated cells. Likewise, the cyclooxygenase inhibitors indomethacin and oxyphenylbutazone do not inhibit the interferon-induced antiviral state. Taken together, the inhibition of interferon action by arachidonates is unlikely to be mediated by cyclooxygenase or lipoxygenase-generated intermediates, even though arachidonates are known to affect the activity of these enzymes in vitro. Measurement of interferon receptors in the fatty acid treated cells showed that arachidonic acid, sodium arachidonate and its analogues decreased the number of human type I interferon receptors available for binding, and inhibited the transcription of the interferon-induced 6-16 gene and the induction of cellular (2'-5')-oligoadenylate synthetase, suggesting the mechanism of inhibition is mediated at the level of the interferon receptor. The significance of the finding that arachidonic acid, a common fatty acid of cells and serum, can affect the antiviral action of interferon is discussed.

2',5'-Oligoadenylate Synthetase↗
PubMed1990

[Effects of ranitidine and cimetidine on automaticity in isolated myocardium of guinea pig].

At 0.1-30 mumols/L, neither ranitidine (Ran) nor cimetidine (Cim) inhibited the rate of spontaneous contraction in isolated right atria of guinea pig. When the concentration was higher than 100 mumols/L, both Ran and Cim exhibited weak negative chronotropic effects with IC50 of 109 +/- 3 and 436.5 +/- 1.9 mumols/L, respectively. However, the positive chronotropic effects of histamine (H) were markedly antagonized by Ran and Cim with IC50 of 0.40 +/- 0.29 and 1.8 +/- 0.6 mumols/L respectively. Ran (0.1 mumols/L) and Cim (1 mumols/L) competitively antagonized the concentration-dependent response of positive chronotropic effect mediated by H and had no influence on that induced by isoproterenol in right atria of guinea pig. Ran (10 mumols/L) and Cim (50 mumols/L) prevented the abnormal automaticity elicited by H and H (0.12 +/- 0.09 mumols/L) + Oua (0.04 +/- 0.02 mumols/L) which acted synergically. The abnormal automaticity induced by Oua (1.5 +/- 2.3 mumols/L) was antagonized by Ran (10 mumols/L) and not by Cim (50 mumols/L). Our results suggest that these effects of Ran and Cim are mainly attributed to cardiac H2 receptor blockade.

Animals↗
PubMed1989

[Effects of ranitidine and cimetidine on ventricular fibrillation threshold and dispersion of refractory period in early myocardial ischemia].

The effects of two H2-receptor antagonists, ranitidine (Ran) and cimetidine (Cim), on the ventricular fibrillation threshold (VFT) and dispersion of the refractory period in animals with early acute myocardial ischemia induced by coronary artery ligation (CAL) were studied. The measurement of VFT was obtained in anaesthetized rats. The control group showed a decrease in VFT from 6.7 +/- 0.4 to 3.1 +/- 0.7 V 5 min after CAL, while Ran (15 mg/kg iv) and Cim (40 mg/kg iv) increased VFT from 2.8 +/- 0.7 to 5.4 +/- 1.7 V and from 3.1 +/- 0.8 to 8.1 +/- 2.7 V, respectively. By means of suction electrodes, FRP in different ischemic zones were recorded in anaesthetized rabbits, and differences between FRP in different zones were taken as the dispersion of FRP. After CAL, the control group exhibited a prolongation of FRP in central ischemic zone and a shortening of FRP in boundary zone, i.e., a dispersion of FRP was increased. Both Ran (5 mg/kg iv) and Cim (25 mg/kg iv) markedly decreased the extent of dispersion of FRP. It is concluded that the anti-arrhythmic effects of Ran and Cim may be attributable to increases in VFT and decreases in dispersion of FRP.

Animals↗
PubMed1989

Anti-arrhythmic action of cycloprotobuxine-A.

Cycloprotobuxine-A (CPB-A) 1-4 mg/kg (1/100-1/25 LD50) produced therapeutic and prophylactic effects which were found to be dose-dependent on experimental arrhythmias induced by BaCl2, aconitine and chloroform. Given at equitoxic doses, the anti-arrhythmic action of CPB-A was as potent as cyclovirobuxine-D (CVB-D) and amiodarone (Amio). However, its therapeutic index (LD50/ED50) was 1.8 times that of CVB-D and 1.2 times that of Amio. The most pronounced effects of CPB-A (0.3-30 mumol/L) on the electrophysiology of ventricular muscle of guinea pig were the lengthening of APD50, APD90 and ERP. This may contribute to its anti-arrhythmic action and suggests that CPB-A most likely belongs to class III anti-arrhythmic drugs (prolongation of APD). Perfused with the same concentration (3 mumol/L), CPB-A brought about more significant increases in APD50, APD90 and ERP than CVB-D and Amio did.

Action Potentials↗
PubMed1989

Positive inotropic effect of cycloprotobuxine-A on isolated guinea pig myocardium.

Cycloprotobuxine-A (CPB-A) 0.1-100 mumoles/L produced positive inotropic effects in left and right atria in a concentration-dependent manner. CPB-A 30 mumoles/L enhanced post-rest contraction, augmented the response of left atrium to increase in stimulating frequency, and increased the developing tension evoked by paired pulse stimulation. By taking simultaneous recordings of action potentials and contractile force of papillary muscles, it was found that CPB-A 30 mumoles/L increased the contractile force and prolonged the action potential duration at 50% of depolarization. It is concluded that the positive inotropic effect of CPB-A on myocardium may be associated with an increase in transsarcolemmal influx of calcium as well as an augmentation of the amount of calcium released from intracellular stores.

Action Potentials↗