bluuebunny/crossref_metadata_2025_split huggingface dataset:
Serum pseudocholinesterase and very‐low‐density lipoprotein metabolism
Authors: Kutty K. M. , Payne R. H. 
Published: March 2007

AbstractSerum pseudocholinesterase (PChE) was discovered in 1932. Since this protein mimics many of the catalytic properties of acetylcholinesterase, it has traditionally been referred to as PChE, even though its true biological function is unknown. Serum PChE is synthesized in the liver and secreted into the circulation as a sialated glycoprotein. Although no convincing evidence of biological function exists, a significant number of obese and diabetic patients have elevated levels of PChE. The same phenomenon is found in experimental animal models of obesity, diabetes and hyperlipoproteinemia. Streptozotocin‐induced diabetic mice showed increased serum PChE activity concomitant with increased serum triacylglycerol and PChE activity declined with treatment. Iso‐OMPA, a nontoxic inhibitor of serum PChE, reduced serum and liver triacylglycerols and serum VLDL in streptozotocininduced rodent diabetes. These findings suggest that PChE may have a role in VLDL metabolism. © 1994 Wiley‐Liss, Inc.
Comparison of interferon‐γ and interleukin‐4 production by peripheral blood mononuclear cells and isolated T cells after activation with polyclonal t cell activators
Authors: Gonzalez Salvador , Beck Lucinda , Wilson Nevin , Spiegelberg Hans L. 
Published: March 2007

AbstractControversial data have been reported regarding the ability of peripheral blood T cells to secrete interferon‐γ (IFN‐γ) and interleukin‐4 (IL‐4) from atopic patients as compared to nonatopic healthy controls. In most of these studies, T cells in peripheral blood mononuclear cell preparations (PBMC) were stimulated with polyclonal T cell activators. Some of these activators are able to activate cells other than T cells in the PBMC preparations which may influence the lymphokine levels in supernatants of PBMC. To evaluate this, we compared the IFN‐γ and IL‐4 levels in PBMC and isolated T cell preparations after activation with phytohemagglutinin (PHA), Concanavalin A (ConA), anti‐CD3 plus phorbol myristate acetate (PMA), or ionomycin plus PMA. The IFN‐γ and IL‐4 levels in the supernatants were calculated based on the percent T cells in the preparations. Whereas all activators induced significant IFN‐γ secretion, only ionomycin plus PMA stimulation induced large IL‐4 secretion. In virtually all cases, the IFN‐γ levels calculated on a per T cell basis differed for PBMC versus isolated T cells. Whereas in some donors the IFN‐γ levels were higher in PBMC preparations than in T cells, in others it was the opposite. Similarly, in about one half of both normal and atopic donors tested, the IL‐4 levels of activated PBMC were 2‐ to 7‐fold lower than levels in isolated T cells. The data suggest that non‐T cells have a significant effect on the IFN‐γ and IL‐4 levels in supernatants of polyclonally activated PBMC. This indicates that isolated T cells rather than PBMC should be analyzed for determining possible differences in the ability of T cells from patients or normals to secrete lymphokines. © 1994 Wiley‐Liss, Inc.
Detection of autoantibodies against M2, LKM‐1, and SLA in liver diseases by standardized uniform ELISA‐techniques
Authors: Gruber R. , Felber E. , Pape G. R. , Höchtlen‐Vollmar W. , Riethmüller G. 
Published: March 2007

