No abstract available.
Blood Platelets*

No abstract available.
Heart Valves* ; Heart* ; Return to Work*

Human caspase-2, Ich-1 (Ice and Ced-3 homolog), has two different forms of mRNA species derived from alternative splicing, which encodes Ich-1 and Ich-1s. Ich-1v which induces apoptosis is antagonist of Ich-1s which suppresses Rat-1 cell death by serum deprivation. To investigate functions of Ich-1 and Ich-1s in T celi apoptosis, the fusion DNA constructs were made with the ecto and transmembrane of CDB and Ich-lv or Ich-1s and CDS-Ich-1 or CD8-Ich-1s chimeric protein was transiently expressed on Jurkat T cells. Tyrosine phosphorylation of intracellular proteins was induced in these transfectans when activated shortly by anti-CDB Ab. CDB-Ich-li transfectant in serum-rich condition and CDB-Ich-ls transfectant in serum-deprived condition underwent apoptosis when treated with anti-CDS Ab or incubated with NIH3T3 cells expressing stably Fas-L on their surface. We also made six antisense DNA constructs which could specifically inhibit the expression of Ich-1v, Ich- 1s, and then they were transiently transfected into Jurkat T cell. The overexpression of both of the antisese- Ich-1 against N-terminal 42 bp and against C-terminal 366 bp inhibited apoptosis through Fas signalling. But, when three different forms of antisense-Ich-1s were overexpressed in their transfectants, antisense-DNA against N-terminal 197 bp increased knd the one against C-terminal 66 bp inhibited apoptosis, instead the full size of antisense-DNA did not give any effects on apoptosis through Fas pathway.
Humans

No abstract available.
Plasma Exchange* ; Plasma* ; Seoul*

No abstract available.
Plasma Exchange* ; Plasma* ; Seoul*

Widely used tests for the detection of platelet antibodies in Korea include platelet suspension immunofluorescence test(PSIFT), enzyme immunoassay and mixed passive hemagglutination(MPHA). In these tests, removal of HLA antigens from platelet are required to detect platelet-specific antibodies. Modified antigen capture ELISA(MACE) is known to be very sensitive for the detection of platelet-specific antibodies, in which specific platelet glycoprotein, captured by the monoclonal antibody is used as a target antigen. MACE is very useful for the detection of platelet-specific alloantibodies in neonatal alloimmune thrombocytopenia(NAIT) and posttransfusion purpura(PTP). We employed MACE in our laboratory, using AP2(anti-GPIIb/IIIa, monoclonal), #30 sera(anti-PlA1), 90-545 sera(anti-HLA-B51+52) and LYS sera(multispecific HLA antibodies). LYS sera had been used as our positive control( 1:120) in MPHA. Platelet from PIA1(+), HLA-B5 I, blood group O healthy male donor, gave positive result with #30 sera(1:40) and negative result with 90-545 sera in MACE. With LYS sera, MACE showed negative in 1:120, but positive in 1:20. So LYS sera was thought to contain strong multispecific HLA antibodies and relatively weak antibody(-ies) reacting with GPllb/Illa. Further studies employing different monoclonal antibodies, such as anti-GPIb/IX, -GPIa/Ila and -GPIV are under way.
Antibodies* ; Antibodies, Monoclonal ; Blood Platelets ; Fluorescent Antibody Technique ; Glycoproteins ; HLA Antigens ; Humans ; Immunoenzyme Techniques ; Isoantibodies ; Korea ; Male ; Tissue Donors

The cryopreservation of Red Blood Cells has many advantages of which the most important one is that it can be stored for a long period. However, in Korea, Research regarding frozen blood is still in its early stage. We evaluated the effects of transfusion of the frozen-thawed RBCs in dogs. The whole bloods were collected from 5 dogs, and the packed RBCs were obtained by centrifugation method. We made the frozen RBCs by using 40% glycerol method and stored it in -80 degrees C refrigerate for 1 month. The frozen RBCs were thawed in the 37 degrees C water bath and washed by Cell washer according to the standard protocol, and evaluated the status of them being compared with that of the unfrozen. The majorirty of the results were satisfactory to the allowable limit except high plasma hemoglobin and potassium. The frozen-thawed bloods were transfused to the two dogs and carefully observed the effects and its complications. The results were that the average value of the hemoglobin was elevated about 0.6g/dL more after transfusion than before, and there were no significant complication related to the transfusion. Thus, The frozen thawed blood transfusions in case of the experiment with dogs were proved to be safe and as effective as fresh blood, and The above method appeared to be feasible to human blood.
Animals ; Baths ; Blood Transfusion ; Centrifugation ; Cryopreservation ; Dogs* ; Erythrocytes ; Glycerol ; Humans ; Korea ; Plasma ; Potassium ; Water

