摘要:

AbstractUsing an automated HPLC system reported recently by us (J Clin Lab Anal4:410–413, 1990), serum unconjugated estriol and estradiol (u‐E3and u‐E2) can be measured within 22 min. The clinical value of this method was evaluated in this study with abnormal pregnancies, twin pregnancies, and pregnancies associated with fetal anencephaly or complicated by gestational toxicosis or diabetes mellitus. A significant correlation was demonstrated between an abrupt decrease in serum u‐E3level and an abnormal pattern on fetal heart rate monitor. Since urinary E3was not sensitive enough to detect any depressed fetal condition, serum u‐E3is better biochemical parameter for assessment of fetal well‐being.In addition, u‐E2was measured in the sera of nonpregnant women with separation chromatography as a pretreatment in order to obtain higher sensitivity of the system. This modified HPLC system enabled us to determine the serum u‐E2in about 60 min with the minimal detectable value of 80 pg/ml; the coefficient of correlation with RIA was 0.822. As a result, this quick measurement of u‐E3and u‐E2by our HPLC system revealed to be useful not only for evaluating fetoplacental function but also for monitoring ovarian

ISSN:0887-8013    

DOI:10.1002/jcla.1860060102

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractWe evaluated an immunoturbidimetric quantitation for serum myoglobin by the latex agglutination method using an automated biochemical analyzer. This method is rapid, specific, accurate, precise, and has wide dynamic range. The total assay time is 10 min and is performed at 37°C with continuous monitoring at 570 nm.The assay results were compared with radioisotopic immunoassay results and showed a good correlation coefficient, r= 0.99; Y = 0.98× + 9.3; N = 79. Sera from healthy adults has a myoglobin concentration in the range of 15‐80 ng/ml(N=362). Sex‐and age‐related differences were observed. The serum myoglobin levels in males and elderly people showed higher concentration than in females and younger people. The peak elevation of serum myoglobin compared with other cardiac markers was observed within 6 hours after onset of chest pain as well as the CK‐isoform ratio (MM3/MM1). All of the serum from 21 patients with definite acute myocardial infarction showed increased serum myoglobin levels (100‐1200 ng/ml) upon admission and within 6 hours.The results suggest that assays for serum myoglobin levels are helpful in the early diagnosis of acute myocardi

ISSN:0887-8013    

DOI:10.1002/jcla.1860060103

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractFour anti‐IgG2 monoclonal antibodies (Mabs) were evaluated for their reactivity with purified myeloma IgG2 of different light chain types and Gm allotypes in three distinct immunoenzymatic assays (ELISA). The reactivity of three Mabs with solid‐phase antigens was similar whereas an anti‐Fab antibody (clone HP 6114) predominantly bound IgG2k. In competitive and immunometric (sandwich type) assays, the binding of the two anti‐IgG2 Mabs (HP 6014 and HP 6114) reacting with epitopes located on the Fab fragment was strongly influenced by the light chain type of IgG2 and by other factors (probably including differences in the variable regions): the Mab HP 6114 reacted virtually only with IgG2k whereas the Mab HP 6014 displayed a much stronger affinity for IgG2λ than for IgG2k; for both anti‐Fab Mabs, important differences were found in their binding to individual IgG2 proteins. In addition, the Mab HP 6014 seemed to show a slightly better affinity for IgG2 bearing the Gm(23) allotype. These results urge much caution in IgG2 level measurement, especially with commerical kits, most of which use the Mab 6014 as the single anti‐

ISSN:0887-8013    

DOI:10.1002/jcla.1860060104

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractPeripheral blood samples from healthy adults, patients with transplanted bone marrow, and healthy children were analyzed for the proportions of cells positive for CD2, CD4, CD8, CD19, CD56, CDw65, and KiM8 cell membrane markers. Six cell gates were defined on granularity vs. cell size (L90°LSx‐FALS) flow cytometric display for each blood sample: large lymhocytic, small lymphocytic, monocytic mononuclear, granulocytic, and all‐cell gate, and the listed markers were determined in each of them. Considerable differences in marker‐positive cell proportions were found between healthy adults and children, and particularly between healthy adults and patients with transplanted bone marrow. Marker‐positive cell frequencies corresponded to biologic distribution of three leukocyte populations within the defined gates, in a general agreement with known specificities of the antibodies used. A considerable degree fo variations in the positions of gates drawn for different samples and the numbers of cells counted in them was observed. Still it appeared that all six gates, otherwise commonly used in flow cytometric analysis, could be precisely defined and yielded reproducible data. Large and small lymphocytic gates yielded very similar marker frequencies, revealing that in cases where three leukocyte populations were not clearly delineated on (90°LSxFALS) display, a smaller lymphocytic gate could be safely drawn in order to avoid contamination with

ISSN:0887-8013    

DOI:10.1002/jcla.1860060105

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractPolyfunctional hydrophilic microspheres of 125‐nm diameter can be produced by copolymerization of acrylic monomers. Purified c‐reactive protein (CRP) was covalently bound to these new micropheres, and the conjugate obtained was used as reagent in a microparticle‐enhanced nephelometric immunoassay for human CRP. This assay was based on the measure, with a specially designed nephelometer, of the light scattered by aggregates formed during the immunoagglutination of the conjugate with anti‐CRP antiserum. Sensitive inhibition of this agglutination by free CRP (6 ng/ml) allowed CRP quantitation in highly diluted serum samples (1/500‐1/2,000), excluding any interference or sample pretreatment. The CRP assay was easy to perform (no washing or phase separation), reliable (coefficients of variation ranged from 1.3% to 9.3% for within‐run and between‐run determinations), and accurate (mean percentage of recovery: 104%; correlation coefficients with accepted analytical methods>0.97) over a large range of concentrations. The inhibition mode excluded errors in the antigen excess zone and provided total security at high c