AbstractAntibodies directed against soluble liver antigen (SLA), liver kidney microsomal antigen (LKM‐1‐AG), and antimitochondrial antigen M2 (M2‐AMA) are critical serological markers for the differential diagnosis of autoimmune chronic active hepatitis (AI‐CAH) and primary biliary cirrhosis (PBC). The exact diagnosis of autoimmune hepatitis and PBC is of great clinical relevance, as it leads to different therapeutic strategies.In the present work, a simple and reliable ELISA test system is described, which applies the same test principle for the detection of three different species of autoantibodies important for the diagnosis of chronic liver disease. The ELISA assays are based on a competitive inhibition of binding of positive standard antibodies by patients sera containing antibodies of unknown specificity. The purified immunoglobulins of clinically and serologically clearly defined patients with SLA or LKM‐1 positive AI‐CAH and with M2 positive PBC were used as coating‐ and detection antibodies in the ELISAs. From homogenized rat liver the fractionated 100,000g supernatant was employed for the SLA ELISA, the microsomal preparation served as antigen for the LKM‐1 ELISA and the mitochondrial preparation was used for the M2 ELISA. In 1,500 sera of patients with the differential diagnosis of a hepatobiliary disease, 17 gave a positive signal in the SLA ELISA, 12 in the LKM‐1‐ELISA, and 72 in the M2‐ELISA. The results of the ELISAs were compared with Western blotting and immunofluorescence staining pattern on cryostat sections and Hep2 cells. The antibody profiles of several patients are described in detail. © 1994 Wiley‐Liss, Inc.
Successful use of two rapid HCV assays in a high prevalence romanian population
Authors: Constantine Niel T. , Holm‐Hansen Carol , Skaug Nils , Vasilescu Florica 
Published: March 2007

AbstractHepatitis C virus (HCV) infection is of significant concern for recipients of blood and for patients who share resources such as hemodialysis machines. In many developing countries and in small peripheral laboratories, facilities and capabilities to perform ELISA assays may not be available. We evaluated two newly marketed rapid HCV serologic assays to determine their ability and suitability to detect antibodies to HCV in hemodialysis patients in Bucharest, Romania. Results indicated that both the Rapid HCV Ab assay (Clonatec, Paris, France) and the HCV‐SPOT (Diagnostic Biotechnology, Singapore) detected HCV antibody in 23 of 27 patients. All 23 samples were also reactive by a routine HCV ELISA and were confirmed using a supplemental assay (HCV blot, Diagnostic Biotechnology, Singapore). Only one sample produced equivocal result by the Rapid HCV Ab assay. In this high prevalence population (90%), the use of recombinant or synthetic peptide rapid HCV assays has been successful in detecting confirmed positive cases of HCV and has shown excellent correlation with an ELISA screening assay. The tests are simple to perform, can be performed by individuals with minimal training, and have built‐in quality control measures. We conclude that these tests may have important applicability in certain testing situations. © 1994 Wiley‐Liss, Inc.
Current status of available standards for quality improvement of assays for detection of autoantibodies to nuclear and intracellular antigens
Authors: Nakamura Robert M. , Bylund David J. , Tan Eng M. 
Published: March 2007

AbstractConsiderable progress has been made in the improvement of clinical assays for the detection of autoantibodies to nuclear and intracellular antigens with the use of available World Health Organization (WHO) and Arthritis Foundation/Centers for Disease Control (AF/CDC) standards. The ultimate goal of standardization is for various clinical laboratory test results to be interchangeable and for an exchange of data to be done with confidence.This report discusses the available standards. In addition, significant technical problems and variations in methodologies for the detection of autoantibodies to intracellular antigens noted during a 4‐year study by a European Consensus Study Group are detailed.Currently, there is a need for a future generation of reference preparations and standards that will show specific antibody reactivity on sensitive enzymes and immunoblotting assays. Standardization efforts should be done to characterize specific nuclear and cellular antigen preparations that may be of natural or of recombinant technology origin. © 1994 Wiley‐Liss, Inc.
Chemiluminescent immunoassays: Discrimination between the reactivities of natural and human patient antibodies with antigens from eukaryotic pathogens, Trypanosoma cruzi and Paracoccidioides brasiliensis
Authors: Almeida Igor C. , Rodrigues Elaine G. , Travassos Luiz R. 
Published: March 2007

