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Biomedical subjects

Y C Wu

Publications and source records attributed to Y C Wu.

At least 109 records · Page 6Linked to original sources

Pharmacological evaluation of ocoteine, isolated from Cassytha filiformis, as an alpha 1-adrenoceptor antagonist in rat thoracic aorta.

Ocoteine, isolated from Cassytha filiformis, was found to be an alpha 1-adrenoceptor blocking agent in rat thoracic aorta as revealed by its competitive antagonism of phenylephrine-induced vasoconstriction (pA2 = 7.67 +/- 0.09). Removal of endothelium from the aorta did not affect its antagonistic potency (pA2 = 7.97 +/- 0.07). [3H]-Inositol monophosphate formation caused by noradrenaline (3 microM) was suppressed by ocoteine (10 microM) and prazosin (3 microM). Ocoteine did not affect the contraction induced by U-46619, prostaglandin F2 alpha or angiotensin II, but inhibited slightly those by high K+ and endothelin I. Neither the cyclic AMP nor cyclic GMP content of rat thoracic aorta was changed by ocoteine (10 microM). Comparing the EC50 values, the potency of ocoteine against 5-hydroxytryptamine (5-HT) was about 60 times less than that against phenylephrine. Ocoteine (10 microM) also slightly antagonized the clonidine-induced inhibition of the twitch response evoked by field stimulation in rat vas deferens. In guinea pig trachea, the contraction caused by carbachol, histamine, neurokinin A or leukotriene C4 and beta 2-adrenoceptor-mediated relaxing responses induced by isoprenaline were not affected by ocoteine (10 microM). The voltage clamp study in rat ventricular single myocytes revealed that ocoteine (3, 10 microM) inhibited steady state outward currents, but not transient outward currents or slow inward Ca2+ currents. It is concluded that ocoteine is a selective alpha 1-adrenoceptor antagonist in isolated rat thoracic aorta. At high concentrations, it also blocks 5-HT receptors and Na+ and steady state outward currents in rat ventricular myocytes.

Adrenergic alpha-1 Receptor Antagonists↗

[Seroepidemiology of Japanese encephalitis viral infection among 3-6 years old children from mountainous and plains townships located in the northern, central, southern and eastern Taiwan].

In order to evaluate the Japanese encephalitis virus (JEV) vaccination program in rural Taiwan, we conducted a seroepidemiological survey of JEV among rural children 3 to 6 years of age in Taiwan. The children were selected through a systemic sampling following stratification by age of children in 4 selected aboriginal villages and 4 adjacent nonaboriginal villages. The overall vaccine coverage rate for the primary (2 doses) dose was 81.2% (1853/2281) with higher rates (87.7%-87.9%) found among the more recent birth cohort of 3 to 4 years of age. The neutralizing antibody (NT) against JEV was measured with plaque reduction neutralization test (PRNT) using Nakayama strain as the virus. With a positive NT antibody defined as > or = 1:10 dilution of serum yielding more than 50% plaque reduction, the overall JEV NT antibody positive rate among children receiving 3 doses of vaccine was 67%. However, the age-specific positive rates varied significantly with varying ages; the lowest of 47% being among children 4 years of age which was lower than the rates of 68%, 76% and 87% among children of 3, 5 and 6 years of age, respectively. This trend of rising seropositive rates of JEV antibody with increasing age among 4 and 6 years of age was also noted among children who had received no vaccine, suggesting the importance of natural infection among rural Taiwanese children. Despite the high frequency of natural infection, the seropositive rates of JEV antibody still correlated well with the dose of vaccine received, i.e., 67% (1122/1664), 66% (65/97), 33% (4/12) and 40% (19/47) for children receiving 3, 2, 1, and 0 dose of JE vaccines, respectively (P < 0.0001 Chi-square for trend test). When stratified analysis by dose and by type of vaccines was conducted, a significantly higher seropositive rate of JEV NT antibody was noted among children receiving JE vaccine of Beijing type (87%) than children receiving Nakayama type (39%) (p < 0.0001, Chi-square test). Our data indicated that the JEV vaccination, in conjunction with JEV natural infection, has maintained high JEV NT antibody level among rural children of Taiwan.