ISSN:0887-8013    

DOI:10.1002/jcla.1860060106

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractA radioimmunoassay (RIA) for serum thymic factor (facteur thymique serique) was established. This assay could detect<0.5pg of serum thymic factor without the interference by other thymic hormones, including thymopoietin ii and thymosin α1. Both zinc‐free and zinc‐bound serum thymic factor showed a similar standard curve, indicating that this RIA could detect both forms of serum thymic factor in the same way. We observed that two procedures were required prior to the measurement in order to measure serum thymic factor content in plasmas; the first, the extraction procedure to remove substances interfering with the assay nonspecifically; the second, the storage of the sample with ethylenediamine tetraacetic acid dissodium salt to inhibit the proteases in the plasma to degrade serum thymic factor. By employing this assay system, we observed that cord blood plasmas contained significantly higher level of serum thymic factor than did adult blood pla

ISSN:0887-8013    

DOI:10.1002/jcla.1860060107

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThe production of hydrogen peroxide (H2O2) by neutrophilic polymorphonuclear leukocytes (PMN) after stimulation and the infiltration of PMN in glomeruli were determined in 20 patients with primary IgA nephropathy. The H2O2production of PMN after the stimulation was measured with a spectrophotometer using horseradish peroxidase as substrate. The results were as follows: (1) when PMN were pretreated with cytochalasine B, H2O2production after stimulation with heataggregated IgG (IgG) or serum‐treated zymosan (STZ) was significantly higher in patients with IgA nephropathy than in controls, and (2) there was an increased amount of PMN localized in glomeruli in patients with IgA nephropathy using immunofluorescence of monoclonal anti‐PMN antibody.It appeared that the increased renal infiltration of PMN which have a high potential for production of reactive oxygen species might induce the glomerular injuries in patients with IgA nephropa

ISSN:0887-8013    

DOI:10.1002/jcla.1860060108

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractLocalization of hepatitis C virus (HCV) RNA was investigated by non‐radioactive in situ hybridization in human liver specimens of chronic non‐A, non‐B (NANB) hepatitis patients who were seropositive for antibodies to HCV (anti‐HCV). For in situ hybridization, T‐T dimerized synthetic oligodeoxynucleotide probes were used and DNAs hybridized in situ were detected immunohistochemically using specific antibodies against T‐T dimer. The data demonstrates that HCV‐RNA was localized in the cytoplasm of hepatocytes in human liver biopsies obtained from the patients with chronic NANB hepatitis seropositive

ISSN:0887-8013    

DOI:10.1002/jcla.1860060109

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractDespite their potential importance, the role of phagocyte‐derived chloramines (“longlived oxidants”) has not yet been investigated in inflammatory or infectious diseases. We have developed a sensitive spectrophotometric microtiter plate assay for chloramines based on their capacity to oxidize potassium iodide (Kl). Consistant levels of endogenous chloramines were detected in normal human polymorphonuclear neutrophil (PMN) supernatants after stimulation by phorbol myristate acetate (PMA) or opsonized zymosan. Exogenous taurine strongly enhanced chloramine secretion and was used to quantify the chlorinating potential of PMN. Taurine‐chloramines were also detectable in monocyte supenatants, although in smaller amounts. The specificity of the KI assay was assessed both in terms of effect of compounds inhibiting (KBr) or interacting with (sodium azide and catalase) chloramine formation and by showing that PMN from patients with chronic granulomatous disease, due to a hereditary lack of oxidative response capacity, were unable to produce chloramines. Taurine‐chloramine levels secreted by PMA (but not zymosan)‐stimulated PMN were closely related to the cellular luminol‐amplified chemiluminescence (CL) responses although the CL assay failed to detect chloramines in PMN supernatants. We consider that this KI assay should be of use in studying the role of long‐lived phagocyte‐derived oxidants in

ISSN:0887-8013    

DOI:10.1002/jcla.1860060110

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThis study presents data from the crossreactivity analysis of purified ciguatoxin (CTX), okadaic acid (OA), and the East sphere or Fragment B‐C of OA with their homologous antibodies, monoclonal antibodies to ciguatoxin (MAb‐CTX) and okadaic acid (MAb‐OA). The test system used was the stick enzyme immunoassay. MAb‐CTX gave peak titers of 1.5ng, 10ng and 50ng respectively for CTX, East sphere and OA. Competitive inhibition analysis showed that 4ng purified CTX blocked completely MAb‐CTX reaction with crude CTX, OA and East sphere of OA blocked at similar concentrations (approximately 50ng). The activity with MAb‐OA in the homologous system with OA and East sphere was insignificant. This may be attributable to the improper concentrations used. The cross‐reactivity between MAb‐CTX with OA and its Fragment B‐C may cause difficulty in the test system in its application to assess toxic fish

ISSN:0887-8013    

DOI:10.1002/jcla.1860060111

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY