bluuebunny/crossref_metadata_2025_split huggingface dataset:
Methodology and significance of the detection of liver‐kidney‐microsomal (lkm) autoantibodies in autoimmune‐type chronic active hepatitis
Authors: Manns M. , Gerken G. , Kyriatsoulis A. , zum Büschenfelde K.‐H. Meyer , Dienes H. P. 
Published: March 2007

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 autoimmune liver disease.
Significant autoimmune markers of autoimmune liver disorders: Current status
Authors: Manns M. , Gerken G. , Kyriatsoulis A. , zum Büschenfelde K.‐H. Meyer 
Published: March 2007

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 sensitive and specific diagnostic markers.
Automated nephelometric immunoassays with novel shell/core particles
Authors: Kapmeyer Wolfgang H. , Pauly Hans‐Erwin , Tuengler Peter 
Published: March 2007

AbstractNephelometry has become a very reliable and convenient method for immunoassays. Recent innovations in polymer chemistry have led to the design of highly sensitive and reproducible immunoassays for the Behring Nephelometer Analyzer. The principle of particle‐enhanced immunoassays has led to the development of new polymer particles. Key advances include the preparation and use of particles with polystyrene cores covered by thin shells with chemically reactive groups. The cores are prepared by emulsion polymerization, giving particles with a well‐defined size. The particle size has been optimized to fulfill the requirements for nephelometric analysis. The shells consist of a polystyrene/polymethacrylate/polymethacrylamidoacet‐aldehyde dipentylacetal copolymer, which allows a covalent binding of the immunochemicals of interest. Using these particles, a method for the determination of C‐reactive protein has been worked out.
Direct measurement of α‐human atrial natriuretic polypeptide in plasma by sensitive enzyme immunoassay
Authors: Hashida Seiichi , Ishikawa Eiji , Mukoyama Masashi , Nakao Kazuwa , Imura Hiroo 
Published: March 2007

Abstractα‐Human atrial natriuretic polypeptide (α‐hANP) in plasma was directly measured without extraction by sensitive sandwich enzyme immunoassay. Polystyrene balls were coated with monoclonal anti‐α‐hANP (ring) IgG, specific for the N‐terminal half of the ring structure including Met‐12 residue of α‐hANP or monoclonal anti‐α‐hANP (N‐terminus) IgG, specific for the N‐terminus of α‐hANP. Rabbit anti‐α‐hANP (C‐ter‐minus) Fab' specific for the C‐terminus of α‐hANP was conjugated to horseradish peroxidase. The polystyrene ball was incubated with α‐hANP standards or plasma and, after washing, with the conjugate, and bound peroxidase activity was assayed by fluorimetry (two‐step sandwich enzyme immunoassay). The detection limit of α‐hANP was 30–90 fg (10–30 attomo1es)/tube and 0.6–2.3 ng (0.2–0.75 pmol)/L using 0.04–0.05 ml of plasma. The lower detection limit was obtained using monoclonal anti‐α‐hANP (ring) IgG1. Plasma hANP levels of healthy men in a supine position after an overnight fast were 24.5 ± 13.2 (SD) ng/L and tended to decrease (15.3 ± 8.5 [SD] ng/L) after intravenous administration of furosemide and subsequent one hour walking. The present sandwich enzyme immunoassay could be modified further to improve the detection limit of plasma 01‐hANP (0.2 ng/L) or to perform a less time‐consuming one‐step sandwich enzyme immunoassay without much loss of the sensitivity.
Relationship between the target antigen of liver‐kidney microsomal (LKM) autoantibodies and rat isoenzymes of cytochrome P‐450
Authors: Manns Michael , Kyriatsoulis Apostolos , Gerken Guido , Lohse Ansgar , Zum Büschenfelde Karl‐Hermann Meyer , Amelizad Zahra , Oesch Franz , Reske Konrad 
Published: March 2007

AbstractChronic active hepatitis (CAH) is a clinical syndrome of different etiologies. Liver‐kidney microsomal (LKM) autoantibodies characterize a subgroup of HBsAg negative CAH, which is considered to be an autoimmune liver disease. By immunoblotting analysis (IB) LKM positive sera have been shown to react strongly with a poly‐peptide band at 50 kD. Therefore we investigated various rat microsomal enzymes with a molecular weight around 50 kD as potential candidate target antigens. These included epoxide hydrolase, cytochrome P‐450 reductase, and phenobarbital‐inducible isoenzymes of cytochrome P‐450 (PB1, PB2, PB3a, PB3b). By radioimmunoassay (RIA) and IB LKM positive sera were shown to react with the phenobarbital‐inducible isoenzymes of cytochrome P‐450 but not with epoxide hydrolase or with cytochrome P‐450 reductase. Cytochrome P‐450 PB3a and PB3b were both able to absorb exhaustively the reactivity of LKM sera with liver tissue in immunofluorescence (IF) and with purified rat liver microsomes in RIA. It is concluded that LKM antibodies are directed against an epitope present in preparations of various isoenzymes of cytochrome P‐450.
Flow cytometry analysis of platelet antibodies
Authors: Finley Paul R. , Williams R. Jane , Fletcher Carla 
Published: March 2007

AbstractWe have devised assays to detect both circulating alloantibodies to platelets (indirect assay) and platelet‐association IgG and IgM (direct assay) using a flow cytometric technique. A variety of patients with immune thrombocytopenia were studied. Employment of a confocal lens in the flow cytometer increased the discrimination power of the instrument. Patients with autoimmune thrombocytopenia (idiopathic thrombocytic purpura [ITP], systemic lupus erythematosus (SLE), lymphoma, leukemia, and drug‐induced thrombocytopenia showed a significant increase in platelet‐associated antibody. Circulating antibodies to platelets (alloantibodies) were demonstrated in cases of platelet refractoriness and neonatal isoimmune purpura. Day‐today precision of the assays ranged from 3% to 6% (coefficient of variation). No interference was shown in the presence of hemoglobin (5 g/L), triglycerides (10 g/L), or polyclonal and monoclonal immunoglobulinemia (50 g/L: IgG, IgA, IgM). The sensitivity of the direct assay was 500 attograms of IgG or IgM platelet.
Flow cytometric analysis of lymphocyte subpopulations in infants with congenital heart disease
Authors: Slade Herbert B. , Greenwood Jay H. , Beekman Robert H. , Hudson Jerry L. , Pahwa Savita , Schwartz Stanley A. 
Published: March 2007

AbstractPremortem diagnosis of the DiGeorge syndrome and its partial variants relies on the demonstration of a primary defect in cell‐mediated immunity, generally in the setting of an infant with congenital heart disease, hypocalcemia, absence of a thymic shadow, and typical dysmorphic features. Although T‐cell enumeration is considered a vital part of the diagnostic evaluation, no studies to date have addressed the issue of appropriate reference data in infants with congenital heart disease. We therefore undertook a prospective descriptive study of lymphocyte phenotype analysis in 27 nontransfused infants undergoing diagnostic cardiac catheterization. Striking differences were seen between patients and adult controls in means percentages and numbers of most lymphocyte subsets analyzed. Few differences were found in comparing the patient data to values for age‐matched control infants without heart disease. The data are discussed with reference to published values for patients with partial DiGeorge syndrome. It is concluded that lymphocyte phenotype analysis in the diagnostic evaluation of patients with suspected DiGeorge syndrome must utilize appropriate reference values.
Detection of anionic sites and immunoglobulin a deposits in the glomerular capillary walls from patients with Iga nephropathy
Authors: Tomino Yasuhiko , Yagame Mitsunori , Eguchi Kazuhiko , Miyazaki Masanobu , Nomoto Yasuo , Sakai Hideto , Shirato Isao , Ito Kiichi 
Published: March 2007