Age Factors↗

[Detection of neutralizing antibodies to Japanese encephalitis virus by enzyme-linked immunosorbent assay (ELISA)].

Competitive ELISA was used for the detection of neutralizing antibody to JE. Based on the principle that human serum JE antibody competed with JE monoclonal antibody (MAb) for JE antigen, it was found that 3 JE MAbs (E3-3, NPF-5 and NNN-5) were suitable for competitive ELISA for the detection of JE neutralizing antibody. The sensitivity of cometitive ELISA for 29 JE confirmed serum specimens with titer of plaque reduction neutralization test (PRNT) was checked to be 82.1% (23/28). The specificity of E3-3 MAb to JE used in competitive ELISA was 100%. Correlation coefficient of JE confirmed cases of 57 hemagglutination inhibition (HI) titers in 1995 and 37 PRNT titers in 1994 compared with competitive ELISA were 0.744 and 0.732, respectively. Compared the competitive ELISA titers of 154 sera of healthy people with PRNT titers, the results showed that 70% of the sera could be detected by competitive ELISA which saved a lot of time and manpower.

Antibodies, Viral↗

Delayed clearance of transmitter and the role of glutamate transporters at synapses with multiple release sites.

The roles of glutamate diffusion, uptake, and channel kinetics in shaping the AMPA receptor EPSC were examined at a calyceal synapse. The EPSC decay was described by three exponential components: two matching desensitizing channel kinetics, and a third component at least 10 times slower. The slowest component had identical voltage dependence to the steady-state AMPA current and was selectively increased and prolonged by blockade of glutamate uptake, indicating that the slow EPSC represented rebinding of glutamate at partially desensitized AMPA receptors. The data were in strong agreement with the predictions of a model of transmitter diffusion from multiple release sites into a large synaptic cleft. Within the first millisecond after release, transmitter concentrations in the cleft fell below millimolar levels, causing the fastest parts of the EPSC to be shaped by channel kinetics. The slowest component was determined by the removal over tens of milliseconds of the final 10-100 microM glutamate by diffusion and uptake. The data and modeling indicate that transmitter uptake and cooperation between release sites are significant determinants of a slow "tail" of glutamate in the synaptic cleft. This slow clearance of glutamate is likely to limit postsynaptic receptor availability through desensitization.

Animals↗

Early and advanced glycosylation end products. Kinetics of formation and clearance in peritoneal dialysis.

The chronic contact of glucose-containing dialysate and proteins results in the deposition of advanced glycation end products (AGEs) on peritoneal tissues in patients treated by peritoneal dialysis (PD), yet plasma levels of the AGE pentosidine are significantly lower in PD than in hemodialysis (HD). We measured glycation of peritoneal proteins in patients on PD over the time course of intraperitoneal equilibration of fresh peritoneal dialysate. The glycated content of peritoneal proteins (furosine method) was initially identical to plasma but increased 200% within 4 h due to in situ glycation as also demonstrated in vitro. In contrast, peritoneal proteins contained a 2-4 x greater content of the AGE pentosidine at all equilibrium time points. Plasma protein furosine content was identical in patients on PD and on HD. Fractionation by gel filtration of serum from patients on PD and HD revealed that > 95% of the pentosidine was linked to proteins > 10,000 mol wt; < 1% to proteins < 10,000 mol wt; and < 1%, free. Neither HD nor PD affected protein-bound pentosidine. The HD treatment decreased free and < 10,000 mol wt bound pentosidine. However clearance of protein-associated pentosidine by the peritoneal membrane may explain lower steady state levels in patients treated by PD.

Adult↗

Identification of immunodominant, group-specific and subcomplex-specific, continuous epitopes in the core regions of Japanese encephalitis virus using synthetic peptides.

