摘要:

AbstractThe cellular nuclear antigen SS‐B/La is known to be a major antigenic target to an autoantibody in patients with Sjogren's syndrome and systemic lupus erythematosus. It is useful to detect an anti‐SS‐B/La antibody from patients' sera in a clinical point of view. We purified SS‐B/La from rabbit thymus acetone powder by affinity chromatography with a murine anti‐SS‐B/La monoclonal antibody (1C3‐H7). An enzyme‐linked immunosorbent assay method, in which SS‐B/La was used to coat a plate, was also successfully established. It is difficult to obtain a large volume of patients' serum with high antibody titer and high specificity as a positive control. We investigated whether or not a positive control from human could be replaced by a murine monoclonal antibody to SS‐B/La. The 1C3‐H7 was conjugated with a human IgG Fc' fragment using N‐γ‐maleimidobutyryloxysuccinimide as a cross‐linker. The chemically humanized murine monoclonal antibody (1C3‐Fc') was recognized by antiserum specific for human IgG Fc fragment. 1C3‐Fc' reacted to SS‐B/La but not to other antigens. Furthermore, the titration curve of this conjugate ran parallel with those of patients' sera specific for SS‐B/La. It is concluded that a chemically humanized murine monoclonal antibody is useful as a positive control in place of a human pat

ISSN:0887-8013    

DOI:10.1002/jcla.1860060602

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

年代:1992

数据来源: WILEY

摘要:

AbstractThe gene expression of human type 1 procollagen was investigated in cirrhotic human liver by using in situ hybridization with nonradioactive DNA probes. Using in situ hybridizaiton can provide direct evidence for the cell type capable for type I collagen synthesis in tissues. T‐T dimerized DNA probes were used and DNAs hybridized in situ were detected immunohistochemically using specific antibodies against T‐T dimer. The data demonstrated that type I collagen is synthesized in hepatocyles and stellate cells in pseudolobules and in fibroblasts in Glissons capsules in cirrhotic human livers. we indicated hepatocytes morphologically and functionally by using immunohistochemical localization of albumin, which ws used as a marker of hepatocyte, since albumin is synthesized exclusively by hepatocytes. © 1992 Wiley‐Lis

ISSN:0887-8013    

DOI:10.1002/jcla.1860060603

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

年代:1992

数据来源: WILEY

摘要:

AbstractThis study compared the use of conventional urine microscopy, the KOVA system, Neubauer hemacytometry, and the Yellow IRIS for the determination of white blood cell (WBC) and red blood cell (RBC) counts in urine samples. Both KOVA WBC and RBC counts correlated better with the IRIS counts than with conventional microscopy. KOVA WBC counts correlated with Neubauer hemacytometry to the same degree as they did with IRIS WBC counts. The RBC count correlation was fairly similar between KOVA, hemacytometry, and the conventional method. It was concluded that the KOVA system is a suitable replacement for conventional urine microscopy. © 1992 Wiley‐Liss, I

ISSN:0887-8013    

DOI:10.1002/jcla.1860060604

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

年代:1992

数据来源: WILEY

摘要:

AbstractWe designed a rapid, homogeneous assay for human asparatate aminotransferase (AST) isoenzymes, by a selective proteolysis of soluble AST (s‐AST), using chymotryspin and protease 401. The linearity of mitochondrial (m‐AST) was elongated up to 4000 U/l. m‐AST values from the human liver, and determined by a homogeneous assay using protease 401 or chymotryspin, were relative to those obtained using an immunoprecipitaiton method.In perioperative patients or those with an acute myocardial infarction, the peaks of s‐AST and m‐AST values were noted 13 h and at 57 h after ictus, respectively, whereas the peak of ratio between was seen 6 h after ictus. In the case of Budd‐Chiari syndrome, the maximum levels of the two AST activities were evident 14 days after hospitalization and the peak of ratio between them was seen after 7 days. We propose that htis homogeneous assay can serve as a diagnostic tool for early phase detection of myocardial infarction and of Budd‐Chiari syndrome. © 1992 W

