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Biomedical subjects

Y S Wu

Publications and source records attributed to Y S Wu.

At least 19 recordsLinked to original sources
PubMed1992

Serum independence of low K+ induction of Na,K-ATPase: possible role of c-fos.

Cultured ARL15 cells respond to abnormally low extracellular K+ concentrations by increasing the abundance of Na,K-ATPase (the Na/K pump). This response is preceded by significant increases in the mRNAs of the alpha 1 and beta 1 subunits of this enzyme, implying transcriptional or post-transcriptional regulation in the response. The present study concerned the possible participation of serum factors in low K+ induction of Na,K-ATPase. In normal K+ (4.5 mM) or low K+ (0.68 mM) the presence of 10% calf serum had no effect on Na,K-ATPase activity. The serum independence of the response to low K+ raised the possibility that low K+ may itself elicit a "growth" response. Accordingly, the effect of low K+ on mRNA abundances of four proto-oncogenes (c-fos, c-myc, c-jun and c-ski) was evaluated in the early phase of the response by quantitative Northern blot analysis. The mRNA for c-fos was transiently elevated by low K+, with a peak at 30 min. In contrast, low K+ had no measurable effect on the abundances of c-myc, c-jun and c-ski, for up to 2 hr of exposure. The early elevation of c-fos mRNA makes it a candidate mediator in this signal-transduction pathway. Induction of c-fos mRNA by the phorbol ester, PMA, or by dioctanoyl glycerol, however, had no effect on Na,K-ATPase activity. These results indicate that an increase in c-fos mRNA alone is not sufficient to induce Na,K-ATPase. Whether induction of c-fos is necessary for the response to low K+ remains to be determined in future studies.

Animals↗
PubMed1992

Mechanism of differential inhibition of lactate dehydrogenase isoenzymes in the BMC LD-1 assay.

The mechanism of inhibition of lactate dehydrogenase (LD) isoenzymes by guanidinium thiocyanate (GSCN) used in the LD-1 assay developed by Boehringer Mannheim Corporation (BMC) was investigated. Michaelis-Menten inhibition kinetics for the individual isoenzymes revealed that GSCN competitively inhibited LD-1 in the presence of lactate and NAD+, but is a noncompetitive inhibitor of LD-5. LD-2 and LD-3 exhibited mixed inhibition kinetics. The inhibition constants were two- to threefold smaller for LD-5 than for LD-1. Time-dependent studies also showed that the isoenzymes underwent a different rate of inactivation by GSCN. LD-5, LD-3, and LD-2 were rapidly inactivated within 1 min under the BMC assay conditions, whereas LD-1 lost only about 20% of activity after 10 min. The presence of lactate further protects LD-1, but not other isoenzymes. Under this condition, LD-1 was not inactivated during the initial 6 min of reaction. Separate experiments demonstrated that both guanidinium and thiocyanate ions are responsible for the inactivation process that was found to be irreversible. We speculate that GSCN selectively denatures the M subunit of LD. The H subunit is less susceptible to denaturation and is further stabilized by lactate.

Guanidines↗
PubMed1992

Evaluation of anti-HTLV-1 in physical-checkup individuals and blood donors in Taiwan.

The gelatin particle agglutination (GPA) test specific for antibody detection of human T-cell lymphotropic virus type 1 (HTLV-1) was used to screen 500 blood donors and 5000 physical-checkup individuals at Veterans General Hospital-Taipei. The positive rate of physical-checkup individuals and the blood donors was 0.18% (9/5000) and 0% (0/5000) respectively. Among the 9 GPA positive specimens, eight were confirmed to be positive by western blot analysis and a prevalence rate of 0.16% (8/5000). Seven of the nine GPA positive samples were also positive by indirect fluorescent antibody test and two of them had indeterminate results. Since GPA is less expensive, relatively simple and convenient, we recommend that GPA could be used as screening test for HTLV-1 infection of blood donors, followed by western blot method as a confirmatory test in blood bank.

Adolescent↗
PubMed1991

Leukemia initiated by hemopoietic stem cells expressing the v-abl oncogene.