Two flaviviruses, Japanese encephalitis (JE) virus and Dengue (DEN) virus which have high pathogenicity for humans, continue to pose a serious public health problem in tropical and subtropical countries of the world. In order to identify the immunodominant B-cell epitopes for diagnostic application, we have prepared a series of 15-mer synthetic peptides from JE virus core protein based on computer analysis. Four linear, immunodominant epitopes corresponding to amino acids 91-105 (P78), 1-15 (P73), 8-22 (P74), and 34-48 (P75) of JE virus core proteins were identified by employing an enzyme-linked immunosorbent assay (ELISA), using high-titered immune sera from JE-vaccinated children. P78 was found to be the most immunodominant. The sero-specificity of these peptides was tested by binding to seroconverted samples from JE and DEN-1 patients. P78 and P74 belonged to group-specific epitopes which reacted with both JE and DEN-1 patient sera. P73 and P75 belonged to subcomplex-specific epitopes which reacted only with JE but not with DEN-1 patient sera. The study suggests that these peptides corresponding to the immunodominant epitopes of JE virus core protein might have the potential to be used as peptide-based diagnostic reagents for the detection and differentiation of JE and DEN antibody responses.

Adult↗

A theoretical study of calcium microdomains in turtle hair cells.

Confocal imaging has revealed microdomains of intracellular free Ca2+ in turtle hair cells evoked by depolarizing pulses and has delineated factors affecting the growth and dissipation of such domains. However, imaging experiments have limited spatial and temporal resolution. To extend the range of the results we have developed a three-dimensional model of Ca2+ diffusion in a cylindrical hair cell, allowing part of the Ca2+ influx to occur over a small circular region (radius 0.125-1.0 micron) representing a high-density array of voltage-dependent channels. The model incorporated experimental information about the number of channels, the fixed and mobile Ca2+ buffers, and the Ca2+ extrusion mechanism. A feature of the calculations was the use of a variable grid size depending on the proximity to the Ca2+ channel cluster. The results agreed qualitatively with experimental data on the localization of the Ca2+ transients, although the experimental responses were smaller and slower, which is most likely due to temporal and spatial averaging in the imaging. The model made predictions about 1) the optimal Ca2+ channel number and density within a cluster, 2) the conditions to ensure independence of neighboring clusters, and 3) the influence of the Ca2+ buffers on the kinetics and localization of the microdomains. We suggest that an increase in the mobile Ca2+ buffer concentration in high-frequency hair cells (which possess a larger number of release sites) would allow lower amplitude and faster Ca2+ responses and promote functional independence of the sites.

Animals↗

A developmental model for generating frequency maps in the reptilian and avian cochleas.

Hair cells in the turtle cochlea are frequency-tuned by a mechanism involving the combined activation of voltage-sensitive Ca2+ channels and Ca(2+)-activated K+ (KCa) channels. The main determinants of a hair cell's characteristic frequency (Fo) are the KCa channels' density and kinetics, both of which change systematically with location in the cochlea in conjunction with the observed frequency map. We have developed a model based on the differential expression of two KCa channel subunits, which when accompanied by concurrent changes in other properties (e.g., density of Ca2+ channels and inwardly rectifying K+ channels), will generate sharp tuning at frequencies from 40 to 600 Hz. The kinetic properties of the two subunits were derived from previous single-channel analysis, and it was assumed that the subunits (A and B) combine to form five species of tetrameric channel (A4, A3B, A2B2, AB3, and B4) with intermediate kinetics and overlapping distribution. Expression of KCa and other channels was assumed to be regulated by diffusional gradients in either one or two chemicals. The results are consistent with both current- and voltage-clamp data on turtle hair cells, and they show that five channel species are sufficient to produce smooth changes in both Fo and kinetics of the macroscopic KCa current. Other schemes for varying KCa channel kinetics are examined, including one that allows extension of the model to the chick cochlea to produce hair cells with Fo's from 130 to 4000 Hz. A necessary assumption in all models is a gradient in the values of the parameters identified with the cell's cytoplasmic Ca2+ buffer.

Animals↗

Antiplatelet of vasorelaxing actions of some benzylisoquinoline and phenanthrene alkaloids.

