摘要:

AbstractSm antibodies are specific serum markers for the diagnosis of systemic lupus erythematosus (SLE) and are identified by immunodiffusion (ID), counterimmunoelectrophoresis (CIE), and passive hemagglutination (PHA) in the clinical laboratories. These current methods have certain disadvantages of sensitivity, specificity, and complexity.In this report, Sm antigen free from other nuclear antigens was purified. Synthetic particles were coated with Sm antigen and used to test for Sm antibodies in patient's sera. The particle agglutination test was specific for only Sm antibodies among different antinuclear antisera from patients with systemic rheumatic diseases. The particle agglutination test had the same sensitivity and specificity as the enzyme‐linked immunoassay (ELISA) test for Sm and RNP antibodies. The correlation coefficient between the ELISA quantitation and the titers obtained by particle agglutination was 0.82. The particle agglutination test can be used for rapid screening and titer determination of anti‐Sm antibod

ISSN:0887-8013    

DOI:10.1002/jcla.1860010402

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

年代:1987

数据来源: WILEY

摘要:

AbstractA diagnostic test for intestinal amebiasis was developed by using an ELISA that incorporated a well‐defined monoclonal antibody specific for a membrane antigen ofEntamoeba histolytica. The ELISA was found to be sensitive and quite specific forE. histolytica, since there were no cross reactions with other parasites, especiallyEntamoeba coli, with whichEntamoeba histolyticais often confused. In addition, this test detected not onlyE. histolyticatrophozoites but also cysts. This technique will be of great value in the rapid and accurate diagnosis ofE. histolyticain human fecal materia

ISSN:0887-8013    

DOI:10.1002/jcla.1860010403

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

年代:1987

数据来源: WILEY

摘要:

AbstractAn immunoassay using fluorescent microspheres was developed for the detection of cell surface antigens on biopsied human cancer cells. A murine IgM monoclonal antibody 202 that was raised against a human melanoma cell line detected monosialogangliosides having sialic acid‐galactose residue, and was used as an antibody source. Purified 202 antibody was coupled with 1‐μm fluorescent micro‐spheres (Immunosphere) by carbondiimide reaction at pH 4.7–5.5, and the assay was standardized using an antigen‐positive human melanoma cell line. Over 90% of cells were positive when 4 μg antibody‐containing immunospheres were used. The specificity of the reactivity was demonstrated by a blocking assay using uncoupled 202 antibody. The binding activity was stable for 2 weeks after preparation.Biopsied specimens that had been viably frozen in a form of single‐cell suspension were thawed and purified by Ficoll density gradient centrifugation. All of the five melanoma specimens tested were positive (>5% fluorescent cells) with a mean percent of positive cells = 35%, whereas only two of the five nonmelanoma tumor specimens were positive (m = 10.8%). None of the ten noncancer tissues, including skin, liver, kidney, and lymphocyte cells, showed greater than 5% positivity. When these tissues were tested after a 3‐day incubation of the cells in a CO2incubator, a significant increase in the antibody reactivity was obtained in melanoma tissues (m = 57.8%). In contrast, the antigenic expression of other types of human cancer was changed only slightly (m = 16.2%) within the same period of incubation. Whether the assay is applicable to cell surface antigens of other epitopes of human biopsied tissues or only to ganglioside antigens is currently un

ISSN:0887-8013    

DOI:10.1002/jcla.1860010404

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

年代:1987

数据来源: WILEY

摘要:

AbstractA sensitive and reproducible method for quantitation of polyamines in erythrocytes by high‐performance liquid chromatography (HPLC) is described. Spermidine and spermine are first converted to fluorescent dansyl derivatives and are then separated in less than 15 minutes on a C18 reverse‐phase cartridge using methanol in water mobile phase. Sensitivity of the method is 20 pmol with recoveries that average 96% and 95% for spermidine and spermine, respectively. Precision study revealed an average mean coefficient of variation of 5.42 ± 2.95 (mean ± S.D.) from four levels of the polyamines (8.2 to 354.9 nmol/g hemoglobin [Hgb]). The normal ranges for 30 apparently healthy men and women, aged 20–60 years, were 2.9–33.7 nmol/g Hgb (± 2 S.D. from mean) for spermidine and 0‐20.9 nmol/g Hgb for spermine. The quantity of putrescine was negligible. Feasibility of this method was evaluated in serial specimens from an advanced‐stage patient with prostate cancer who was receiving multiple‐modality therapy. Results revealed that this HPLC method can be used in quantifying circulating polyamines in cl

ISSN:0887-8013    

DOI:10.1002/jcla.1860010405

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

年代:1987

数据来源: WILEY

摘要:

AbstractChemosensitivity testing has usually been limited to tumor specimens of 2–3 g or more. We evaluated the miniaturized, improved nucleic acid precursor incorporation assay (MINI‐assay) in 100 small biopsy specimens of 1 g or less. Tumor types included malignant melanomas; sarcomas; and breast, colon, lung, and ovarian carcinomas. Overall, 74/100 were evaluable. The evaluability rate was related to size: 41/48 (85%) 0.7–1.0‐g specimens were evaluable, as opposed to 22/35 (64%) 0.4–0.6‐g and 11/17 (64%) 0.1–0.3‐g specimens. Evaluability was not correlated with tumor types or whether the tumor was from a primary or metastatic site, but was related to yield of viable tumor cells. No viable tumor cells were obtained in 17 specimens, and thus no assay was performed. When 105cells per well were plated, 49/54 (91%) assays were evaluable, but only 25/29 (86%) were successful when fewer cells were used. Four or more anticancer drugs could be tested in 69/74 (95%) of the evaluable assays. Yielding high evaluability rates and requiring fewer viable cells, the MINI‐assay has allowed us to perform chemosensitivity tests on previously unusable

ISSN:0887-8013    

DOI:10.1002/jcla.1860010406

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

年代:1987

数据来源: WILEY

摘要:

AbstractLiver‐kidney‐microsomal (LKM) autoantibodies are diagnostic markers for a subgroup of HBsAg‐negative chronic active hepatitis, presumably owing to autoimmunity. They were originally detected by indirect immunofluorescence and can now be evaluated by radioimmunoassay, enzyme‐linked immunosorbent assay, and immunoblotting. In immunoblotting LKM‐positive sera react strongly with a 50‐kilodalton (KD) polypeptide band of microsomes. In immunoelectron microscopy, LKM‐positive sera show a binding with membranes of the endoplasmic reticulum. The LKM antigen was further identified on various isoenzymes of cytochrome P‐450. Immunofluorescence is still the method of choice for screening sera routinely. However, cytplasmic antigen‐antibody systems often can hardly be distinguished by this method. A specific radioimmunoassay and an enzyme‐linked immunosorbent assay can differentiate the various autoantibodies against cytoplasmic antigens. Immunoblotting and immunoelectron microscopy are specific tools for the characterization of the target antigens on an ultrastructural or molecular level. So far they have no use in routine testing of sera. However, since LKM antigen was localized on isozymes of cytochrome P‐450, this subgroup of CAH might turn out to be a drug‐induced autoimmune liver disease. Clinically, these patients are characterized by chronic active hepatitis or cirrhosis in liver histology, a slight predominance of females, low IgA immunoglobulin levels, and an often rapidly progressing disease. Patients can be treated with immunosuppressive drugs. However, controlled therapeutical trials are missing. Furthermore, an immunogenetic background still has to be proven for this a

ISSN:0887-8013    

DOI:10.1002/jcla.1860010407

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

年代:1987

数据来源: WILEY

ISSN:0887-8013    

DOI:10.1002/jcla.1860010408

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

年代:1987

数据来源: WILEY

摘要:

AbstractChronic inflammatory liver diseases are classified as chronic active hepatitis (CAH) or primary biliary cirrhosis (PBC) on the basis of clinical and pathological criteria. Autoantibodies allow the distinction between autoimmune forms of CAH and virally induced forms, in particular CAH non‐A, non‐B. Three different subgroups of autoimmune‐type CAH have been defined so far. Classical autoimmune “lupoid” CAH is characterized by antinuclear antibodies (ANA) and liver‐membrane autoantibodies (LMA). Antibodies against a liver‐kidney‐microsomal antigen (LKM) are found in a small group of generally young patients and are associated with a progressive form of autoimmune CAH. The target antigen as defined by immunoblotting is a protein at 50 kilodaltons (KD), and it has been detected on phenobarbital‐inducible isoenzymes of cytochrome P‐450. The third subgroup, clinically similar to “lupoid” CAH, is characterized by antibodies against a soluble cytoplasmic liver antigen (anti‐SLA). These antibodies cannot be diagnosed by routine methods, i.e., immunofluorescence, but are only found by radioimmunoassay or enzyme immunoassay. In primary biliary cirrhosis (PBC) the immune reactions are directed against the intrahepatic bile ducts rather than hepatocytes. Antimitochondrial antibodies (AMA), traditionally diagnosed by immunofluorescence, are found in up to 100% of PBC patients. Complement‐fixation test, radioimmunoassay, and enzyme immunoassay allow the distinction of disease‐specific subtypes of AMA. Immunoblotting allows the characterization at the molecular level of two PBC‐specific AMA subtypes, anti‐p48 and anti‐p62. The latter corresponds to the anti‐M2 antibodies described previously. All these antibodies are important tools in the differential diagnosis of chronic inflammatory liver diseases. Patients with autoimmune‐type CAH benefit from immunosuppressive therapy, while those with virally induced CAH do not. The role of these autoantibodies in the pathogenesis of CAH or PBC is not known as yet, but they are se

ISSN:0887-8013    

DOI:10.1002/jcla.1860010409

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

年代:1987

数据来源: WILEY

摘要:

AbstractThe porphyrias, a group of diseases caused by enzyme deficiencies in the heme biosynthetic pathway, are reviewed. The clinical features and pathophysiology of each porphyria are discussed. Analytical techniques used in both screening and definitive investigations are reviewed. The final sections integrate the previous information to provide a protocol to assist in planning the investigation of any particular case of porphyria according to clinical presentation and in interpreting the results of laboratory investigations.

ISSN:0887-8013    

DOI:10.1002/jcla.1860010410

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

年代:1987

数据来源: WILEY

ISSN:0887-8013    

DOI:10.1002/jcla.1860010401

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

年代:1987

数据来源: WILEY