We report a mouse model with which to study leukemogenesis initiated by a specific genetic change introduced into a primary lymphoid-myeloid pluripotent stem cell. Fetal liver hemopoietic cells were infected with a high titer of helper-free Abelson murine leukemia virus (A-MuLV) and were used to reconstitute lethally irradiated mice. Two weeks later, progenies of a single primitive hemopoietic stem cell carrying a specifically integrated A-MuLV proviral DNA could be detected in both colony-forming units in spleen and myeloid colony-forming cells in the bone marrow. Beginning at 3 weeks after transplantation, the recipients developed elevated leukocyte counts, splenomegaly, and increase of blast cells in the peripheral blood. Multiple clones of A-MuLV-infected cells were infused into each recipient. However, in the same animal, DNA extracted from various affected organs and from factor-independent lymphoid and myeloid immortalized cells all contained an identical, specifically integrated proviral genome. The A-MuLV-infected stem cells differentiated into various lineages of hemopoietic cells. Our data show that the expression of the v-abl oncogene in a primary lymphoid-myeloid hemopoietic stem cell directly initiates leukemogenesis by stimulating factor-independent growth. The monoclonal-type disease development seen in these animals may require the occurrence of an additional genetic event.

Abelson murine leukemia virus↗
PubMed1990

Clinical evaluation of an automated chemical inhibition assay for lactate dehydrogenase isoenzyme 1.

We evaluated an automated assay for lactate dehydrogenase (LD; EC 1.1.1.27) isoenzymes, supplied by Boehringer Mannheim Diagnostics (BMD) and based on selective chemical inhibition of non-LD-1 isoenzymes by guanidine thiocyanate. Results were compared with the Roche Isomune LD-1 method. The Hitachi 717 analyzer was used to measure enzyme activity for both procedures in 229 serum samples. One hundred specimens were also analyzed by the Helena rapid electrophoresis (REP) method. We determined the limit of linearity of the BMD method to be about 1200 U of LD-1 per liter. The analytical correlation of BMD (y) with Isomune (x) yielded y = 1.0x + 0.5 U/L, r = 0.997, Sy/x = 16.9 (range 20-1397 U/L). The regression equation for BMD vs REP was y = 1.1x + 7.2% (r = 0.800, Sy/x = 7.4, range 14-83%). Average values for within-run precision for low (38 U/L), medium (180 U/L), and high (865 U/L) controls were 4.1%, 1.0%, and 0.5%, respectively (16 trials of six each). The average values for run-to-run precision were 4.1%, 1.7%, and 1.1%, respectively, for these controls (n = 16). We used receiver-operating characteristic curves to determine optimum decision limits. Using an LD-1 cutoff of 40% of total LD, we obtained a clinical sensitivity of 97-100% and a specificity of 95% when blood was collected during the optimum interval, 24-48 h after the onset of chest pain. We conclude that the BMD LD-1 assay is equivalent to the immunochemical and electrophoretic assays for measuring the LD-1 isoenzyme.

Autoanalysis↗
PubMed1990

[Effect of 2-naphthyl-3- (3,4-dimethoxy) phenyl-propenoic acid on the metabolism of arachidonic acid in rabbit platelets].

Washed rabbit platelets were incubated with [14C] arachidonic acid (AA sodium salt) after being exposed to 2-naphthyl-3-(3,4-dimethoxy) phenyl-propenoic acid (NM-PA), indomethacin (Ind) and imidazole (Imi) or control solvent. The metabolites of AA in rabbit platelets were separated with the system of chloroform: methanol: acetic acid: water (90:8:1:0.8) by thin layer chromatography and quantitated by liquid scintillation counter. The metabolism of AA was influenced by NMPA dose-dependently in the range of 0.05-0.5 mmol/L. The formation of thromboxane B2 (TXB2) and 12-hydroxy-5,8,10-heptadecatrienoic acid (HHT), the final metabolites in the pathway of cyclooxygenase-thromboxane synthetase, were decreased from 24 +/- 6.7 to 3.2 +/- 1.6% and 10.3 +/- 2.49 to 4.7 +/- 2.8%, respectively. Meanwhile, 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE), the metabolite in the pathway of lipoxygenase, was increased from 11.9 +/- 1.7 to 34 +/- 5.6%. It is suggested that NMPA blocks the pathway of cyclooxygenase-thromboxane synthetase and then changes the direction of arachidonate metabolism in platelets, the activation of platelets is inhibited in this way.

Animals↗
PubMed1989

[Observations on substance P in the brain and spinal cord in schizophrenia].