Members of a series of benzylisoquinoline and phenanthrene alkaloids were tested for their antiplatelet and vasorelaxing actions. Atherosperminine HClO4 (15) and atherosperminium I (16) showed strong inhibition of adenosine 5'-diphosphate-induced platelet aggregation. dl-N-Methylcoclaurine (1), dl-coclaurine (6), 15, 16, xylopine hydroxylamine (18), and atherosperminine N-oxide (19) showed strong inhibition of arachidonic acid-induced platelet aggregation. Compounds 1, 15, 16, dicentrine methine (17), 18, and 19 showed strong inhibition of collagen-induced platelet aggregation. d-(-)-Magnocurarine I (8), 15, and 16 showed strong inhibition of platelet-activating factor-induced platelet aggregation. Compounds 15, 17, and 19 showed vasorelaxing action in rat thoracic aorta.

Alkaloids↗

Identification of ent-16 beta, 17-dihydroxykauran-19-oic acid as an anti-HIV principle and isolation of the new diterpenoids annosquamosins A and B from Annona squamosa.

Phytochemical analysis of the fruits of Annona squamosa yielded 12 known kaurane derivatives (1-11, 13) and two new kaurane diterpenoids, which have been named annosquamosin A (16 beta-hydroxy-17-acetoxy-ent-kauran-19-al) (12) and annosquamosin B (19-nor-ent-kaurane-4 alpha,16 beta,-17-triol) (14). The structures of the new compounds were established by spectral analyses and chemical evidence. Among these 14 compounds, 16 beta, 17-dihydroxy-ent-kauran-19-oic acid (2) showed significant activity against HIV replication in H9 lymphocyte cells with an EC50 value of 0.8 microgram/mL (therapeutic index > 5).

Antiviral Agents↗

Antiplatelet and vasorelaxing actions of some aporphinoids.

A series of aporphines and oxoaporphines was tested for antiplatelet and vasorelaxing actions. (+)-N-Methylactinodaphnine, (-)-norannuradhapurine x HBr, xylopine, actinodaphnine, and N-methylnandigerine showed strong inhibition of adenosine 5'-diphosphate (ADP)-induced platelet aggregation. Boldine, (+)-N-methylactinodaphnine, (-)-norannuradhapurine x HBr, xylopine, N-acetyllaurolitsine, N-methyllaurotetanine, actinodaphnine, N-methylnandigerine, O-methylbulbocapnine, and liriodenine showed strong inhibition of arachidonic acid (AA)-induced platelet aggregation. (+)-N-Methylactinodaphnine, fissoldine x HCIO4, (-)-norannuradhapurine x HBr, xlylopine, N-methyllaurotetanine, actinodaphnine, N-methylnandigerine, O-methylbulbocapnine, and liriodenine showed strong inhibiton of collagen-induced platelet aggregation. (-)-Norannuradhapurine x HBr, xylopine, N-methyllaurotetanine, and actinodaphnine showed strong inhibition of platelet-activating factor (PAF; 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine)-induced platelet aggregation. (+)-N-Methylactinodaphnine, laurotetanine, N-methylactinodaphnine N-oxide, oxoglaucine, boldine, and actinodaphnine showed vasorelaxing action in rat thoracic aorta. The results are discussed on the basis of structure-activity relationships.

Adenosine Diphosphate↗

Electrophysiological mechanisms for antiarrhythmic efficacy and positive inotropy of liriodenine, a natural aporphine alkaloid from Fissistigma glaucescens.

