BACKGROUND: Several studies have demonstrated that quantitation of hepatitis B virus (HBV) DNA in sera of HBsAg-positive patients is more useful test for the assessment of infectivity and for the evaluation of disease status than previously utilized numerous serological markers and qualitative polymerase chain reaction for the detection of HBV DNA. We tried to measure serum HBV DNA using a branched DNA (bDNA) signal amplification assay, which is recently introduced and known to be a simple and nonradioisotopic method. METHODS: Total forty patients with HBsAg were randomly selected and serum HBV DNA was measured with duplication using bDNA signal amplification assay (QUANTIPLEXTM HBV DNA ASSAY, Chiron, USA). Quantitation was determined from a standard curve and expressed as HBV DNA equivalents/mL (Eq/mL; 1 Eq = 1 molecule of the primary HBV DNA standard). Serum HBeAg, aspartate aminotransferase (AST), alanine aminotransferase (ALT) , and soluble interleukin-2 receptor (sIL-2R) were compared with HBV DNA. RESULTS: Serum HBV DNA was quantitated in 13 patients (32.5%) (range 6.4x106-7.4x109 Eq/mL, mean 1.8x109 Eq/mL, CV 8.1%). All eleven patients (100%) with both HBsAg and HBeAg an4 2 of 29 patients (6.9%) with HBsAg but not with HBeAg showed measurable HBV DNA (p < 0.001). In addition, serum levels of AST, ALT, and sIL-2R were significantly higher in HBV DNA measured patients compared with those of unmeasured patients. CONCLUSIONS: Above results show that more than half the HBsAg-positive patients do not have enough HBV DNA which is measurable with boNA signal amplification assay but all of HBeAg-positive patients and some of HBeAg-negative patients do. In addition, HBV DNA quantitation might be correlated with the disease activity in HBsAg-positive patients because serum levels of AST, ALT, and sIL-2R are higher in patients measured with HBV DNA than unmeasured.
Alanine Transaminase ; Aspartate Aminotransferases ; Branched DNA Signal Amplification Assay* ; DNA ; Hepatitis B e Antigens ; Hepatitis B Surface Antigens ; Hepatitis B virus* ; Hepatitis B* ; Hepatitis* ; Humans ; Interleukin-2 ; Polymerase Chain Reaction

No abstract available.
Oligoclonal Bands*

BACKGROUND: T cell mediated immune destruction is an important mechanism of liver injury in patients with chronic hepatitis C. Serum levels of soluble interleukin-2 receptor(sIL-2R) seem to serve as a marker for the T cell activation and progressive liver injury, This study examined serum levels of sIft-2R and hepatitis C virus (HCV) RNA in patients with chronic HCV infection to determine the correlation with the severity of chronic hepatocellular damage. METHODS: Serum levels of sIft-2R in 73 patients with HCV infection (chronic hepatitis 52, liver cirrhosis 9, hepatocellular carcinoma 12) and 40 healthy controls were measured by sandwich enzyme immunoassay (CELLFREE, T Cell Sciences, USA). HCV RNA was quantified by QUANTIPLEX(TM) HCV RNA 2.0 assay (Chiron, USA) with duplication. This assay is a sandwich nucleic acid hybridization procedure using branched DNA amplification for the quantitation of HCV RNA. RESULTS: The sIL-2R levels of 52 patients with chronic hepatitis (591.4+/-238.7U/mL), 9 with liver cirrhosis(949.4+/-721.9 U/mL), and 12 with hepatocellular carcinoma (1,167.4+/- 554.4 U/mL) were significantly higher than those of healthy controls(370.8+/-71.8 U/mL) (p<0.001). A progressive and significant increase occurred in sIL-2R levels with chronic hepatitis C, liver cirrhosis and hepatocellular carcinoma (HCC) in order (p(0.001). The HCV RNA was detected in all patients and the means of HCV viral load were 3.3 MEq/mL in chronic hepatitis, 2.8 MEq/mL in cirrhosis, and 3.7 MEq/mL in HCC. There was no significant correlation between HCV RNA and the severity of liver injury in chronic HCV infection. There were no correlations among sIL-2R, HCV RNA and serum ALT. CONCLUSIONS: These results suggest that chronic hepatocellular injury by HCV progress mainly by T cell mediated immune response, not by direct cytopathic injury. Also, sIL-2R can be useful as a marker in monitoring the patients with HCV infection at high risk of getting HCC.
Carcinoma, Hepatocellular ; DNA ; Fibrosis ; Hepacivirus* ; Hepatitis C* ; Hepatitis C, Chronic* ; Hepatitis* ; Hepatitis, Chronic* ; Humans ; Immunoenzyme Techniques ; Interleukin-2* ; Liver ; Liver Cirrhosis ; Nucleic Acid Hybridization ; RNA ; Viral Load