The regional distribution of substance P in postmortem brain and spinal cord of 5 cases of schizophrenia were investigated using the immunohistochemical PAP and ABC technique. Substance P immunoreactive fibers were primarily found in Substantia nigra, nucleus tractus spinalis n. trigemini, nucleus tractus solitarii, nucleus spinalis n. vestibuli, nucleus ambiguous, Central gray matter of the midbrain, globus pallidus and cornua posteriora spinales. The 5 cases showed no differences to the controls. These results agree with the finding reported abroad that the brain tissues were measured by radioimmunoassays. It was also discussed on study method of substance P in Central nervous system.

Adolescent↗
PubMed1987

Identification of glycoprotein components of alpha-agglutinin, a cell adhesion protein from Saccharomyces cerevisiae.

Several glycoproteins which inhibit the agglutinability of Saccharomyces cerevisiae mating type a cells were partially purified from extracts of mating type alpha cells. These proteins, called alpha-agglutinin, were labeled with 125I-Bolton-Hunter reagent. The labeled alpha-agglutinin showed specific binding to a cells. Such specific binding approached saturation with respect to agglutinin or cells and was inhibited in the presence of excess unlabeled alpha-agglutinin. Nonspecific binding was similar in a and alpha cells, was neither saturable nor competable, and was three- to fourfold less than the specific binding to a cells at maximum tested agglutinin concentrations. The major a-specific binding species had a low electrophoretic mobility in sodium dodecyl sulfate-polyacrylamide gels and had an apparent molecular weight of 155,000 by rate zonal centrifugation. Endo-N-acetylglucosaminidase H digestion of the purified glycoprotein complex converted the low-mobility material to four major and several minor bands which were resolved by polyacrylamide gel electrophoresis. All but two minor peptides bound specifically to a cells. Analyses of agglutinin from mnn mutants confirmed the deglycosylation results in suggesting that the N-linked carbohydrate portion of alpha-agglutinin was not necessary for activity.

Agglutination↗
PubMed1987

Interaction of alpha-agglutinin with Saccharomyces cerevisiae a cells.

Binding of Saccharomyces cerevisiae alpha-agglutinin to target a cells was assayed by agglutination inhibition and 125I-alpha-agglutinin binding. The assays showed characteristics of equilibrium binding, namely saturability, competability, and the establishment of a kinetic endpoint in the presence of free alpha-agglutinin and free receptor. The binding was heterogeneous, displaying strong binding (10(9) liters/mol) and a weaker interaction. There were about 2 X 10(4) strong binding sites per a cell. Denaturing gels displayed identical labeled species binding to the a cells in the weak and strong interactions. Furthermore, weakly bound material could subsequently bind tightly to fresh a cells, implying that the same species of alpha-agglutinin was bound in the two states.

Agglutination↗
PubMed1978

Affinity labeling of histamine N-methyltransferase by 2',3'-dialdehyde derivatives of S-adenosylhomocysteine and S-adenosylmethionine. Kinetics of inactivation.

S-Adenosyl-L-methionine (AdoMet), S-adenosyl-L-homocysteine (L-AdoHcy), and related ribonucleosides have been oxidized with periodic acid to the corresponding 2',3'-dialdehydes. Both AdoMet dialdehyde and L-AdoHcy dialdehyde were observed to rapidly and irreversibly inactivate histamine N-methyltransferase (HMT). Equally active as an irreversible inhibitor was S-adenosyl-D-homocysteine dialdehyde (D-AdoHcy dialdehyde), which is consistent with the known affinity of HMT for S-adenosyl-D-homocysteine (D-AdoHcy). Other analogues of AdoHcy dialdehyde (S-adenosyl-L-cysteine dialdehyde, S-adenosyl-L-homocysteine sulfoxide dialdehyde, and adenosine dialdehyde) also produced irreversible inactivation of HMT, but at predictably slower rates. The corresponding acyclic 2',3'-ribonucleosides, which were obtained by NaBH4 reduction of the ribonucleosides dialdehydes, were found to be very weak, reversible inhibitors of HMT. Kinetic analysis of the inactivation of HMT produced by L-AdoHcy dialdehyde, AdoMet dialdehyde, and D-AdoHcy dialdehyde suggested mechanisms involving the formation of dissociable enzyme-inhibitor complexes prior to irreversible inactivation. Studies using L-[2,8-3H] AdoHcy dialdehyde revealed that incorporation of radioactivity into HMT closely paralleled the loss of enzyme activity. The results of these studies indicate that L-AdoHcy dialdehyde, D-AdoHcy dialdehyde, and AdoMet dialdehyde are affinity labeling reagents for HMT.

Affinity Labels↗