1. The antiarrhythmic potential and electromechanical effects of liriodenine, an aporphine alkaloid isolated from the plant, Fissistigma glaucescens, were examined. 2. In the Langendorff perfused (with constant pressure) rat heart, at a concentration of 0.3 to 3 microM, liriodenine was able to convert a polymorphic ventricular tachyrhythmia induced by the ischaemia-reperfusion (EC50 = 0.3 microM). 3. In isolated atrial and ventricular muscle, liriodenine increased the contractile force and slowed the spontaneous beating of the right atrium. 4. The liriodenine-induced positive inotropy was markedly attenuated by a transient outward K+ channel blocker, 4-aminopyridine (4-AP) but was not significantly affected by prazosin, propranolol, verapamil or carbachol. 5. In rat isolated ventricular myocytes, liriodenine prolonged action potential duration and decreased the maximal upstroke velocity of phase 0 depolarization (Vmax) and resting membrane potential in a concentration-dependent manner. The action potential amplitude was not significantly changed. 6. Whole-cell voltage clamp study revealed that liriodenine blocked the Na+ channel (INa) concentration-dependently (IC50 = 0.7 microM) and caused a leftward shift of its steady-state inactivation curve. However, its recovery rate from the inactivated state was not affected. The L-type Ca2+ currents (Ica) were also decreased, but to a lesser degree (IC50 = 2.5 microM, maximal inhibition = 35%). 7. Liriodenine inhibited the 4-AP-sensitive transient outward current (Ito) (IC50 = 2.8 microM) and moderately accelerated its rate of decay. The block of Ito was not associated with changes in the voltage-dependence of the steady-state inactivation curve or in the process of recovery from inactivation of the current. Liriodenine also reduced the amplitude of a slowly inactivating, steady-state outward current (Iss) (IC50 = 1.9 microM). These effects were consistent with its prolonging effect on action potential duration. The inwardly rectifying background K+ current (IK1), was also decreased but to a less degree. 8. Compared to quinidine, liriodenine exerted a stronger degree of block on INa, comparable degree of block on IK1, and lesser extent of block on ICa and Ito. 9. It is concluded that, through inhibition of Na+ and the Ito channel, liriodenine can suppress ventricular arrhythmias induced by myocardial ischaemia reperfusion. The positive inotropic effect can be explained by inhibition of the Ito channel and the subsequent prolongation of action potential duration. These results provide a satisfactory therapeutic potential for the treatment of cardiac arrhythmias.

Action Potentials↗

Complete nucleotide sequence of the genome of Japanese encephalitis virus ling strain: the presence of a 25-nucleotide deletion in the 3'-nontranslated region.

The complete sequence of the genome of the Japanese encephalitis virus (JEV) Ling strain isolated from the brain of a patient in Taiwan in 1965 was cloned by using the reverse transcription-polymerase chain reaction method. Seven overlapping cDNA clones that span the entire virus genome were isolated and sequenced to determine the complete nucleotide sequence, which is 10,951 nucleotides in length. As reported for three other JEV strains (Beijing-1, SA-14, and JaOArS982), the Ling strain contains 95 nucleotides in the 5' nontranslated region (NTR), followed by a single open reading frame of 10,296 nucleotides. However, the length of the 3' NTR of JEV Ling is 560 nucleotides, 25 nucleotides shorter than that of other JEV strains sequenced to date. Comparison of nucleotide and amino acid sequences among these four JEV strains showed that nucleotide (amino acid) sequence divergence in the translated region varied from 1.25% to 3.27% (0.49-1.63%). The nucleotide (amino acid) divergences between the Ling and Beijing-1 strains were 1.25% (0.87%) and between the SA-14 and JaOArS982 strains were 1.42% (0.49%). These values are lower than those found between the Ling and SA-14 [2.44% (1.02%)] or the Ling and JaOArS982 strains [2.84% (0.93%)], as well as those between Beijing-1 and SA-14 [3.14% (1.60%)] or Beijing-1 and JaOArS982 [3.27% (1.63%)] strains. Sequence comparisons of subregions of the genomes i.e., structural genes, nonstructural genes, or individual genes, showed divergence similar to that obtained by comparing the entire sequence. It is likely that the JEV sequence divergence between two human isolates or between two mosquito isolates is lower than that between a human isolate and a mosquito isolate.

Aedes↗

The advanced glycation endproduct pentosidine and monocyte activation in uremia.