BACKGROUND: T cell mediated immune destruction is an important mechanism of liver injury in patients with chronic hepatitis C. Serum levels of soluble interleukin-2 receptor(sIL-2R) seem to serve as a marker for the T cell activation and progressive liver injury, This study examined serum levels of sIft-2R and hepatitis C virus (HCV) RNA in patients with chronic HCV infection to determine the correlation with the severity of chronic hepatocellular damage. METHODS: Serum levels of sIft-2R in 73 patients with HCV infection (chronic hepatitis 52, liver cirrhosis 9, hepatocellular carcinoma 12) and 40 healthy controls were measured by sandwich enzyme immunoassay (CELLFREE, T Cell Sciences, USA). HCV RNA was quantified by QUANTIPLEX(TM) HCV RNA 2.0 assay (Chiron, USA) with duplication. This assay is a sandwich nucleic acid hybridization procedure using branched DNA amplification for the quantitation of HCV RNA. RESULTS: The sIL-2R levels of 52 patients with chronic hepatitis (591.4+/-238.7U/mL), 9 with liver cirrhosis(949.4+/-721.9 U/mL), and 12 with hepatocellular carcinoma (1,167.4+/- 554.4 U/mL) were significantly higher than those of healthy controls(370.8+/-71.8 U/mL) (p<0.001). A progressive and significant increase occurred in sIL-2R levels with chronic hepatitis C, liver cirrhosis and hepatocellular carcinoma (HCC) in order (p(0.001). The HCV RNA was detected in all patients and the means of HCV viral load were 3.3 MEq/mL in chronic hepatitis, 2.8 MEq/mL in cirrhosis, and 3.7 MEq/mL in HCC. There was no significant correlation between HCV RNA and the severity of liver injury in chronic HCV infection. There were no correlations among sIL-2R, HCV RNA and serum ALT. CONCLUSIONS: These results suggest that chronic hepatocellular injury by HCV progress mainly by T cell mediated immune response, not by direct cytopathic injury. Also, sIL-2R can be useful as a marker in monitoring the patients with HCV infection at high risk of getting HCC.
Carcinoma, Hepatocellular ; DNA ; Fibrosis ; Hepacivirus* ; Hepatitis C* ; Hepatitis C, Chronic* ; Hepatitis* ; Hepatitis, Chronic* ; Humans ; Immunoenzyme Techniques ; Interleukin-2* ; Liver ; Liver Cirrhosis ; Nucleic Acid Hybridization ; RNA ; Viral Load

No abstract available.
Humans ; von Willebrand Disease, Type 1* ; von Willebrand Diseases

No abstract available.
Humans ; von Willebrand Disease, Type 1* ; von Willebrand Diseases

BACKGROUND: Growth retardation (GR) has literally hundreds of causes that have differences in prognoses, complications, and responses to treatments. Especially, growth retarded patients resulting from chromosomal disorders should be genetically counseled. To focus the cytogeneticist's attention on specific chromosomal regions, it is important to understand cytogenetic aberrations associated with GR prior to chromosomal analysis. So, we attempted to figure out the cytogenetic findings of patients with GR and support the effective application of cytogenetic studies, accurate diagnosis and genetic counseling. METHODS: Of 203 cases referred for GR, the positive rate and pattern of chromosomal aberrations were retrospectively analyzed with review of associated clinical features. Cytogenetic studies were performd by high resolution banding technique after peripheral T lymphocyte culture and, if necessary, Ag-NOR stain, C-banding and fluorescence in situ hybridization. RESULTS: Forty two (20.7%) patients had abnormal karyotypes. Thirty one (15.2%) patients had well-recognized chromosomal syndromes including Turner, Cri-du-chat, Edward, 13q- and 18p-/ 18q- syndromes. In addition to those, the sporadic chromosomal aberrations of 11 cases were partial monosomy of 11q23, partial trisomy of 1q32, 4p, 9p, and 14q, and ring chromosome 4 and 18, etc. which were not literally established in view of the correlation with phenotype. CONCLUSIONS: The various results of definite or debating chromosomal disorders associated with GR could be used as the data for diagnosis, management, prognosis, and genetic counseling in growth retarded patients. Furthermore, these may provide the background for prospective study to define the new chromosomal disorders.
Abnormal Karyotype ; Chromosome Aberrations* ; Chromosome Deletion ; Chromosome Disorders ; Cytogenetics* ; Diagnosis ; Fluorescence ; Genetic Counseling ; Humans ; In Situ Hybridization ; Lymphocytes ; Phenotype ; Prognosis ; Retrospective Studies ; Ring Chromosomes ; Trisomy

BACKGROUND: As a result of exposure to human leukocyte antigen(HLA) by pregnancy, blood transfusion and previous organ transplantation, many patients awaiting renal transplantation can develop HLA antibodies. The level of HLA sensitization is determined by PRA(panel reactive antibody) test using a lymphocyte panel from HLA phenotyped selected donors. In Korea, PRA tests have not been performed routinely for organ transplantations. and there is no available data about HLA sensitization in renal transplantation. METHODS: PRA test was done in 136 sera of chronic renal failure(CRF) patients receiving dialysis (hemodialysis 108, peritoneal dialysis 28) by NIH standard microlymphocytotoxicity method with a frozen lymphocytes panel from 36 HLA-typed donors. PRA positive sera were re-tested after dithiothreitol(DTT) treatment and analyzed for HLA antibody specificities. RESULTS: Thirty five out of 136 sera(25.7%) showed positive PRA values in HLA antibody screening test. The PRA(%) values of the 35 positive sera were distributed into 1-10%(n=8), 10-20%(n=7), 20-50%(n=12) and 50%-100%(n=8). respectively. After DTT treatment, the change of PRA reactivity was divided into three groups. The PRA values of Group A(22 sera: 63%) showed no change, Group B(7 sera: 20%) declined, and Group C(6 sera. 17%) completely disappeared after DTT treatment. The specificities of HLA antibodies were identified in 19 out of 35 sera(54%). The success rate in defining antibody specificities was 0 at PRA values of 1-10% and 70-100%, and high at PRA values of 20-70%. CONCLUSION: We observed that about a quarter of CRF patients have developed HLA antibodies of immunoglobulin class-IgG, mixed IgG and IgM, and IgM HLA antibody in decreasing order of frequency.
Antibodies ; Antibody Specificity ; Blood Transfusion ; Dialysis ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Immunoglobulins ; Isoantibodies* ; Kidney Failure, Chronic* ; Kidney Transplantation ; Korea ; Leukocytes ; Lymphocytes ; Mass Screening ; Organ Transplantation ; Peritoneal Dialysis ; Pregnancy ; Tissue Donors ; Transplants

BACKGROUND: In pregnancy, paternal human leukocyte antigen (HLA) that the fetus possesses can induce the development of cytotoxic HLA antibodies in the pregnant women. We investigated the frequency and the characteristics of HLA antibodies during the pregnancy in Koreans. METHODS: Sera from 192 pregnant women (46 in the 1st trimester, 120 in the 2nd and 26 in the 3rd trimester) were tested for the presence of HLA antibody. Home made lymphocyte panel from 36 volunteers whose HLA-A, B and C antigens had been already identified 3nd formerly frozen in a liquid nitrogen tank were dispensed in duplicate into 72-well microplates and used as testing trays Test sera of one pregnant women with one negative control serum were dispensed in each plate and the plates were tested by microlymphocytotoxic method using anti-human immunoglobulin. The results were observed under fluorescence microscope and PRA (panel reactive antibody) values were determined by the percentage of wells showing positive reactions. HLA antibody specificities were identified by analysis of reaction characteristics. RESULTS: Among the 192 sera, 22 (11.5%) showed positive PRA value (PRA > 0%) in HLA antibody screening tests, in which 20 were less than 50% and 2 were more than 50% of PRA value. Two of the 46 subjects (4.3%) in the 1st trimester, 15 of the 120 (12.5%) in the 2nd and 5 of the 26 (19 2%) in the 3rd trimester were positive for HLA antibody. Among the 22 positive sera , specificities of HLA antibodies were identified in 14 (64%) sera: 8 sera had HLA antibody against single private HLA antigen. 5 had HLA antibodies against two or more antigens. and 1 sera showed anti-Bw4 antibody. CONCLUSION: In 192 pregnant women, 22 (11.5%) had HLA-A, B antibodies and they showed higher frequencies with the progress of pregnancy. Most of the pregnant women who were positive for HLA antibodies showed PRA value less than 50%. HLA antibody specificities were identified in 14 out of 22 positive sera (64%).
Antibodies* ; Antibody Specificity ; Female ; Fetus ; Fluorescence ; HLA-A Antigens* ; Humans ; Immunoglobulins ; Leukocytes ; Lymphocytes ; Mass Screening ; Nitrogen ; Pregnancy ; Pregnant Women* ; Volunteers

BACKGROUND: CD34 positive cell enumeration by flow cytometry is currently used to determine the optimal timing of peripheral blood stem cell collections (PBSC) and to predict engraftment of stem cell transplantation. However, the technical problems and lack of a standardized method are sources of significant variability in the quantitation of the CD34 positive cells. ProCOUNT(TM) (Beckon Dickinson Immuno- cytometry System, USA) kit for three color flow cytometric analysis was introduced to enumerate CD34 positive cells using a standardized method. This study was conducted to evaluate the usefulness of the three color method, ProCOUNT(TM), in comparison with two color method. METHODS: CD34 positive cells from 25 cord blood samples were enumerated by two methods, two color (CD34-PE/CD45-FITC) and three color (ProCOUNT(TM) , nucleic acid dye/CD34-PE/ CD45-PerCP) flow cytometric analysis, in which CD34 positive cells were counted directly in comparison with counting beads introduced in the sample. RESULTS: The count of CD34 positive cells in the cord blood was 28.3(+/-20.0)/uL and 20.9 (+/-16.0) /uL by three color and two color methods, respectively, The number of CD34 positive cells enumerated by ProCOUNTTM kit was well correlated with that by two color method, but the count was significantly higher in the former method (p<0.01). CONCLUSIONS: In the three color method, loss of stem cells was significantly lower than that in the two color method, and it was possible to obtain a direct count of CD34 positive cells by using a standardized procedure.
Fetal Blood* ; Flow Cytometry ; Hematopoietic Stem Cells* ; Stem Cell Transplantation ; Stem Cells