BACKGROUND: Polyvinyl (PVC) plastic container plasticized with di-(2-ethylhexyl) phthalate (DEHP) has been used for the storage of platelet concentrates for five days in Korea. Authors evaluated a second generation platelet storage container plasticized with tri (2-ethylhexyl) phthalate (TOTM) which was recently produced by Green Cross Medical Corp.(Korea). METHODS: 30 units of platelet concentrates were stored in TOTM-PVC container at 22'C in a flatbed agitator. Samples were taken at day 1,3,5, and 7 from the containers and tested for platelet count, MPV, PDW, pH, HCO3-,LDH, lactate, hypotonic shock response and beta-thromboglobulin (beta-TG). Electron microscopic examination was also performed. RESULTS: The number and functions of platelets were well preserved during storage. pH was maintained above 6.8 and any evidence for platelet activation was minimal. CONCLUSION: The TOTM-PVC second generation platelet storage container recently produced by the Green Cross Medical Corp.(Korea) was able to preserve platelets for at least five days without significant storage lesions.
beta-Thromboglobulin ; Blood Platelets* ; Dihydroergotamine ; Hydrogen-Ion Concentration ; Korea ; Lactic Acid ; Osmotic Pressure ; Plastics ; Platelet Activation ; Platelet Count ; Polyvinyls

The T cell antigen receptor (TcR) in combination with costimulatory signals triggered by accessory molecules present on the surface of the antigen-presenting cells (APC) regulates the activation and growth of T lymphocytes. Calyculin A and Okadaic acid is known to be an inhibitor of serine/threonine phosphatase and RK-682 specifically blocks functions of tyrosine phosphatase. To investigate roles of these inhibitors in TcR-mediated signaling cascade, chimeric molecule CD8-5 which contains the extracellular and transmembrane domains of the human CD8a molecule and the cytoplasmic tail of TcR 5 chain were stably expressed in Jurkat cell line. CD8-5 chimeric protein induced tyrosine phosphorylation of various cytoplasmic substrates and IL-2 gene expression in a NFAT dependent manner by stimulation with anti-CD8 mAb OKT8 as seen in TcR stimulation. When CD8-5 transfectants were preincubated with Okadaic acid, Calyculin or RK682, they differentially affected tyrosine phosphorylation of signaling mediators including CD8-5 molecule. When Jurkat Tag cell line was used where SV40 T antigen is stably expressed and the expression of p-galactosidase is driven by the multiple NFAT binding sites plus minimal IL-2 promoter, these phosphatase inhibitors -RK682, Calyculin A, Okadaic acid- effectively inhibited IL-2 gene expression at the concentration of 1.2832 x 10 ' M, 3.9924 x 10 M, 7.2707 x 10 M respectively. These results suggested that Okadaic acid, Calyculin or RK682 modulate TcR-proximal as well as TcR-distal signaling events during T cell activation.
Antigen-Presenting Cells ; Antigens, Viral, Tumor ; Binding Sites ; Cell Line ; Cytoplasm ; Gene Expression ; Humans ; Interleukin-2 ; Jurkat Cells ; Okadaic Acid* ; Phosphorylation ; Receptors, Antigen, T-Cell ; T-Lymphocytes ; Tyrosine

We encountered a case of neonatal altoimmune thrombocytopenia(NAIT) due to anti-HLA-B7+B60+B61. Bilateral cephal hematoma and umbilical hematoma were noted at the time of birth. Purpura developed at the third day. Platelet count was 110,000 at birth and decreased to 66,000/micro liter at the day 4. Prothrombin time and partial prothrombin time were within normal limit. The mother's platelet count was 220,000/micro liter and she had no history of abnormal bleeding. Platelet antibody tests empolying mixed passive hemagglutination and immunofluorescence revealed that the mother's serum was reactive against the platelets from the father and the neonate, but was not reactive with her own platelets. Platelets from eight volunteer group 0 donors were tested with the mother's serum; seven were reactive and one was negative. The positive reactions were lost after chloroquine treatment of platelets. Antigen capture ELISA(ACE) and modified antigen capture ELISA employing monoclonal antibodies against platelet glycoproteins In, IIa, IIb, and IIIa were negative. Mother's serum was tested for lymphocytotoxicity against 49 donor ]ymphocytes and the specificity was found to be anti-HLA-B7+B60+B61. At the 9th day, one unit of platelet concentrate from the mother was transfused and the platelet count of the neonate rose up to 340,000/micro liter. The neonate was discharged at the day of sixteenth and the platelet count remained high thereafter.
Antibodies, Monoclonal ; Blood Platelets ; Chloroquine ; Enzyme-Linked Immunosorbent Assay ; Fathers ; Fluorescent Antibody Technique ; Hemagglutination ; Hematoma ; Hemorrhage ; Humans ; Infant, Newborn ; Mothers ; Parturition ; Platelet Count ; Platelet Membrane Glycoproteins ; Prothrombin Time ; Purpura ; Sensitivity and Specificity ; Thrombocytopenia* ; Tissue Donors ; Volunteers