ISSN:0887-8013    

DOI:10.1002/jcla.1860060605

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

年代:1992

数据来源: WILEY

摘要:

AbstractFive immunochemical assays for determining low concentrations of albumin were investigated. These were a radioimmunoassay (RIA); turbidimetric immunoassays (TIA) both according to end‐point measuring principle on a Cobas Fara and Hitachi 717 analysers, and according to kinetic measuring principle on a Turbitimer instrument; and a nephelometric immunoassay (NIA). All achieved the analytical goal necessary for optimal patient care. The correlations between the albumin concentrations measured with the different techniques were very good. In vitro glycation of albumin did not influence albumin concentrations measured by the five assays. Urine albumin excretion measured over 3 consecutie days showed considerable day‐to‐day variation. This was highest for spot‐urine specimens and significantly lower for 24 h and timed‐overnight samples. Variation of storage temperature (room temperature, 4°C, −20°C), time (up till 3 months), and pH (within the range pH 5–8) of the urine samples did not change significantly the measured albumin concentrations. Different sample preparations (vortex‐mixing, centrifugation, and thawing) had no influence on the measured albumin concentration. In conclusion, a maximum standardization of the collection of timed‐overnight urine samples for screening and 24 h urine sampels for confirmation of microalbuminuria during 3 consecutive days is more crucial than the choice of the immunological technique. ©

ISSN:0887-8013    

DOI:10.1002/jcla.1860060606

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

年代:1992

数据来源: WILEY

摘要:

AbstractIn order to study the concentration of pancreatic stone protein (PSP) in human pancreatic juice, we investigated the influence of the insoluble form of PSP‐S1converted from PSP‐S2–5on PSP determination and the assay method for PSP‐S1precipitate after solubilizing PSP‐S1was converted from PSP‐S2–5and precipitated about 45–85% after 1 h. The precipitated PSP‐S1was dissolved in 0.1 M sodium acetate buffer, pH4.0, and the concentration was measured by the enzyme immunoassay, with similar reactivity to PSP‐S1and PSP‐S2–5. The proposed method can offer accurate and specific analysis of the PSP level in pancreatic juice. The results of the fractionation of pancreatic juice and duodenal juice on Mono S cation‐exchange chromatograhy suggested that the major coponent of PSP was PSP‐S2–5in pancreatic juice and PSP‐S1in duo‐de

ISSN:0887-8013    

DOI:10.1002/jcla.1860060607

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

年代:1992

数据来源: WILEY

摘要:

AbstractAn automated microparticle enzyme immunoassay (MEIA) with the IMx ® analyzer for the detection of neural thread protein (NTP) in cerebrospinal fluid (CSF) from Alzheimer's disease (AD) patients was developed. This assay uses monoclonal antibodies produced against the purified pancreatic from of the protein. The assay employs one monoclonal antibody covalently coupled to the microparticle to capture immunoreactive material in CSF or brain tissue. The second monoclonal antibody was conjugated to alkaline phosphatase and serves as detection antibody. The assay provides results in approximately 45 minutes with a sensitivity of 60 pg/ml (3 fmoles/ml). The titration curve of both normal and AD CSF resulted in a linear relationship with respect to the volume of CSF used. A similar relationship was observed when normal and AD brain tissue extracts were serially diluted. The molecular weight of NTP in CSF was approximately 20 kD as determined by gel filtration method under non‐denaturing conditions. The recovery for pancreatic threat protein (PTP) spiked in either normal or AD CSF was 104% and 108%, respectively. Intra‐, inter‐, and total assay CVs (coefficient of variation) for controls were less than 2.9%, 3.3% and 3.0%, respectively. This assay will provide a useful tool in the study of the Alzheimer's disease and may help research in diagnosis and prognosis of Alzheimer's disease and related disorders. © 1992 Wiley‐

ISSN:0887-8013    

DOI:10.1002/jcla.1860060608

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

年代:1992

数据来源: WILEY

摘要:

AbstractWe report our experience with a provocative test of calcitonin (CT) release using a combined stimulus of intravenous 10% CaCl2solution and pentagastrin on 34 normal adults (15 females: age 41 ± 12.3 yers and range 22–65 years; and 19 males: age 43 ± 9.1 years and range 23–60 years) and in 44 family members of three proven multiple endocrine neoplasia type 2A syndrome (MEN 2A) patients. A commercial radioimmunoassay was used to determine the serum CT levels. Peak CT levels were reached within 2 to 5 minutes after administration of the stimulus in all subjects tested. In the group of normal subjects there was no significant difference in the mean basal CT levels between males (54.8 ± 21.7 pg/ml) and females (56.5 ± 34.8 pg/ml), whilst the mean peak response values ofr males was 146.3 ± 120.6 pg/ml, which was significantly different from the mean value of females, namely 71.6 ± 39.0 pg/ml. We did not find significant correlations between the basal CT level, peak CT response, and age.Of the 44 family members tested, 9 showed an exaggerated CT response to the combined stimulus and subsequently had a total thyroidectomy. Histological examination confirmed C‐cell hyperplasia (CCH) in one and medullary thyroid carcinoma (MTC) in teh other 8. Three of the 9 had high basal plasma CT levels. The 9 patients were retested postoperatively and all showed a flat response to the combined stimulus. Those family members with histological proof of MTC or CCH were screened for genetic linkage to the disease gene for MEN 2A using probe MCK2, and showed correltion in each instance. Eight other members were identified as high‐risk genotypes. They are still asymptomatic and calcitonin negative.Our study shows that the provocative test and the commercial calcitonin radioimmunoassay used is reliable in detecting family members of MEN type 2A patients having preclinical MTC or CCH. We found that peak response values higher than 250 pg/ml were highly suggestive of preclinical MTC in family members of MEN 2A cases (sensitivity 100% and specificity 94%). The test had minor side effects which did not las more than 2 minutes. © 1992 W

ISSN:0887-8013    

DOI:10.1002/jcla.1860060609

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

年代:1992

数据来源: WILEY

摘要:

AbstractIn this paper an uncomplicated method for the simultaneous detection and semiquantitation of 11 of the 12 commonly studied antinuclear antibodies (ANA) in a single run is described. This new application of checker‐board immunoblotting (CBIB) is based upon available technology and employs purified antigens which can be either purchased or produced in‐house. CBIB requires no electronic instrument, can be formatted to meet the needs of the user, is rapidly performed, and has acceptable labor and materials costs. Data on the use of the method to examine available reference antisera is presented. CBIB has also proven practical forthe clinical study of 18 sera, at two dilutions per membrane, for each set of specific antinuclear antibodies, also at two or more dilutions. © 1992 Wiley‐Lis

ISSN:0887-8013    

DOI:10.1002/jcla.1860060610

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

年代:1992

数据来源: WILEY

摘要:

AbstractThe intercalibration precision and linearity were determined for two representative analytes, apolipoprotein A1 (apo A1) and immunoglobulin G (IgG), on the Beckman Array® and the Behring Nephelometer 100® (BN‐100). For two of nine samples analyzed, poor precision was observed for IgG with the Array. The poor precision for these two samples is attributed to large systematic shifts. A statistical non‐linearity was observed for apo A1 with the BN‐100, but this non‐linearity does not exclude use of this assay for clinical diagnosis. Method comparisons were done for 12 analytes: apo A1, apo B, IgG, IgA, IgM, IgE, C3, C4, albumin, C‐reactive protein, alpha‐1‐antitrypsin, and ceruloplasmin. Thjese comparisons showed proportional biases of>10% for seven of 12 analytes. Furthermore, correlation coefficients were<0.96 for seven of 12 analytes. We conclude that comparing results obtained from th two different nephelometers is of only limited value for the individual patient. © 1992

ISSN:0887-8013    

DOI:10.1002/jcla.1860060611

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

年代:1992

数据来源: WILEY