AbstractA study of anionic sites in the glomerular basement membrane (GBM) from patients with immunoglobulin A (IgA) nephropathy is described. The relationship between the deposition of IgA and the detection of glomerular extracellular components, i.e., noncollagenous (NC‐1) domain of Type IV collagen, in the glomerular capillary walls was examined by double immunofluorescence. Renal biopsy specimens from patients with IgA nephropathy were immersed in polyethyleneimine (PEI) as a cationic probe and then examined by electron microscopy. Renal specimens were also incubated with mouse monoclonal anti‐NC‐1 domain of Type IV collagen and then stained with fluorescein isothiocyanate (FITC)‐labelled goat antimouse lg antiserum. After these reactions, sections were stained with rhodamine‐labelled rabbit antihuman IgA antiserum. GBM subepithelial anionic sites marked by PEI were altered by the deposition of electron‐dense deposits (EDD) in patients with IgA nephropathy and there was a significant correlation between the levels of proteinuria and the incidence of EDD in the GBM in such patients. Marked proteinuria was observed in patients who showed loss of anionic sites in the GBM by electron microscopy. By double immunofluorescence, IgA was shown to be focally deposited outside the NC‐1 domain of Type IV collagen‐detected regions in the same patients. The authors concluded that high levels of proteinuria might be due to alterations of the size barrier and/or anionic sites of GBM in the moderate stage of IgA nephropathy.
Cholesterol assays in the seralyzers® instrument
Authors: Lott John A. , Miller Irene L. , Crowley Lawrence E. 
Published: March 2007

AbstractWe evaluated the Seralyzer® instrument for the assay of serum cholesterol and compared it to the Kodak Ektachem™ method. The Seralyzer showed good accuracy in the analysis of cholesterol in Abell‐Kendall‐verified serum pools, and the bias from the expected value was small in all cases but one. The Seralyzer exhibited CVs of <5% in all cases and good comparison with the Ektachem method. The Seralyzer is easy to use; however, some training in the proper pipetting technique is necessary. The Seralyzer meets medical needs criteria of total error >5% from the true cholesterol value.
Serum levels of cancer‐associated antigen ca‐195 in gastrointestinal cancers and its comparison with ca19‐9
Authors: Bhargava Arvind K. , Petrelli Nicholas J. , Karna A. , Parshall Philip L. , Fitzpatrick John E. , Douglass Harold O. , Herrera Lemuel , Bray Kurt , Gaur Pramod 
Published: March 2007

AbstractThe antibody against CA‐195 binds to an epitope that consists of both Lewis A and sialylated Lewis A blood group antigen, whereas CA19‐9 has shown specificity for sialylated Lewis A blood group antigen. CA19‐9 and CA‐195 levels were measured in the sera of 52 normal subjects; 65 benign disease patients; and 74 non‐gastrointestinal, 149 colorectal, and 119 upper gastrointestinal cancer patients to correlate their levels with disease status of the patients. Low incidence and levels were found among normal subjects for both markers; however, in the benign disease group a slightly higher incidence of elevation was seen for CA19‐9. Among colorectal cancer patients CA‐195 appeared to show higher sensitivity for primary as well as advanced disease. Levels of both markers showed similar incidences of elevation among upper gastrointestinal cancer patients. Based on these results the contribution of Lea specificity of CA‐195 cannot be ruled out, and it may be used alone or in combination with other markers for monitoring of patients with colorectal, pancreatic, gastric, gall bladder, bile duct, and liver cancers.
T lymphocytes possess receptors for brain myelin small protein
Authors: Riccio P. , Colangelo A. M. , Jirillo E. , Munno I. , Bobba A. 
Published: March 2007

AbstractIn immunomediated demyelinating diseases, T cells are found in chronic lesions. To discover whether immunocompetent cells may interact with some myelin proteins, we purified myelin proteins in the lipid‐bound native state and evaluated their binding to peripheral blood mononuclear cells (PMBC) isolated from healthy donors. To this end, myelin proteins were conjugated to biotin and added to PBMCs or purified CD4+ and CD8+ cells; then binding was detected using fluoresceinated avidin. In this article, we describe experiments carried out with a myelin protein recently discovered in the central nervous system. Our results show that this small, phosphatidylserine‐binding protein can bind to human T cells.
Effects of radiolabelled monoclonal antibody infusion on blood leukocytes in cancer patients
Authors: Gridley Daila S. , Slater James M. , Stickney Dwight R. 
Published: March 2007

AbstractThis study was undertaken to investigate the effects of a single infusion of radiolabelled murine monoclonal antibody (MAb) on peripheral blood leukocytes in cancer patients. Eleven patients with disseminated colon cancer, malignant melanoma, or lung adenocarcinoma were infused with 111In‐labelled anti‐ZCE 025, anti‐p97 type 96 9, or LA 20207 MAb, respectively. Blood samples were obtained before infusion, immediately after infusion (1 hr), and at 4 and 7 days postinfusion. Flow cytometry analysis of CD3+, CD4+, CD8+, CD16+, and CD19+ lymphocytes showed increasing CD4:CD8 ratios in seven patients after infusion. This phenomenon was not restricted to antibody subclass or to type of cancer. Two of the remaining patients exhibited a marked post‐infusion increase in CD8+ cells. In all three patients with malignant melanoma, decreasing levels of CD16' lymphocytes were noted after infusion and natural killer cell cytotoxicity showed fluctuations which paralleled the changes in the CD16+ subpopulation. Oxygen radical production by phagocytic cells was markedly affected in three subjects. These results suggest that a single infusion of radiolabelled murine MAb may alter the balance of critical lymphocyte subpopulations and modulate other leukocyte responses in cancer patients.
A simple and rapid procedure for the direct detection of cytomegalovirus in urine samples
Authors: Landini M. P. , Guan M. X. , Ripalti A. , Lazzarotto T. , La Placa M. , Trevisani B. 
Published: March 2007

AbstractWe compared cytomegalovirus (CMV) isolation in tissue culture and viral DNA detection by DNA:DNA hybridization in 60 clinical urine samples concentrated by different procedures. Our results showed that urine concentration by filtration is the easiest and quickest procedure allowing further detection of CMV. With optimized working conditions, the CMV detection in urine can be done in 5 hr without requiring cell cultures or costly instruments.
Potential clinical applications of c‐reactive protein
Authors: Okamura J. M. , Miyagi J. M. , Terada K. , Hokama Y. 
Published: March 2007

AbstractThis article reviews the clinical applications of C‐reactive protein (CRP). This acute‐phase protein is a distinct and sensitive marker for inflammation and tissue injury. It is a simple, fast, and relatively inexpensive latex agglutination test. The aspects of CRP reviewed include diagnostic support, serial measurements to evaluate disease course and therapeutic response, and screening studies.
Lytic units reconsidered: Pitfalls in calculation and usage
Authors: Pollock Raphael E. , Fuchshuber Pascal , Lotzová Eva , Zimmerman Stuart O. 
Published: March 2007

Abstract51 Chromium release‐derived cytotoxicity data yield curvilinear plots when the x axis displays the effector: target ratio and the y axis displays the percentage of cytotoxicity. To facilitate data analysis, several biomathematical models (simple linear regression, exponential fit, and Von Krogh) have been used to express these cytotoxicity curves as a single numerical value, termed the lytic unit. Other than using raw cytotoxicity data, the lytic unit has been the most common method of data presentation in human and animal tumor immune studies involving natural killer cells, lymphokine‐activated killer cells, and cytotoxic T cells. Unfortunately, the models for determining lytic unit values incorporate assumptions and methods of calculation that can result in inaccurate model‐predicted cytotoxicity in comparison with the actual observed cytotoxicity data. Even when the model is accurate in predicting cytotoxicity values (i.e., the nonlinear regression‐calculated three‐parameter Von Kqh model), comparisons between donors of minimally different or highly different cytotoxicity are still fraught with potential error due to statistically verifiable violations of assumptions of parallelism. Although more cumbersome, donor cytotoxicity comparisons using a range of effector: target ratios are not subject to the above problems. Researchers may therefore want to reconsider the use of lytic units when evaluating and reporting cytotoxicity data.
Assay for the detection of anti‐idiotypic antibodies to monoclonal antibody b72.3
Authors: Ferroni Patrizia , Milenic Diane E. , Schlom Jeffrey , Colcher David 
Published: March 2007

AbstractThe administration of xenogenic monoclonal antibodies (MAbs) leads in many cases to a host immune response represented by the generation of antibodies that can be directed against allotypic, isotypic, and idiotypic determinants present on the xenogeneic MAb. Anti‐idiotypic antibodies (Ab2s) can be detected by measuring their specific reactivity in sandwich assays using their ability to cross‐link labeled Abl to Abl attached to a solid phase; however, when the MAb used for these studies reacts with a multideterminant tumor‐associated antigen found in the circulation (e.g., TAG‐72), the utility of these anti‐idiotype assays may be limited. To determine the levels of anti‐idiotypic antibodies that could be detected in patients undergoing MAb B72.3‐based immunodiagnostic and immunotherapeutic protocols, we investigated the ability of a solid‐phase sandwich radio‐immunoassay (RIA) to detect anti‐idiotypic antibodies raised against 872.3. Furthermore, to overcome the interference of circulating TAG‐72 and for antibodies to allotypic and isotypic determinants in the evaluation of an anti‐idiotypic response, we developed a methodology using CC49‐coated resin to adsorb the interfering molecules (CC49 is a second anti‐TAG‐72 MAb). Under the conditions established, all of the TAG‐72 antigen was removed by adsorption with MAb CC49. Furthermore, since the treatment used an isotype‐identical murine MAb, the binding due to the anti‐mouse antibodies, other than the anti‐idiotype, was completely abolished after a treatment with MAb CC49, leaving only the anti‐idiotype component. Analysis of serum samples from patients who had received 872.3 that were positive for human anti‐mouse antibodies in the 872.3 sandwich RIA, after the adsorption with CC49 resin, revealed the presence of a 872.3–binding component in 2 of 12 samples. The ability of the adsorbed sera to compete with an anti‐B72.3‐idiotype MAb for the binding sites present on the MAb 872.3 confirmed the anti‐idiotypic nature of the component being detected in the patient sera.
Bilirubin diglucuronide as the main source for in vitro formation of delta bilirubin
Authors: Adachi Yukihiko , Kambe Akira , Yamashita Masaki , Yamamoto Toshio 
Published: March 2007

AbstractTo clarify which of the bilirubin moieties is responsible for the formation of bilirubin bonded to albumin (delta bilirubin) in icteric serum, the in vitro formation of delta bilirubin from bile acid‐free bilirubin glucuronides and unconjugated bilirubin was examined by high‐performance liquid chromatography. Bovine serum albumin (150 μmol/liter) was mixed with equimolar bilirubin diglucuronide (BDG), bilirubin monoglucuronide (BMG), or unconjugated bilirubin (UCB) and incubated in the dark at 37°C under argon gas saturation. Although no delta bilirubin was formed immediately, formation eventually occurred and increased with time. A similar amount of delta bilirubin was formed when human serum albumin was used instead of bovine serum albumin. Of the three types of bilirubin, BDG was found to be the greatest source of delta bilirubin, whereas UCB produced the least. On the other hand, photoirradiation of a mixture of bovine serum albumin and UCB at a molar ratio of 1:1 resulted 6 hr later in the formation of three times as much delta bilirubin as in nonirradiated specimens. This photoinduced delta bilirubin formation increased further when the UCB/albumin molar ratio was increased to 2:1.