RATIONALE: Advanced glycation endproducts (AGEs) contribute to the pathogenesis of vascular complications in diabetes, aging and end-stage renal disease (ESRD). Immune abnormalities in patients with chronic renal failure and those treated by dialysis contribute to high rates of morbidity and mortality. We therefore sought a relationship between a circulating marker of immune dysfunction and plasma levels of the AGE pentosidine. METHOD: We studied non-diabetic patients with mild to advanced renal failure (n = 60), and with ESRD treated by hemodialysis (HD) (n = 44) and peritoneal dialysis (PD) (n = 19). The plasma protein content of the well characterized AGE, pentosidine was measured using HPLC. In the same samples the monocyte activation product neopterin was measured by RIA. RESULTS: Plasma levels of pentosidine and neopterin increased in parallel with the progression of renal failure. Pentosidine and neopterin were highly correlated in all patients even after adjustment for Ccr. This correlation was also present in patients with ESRD. CONCLUSION: These data suggest that the AGE pentosidine is associated with monocyte activation in renal failure, an interaction which may contribute to accelerated rates of complication and death by as yet unknown mechanisms.

Arginine↗

Detection of IgM antibody to Japanese encephalitis virus infection by enzyme-linked immunosorbent assay (ELISA).

BACKGROUND: Japanese encephalitis (JE) is an important infectious disease in Taiwan, with reported cases observed all the year around. Laboratory tests for JE consist mainly of hemagglutination inhibition (HI) test and neutralization test (NT). Commercialized enzyme-linked immunosorbent assay (ELISA) kits for detection of JE-IgM are still not available. Therefore an attempt has been made to develop a sensitive and rapid ELISA for detection of the IgM antibody to JE to serve as an indication of recent Japanese encephalitis virus infection. METHODS: Both positive and negative JE serum specimens, confirmed by HI test, were checked for IgM antibody to JE by ELISA. The optimum concentration of biotin-IgG and avidin horse-radish peroxidase conjugate used in MAC-ELISA were 1/2000 and 1/ 15000, respectively. RESULTS: From 1987 to 1989, 118 paired serum specimens of HI-confirmed JE patients were tested for JE-IgM by ELISA. The positive rate of JE-IgM was 65.7% (25/38), 73.9% (17/23), 93.5% (29/31) and 88.8% (8/9) for the consecutive first to fourth weeks after onset of the disease. The JE-IgM antibody of 17 serum specimens collected from the 5th to the 10th week after onset of the disease were 100% detected. In addition, among the 13 HI-confirmed JE cases occurring in 1994, 84.6% of the acute phase serum specimens demonstrated the JE-IgM antibody. CONCLUSIONS: About 65.7% of the JE-IgM of the acute serum specimens collected within one week after onset of the disease were detected. The JE-IgM positive rate elevated as the days from disease onset increased. Therefore the appearance of JE-IgM could be used as an indication of recent JEV infection to serve as a rapid laboratory diagnostic tool.

Adolescent↗

Disparate changes in plasma and tissue pentosidine levels after kidney and kidney-pancreas transplantation.

The advanced glycation end-product, pentosidine, was measured in plasma proteins and skin collagen before and after kidney and kidney-pancreas transplantation in order to determine the relationship between plasma and tissue levels and to characterize the pattern of change in pentosidine levels after correction of hyperglycemia and/or renal failure. The content of pentosidine in skin collagen was higher than that in plasma proteins both before and after transplantation. However, there was no correlation between plasma and skin pentosidine levels. Prior to transplantation, the content of pentosidine in skin collagen was related to the duration of dialytic therapy, presence of diabetes mellitus, age, and female gender. Following transplantation, plasma pentosidine levels were inversely correlated with glomerular filtration rate (r = 0.64; p < 0.01). While plasma pentosidine levels consistently decreased after transplantation, levels in skin collagen increased in 10 of 13 patients, including 5 of 6 recipients of kidney-pancreas transplants. Our results indicate that tissue levels of pentosidine persist for long periods of time after kidney or kidney-pancreas transplantation, despite consistent decreases in levels measured in plasma proteins. The observed increase in tissue pentosidine levels in a majority of patients suggests that formation of advanced glycation end-products may continue after otherwise successful kidney or kidney-pancreas transplantation.

Adult↗