AbstractQuantitative chemiluminescent enzymelinked immunosorbent assay (ELISA) and dot‐blotting procedures were developed to evaluate the reactivity of human antibodies with crude antigens and purified molecules of parasites and fungi, mainly Trypanosoma cruzi and Paracoccidioides brasiliensis. Reproducible, highly sensitive, and strictly doseresponding results were obtained, with the specificity depending on the kind of antigen used. Mixed antigens (epimastigote membrane and HIV‐1 heptapeptide) applied in dots could be independently recognized by specific sera. Purified antigens (T. cruzi F2/3 and P. brasiliensis gp43) at very small concentrations gave specific reactions with patients' sera diluted ≥1:1,000 and were very poorly reactive or unreactive with natural antibodies using the chemiluminescent immunoassays. P. brasiliensis crude antigen Fava Netto polysaccharide antigen (FNPA) contained peptide epitopes recognized by natural antibodies and carbohydrate epitopes reactive with sera from histoplasmosis patients. It is very important that sensitive chemiluminescence immunoassays be used with purified antigenic molecules to ensure specificity for the diagnosis and follow‐up of parasitic and fungal infections. © 1994 Wiley‐Liss, Inc.
Enzymatic fluorometric assay for tissue cAMP
Authors: Sugiyama Atsushi , Wiegn Phi , McKnite Scott , Lurie Keith G. 
Published: March 2007

AbstractcAMP is commonly measured using either immunoassay or high‐performance liquid chromatography. The current methods are sensitive but may lack versatility and be expensive; also, radioactivity is potentially harmful to the operator and environment. Given these concerns, we developed a highly sensitive enzymatic fluorometric assay for cAMP. The method consists of five steps: (1) destruction of interfering compounds with apyrase, 5′ nucleotidase, adenosine deaminase, and alkaline phosphatase; (2) conversion of cAMP to AMP; (3) conversion ofAMP to ATP; (4) amplification ofATP by ATP‐ADP cycling; and (5) fluorometric measurement of resultant NADPH. cAMP was measured in male Sprague Dawley rats anesthetized with pentobarbital. Stimulated rats (n = 4) received isoproterenol (16 μg/kg, s.q.) and aminophylline (20 mg/kg, s.q.), whereas controls (n = 4) received no additional drug. With the enzymatic fluorometric assay, cAMP content in heart, liver, and kidney (pmol/mg wet wt, mean ± SEM) was 0.34 ± 0.03, 0.33 ± 0.03, and 0.92 ± 0.11 in the control group and 0.77 ± 0.10, 0.66 ± 0.04, and 1.53 ± 0.12 in the stimulated group, respectively. The total assay duration including sample reading procedure varied at 4.5–9.5 hr, depending on its sensitivity. cAMP from the same samples was measured using a commercially available enzyme immunoassay kit and was found to be very similar to the enzymatic fluorometric assay. We conclude that this new assay is sensitive, safe, versatile, and inexpensive and can be used to measure cAMP in multiple types of tissue, including biopsy samples weighing <200 μg. © 1994 Wiley‐Liss, Inc.
Rapid method for purification of CD56+ natural killer cells with preferential enrichment of the CD56bright+ subset
Authors: Handgretinger Rupert , Welte Birgitta , Dopfer Roland , Niethammer Dietrich 
Published: March 2007

AbstractA rapid method for the purification of CD56+ natural killer (NK) cells with the preferential enrichment of the CD56bright+ subset is described. The method is based on the adsorption of CD56+ cells indirectly stained with a biotinylated antibody on an avidin‐coated column. The adsorbed CD56+ cells can then be squeezed out mechanically without damaging the viability or the function of the cells. Starting from peripheral mononuclear cells from adult donors, the recovery of the CD56+ cells was 11.9 ± 9.0% (n = 8), the purity 93.0 ± 4.9% (n = 10), and the enrichment factor 7.5 ± 1.7 (n = 8). Further phenotypic classification of the CD56+ cells into CD56dim+ and CD56bright+ cells showed a preferential enrichment of the CD56bright+ phenotype with a recovery of 40.5 ± 19.6% (n = 8), a purity of 30.3 ± 13.2% (n = 8), and an enrichment factor of 29.8 ± 7.2 (n = 8). In conclusion, the described method allows the rapid purification of CD56+ NK cells with the preferential enrichment of the CD56bright+ subset. © 1994 Wiley‐Liss, Inc.
Laboratory study on occult gastrointestinal bleedings: Differential diagnostic method for the parallel study of intact and fragmented hemoglobin
Authors: Ottó Szabolcs , Németh Mária , Kocsis György 
Published: March 2007

AbstractThe authors produced various degradation products of human hemoglobin. With the aid of immune sera raised against native hemoglobin and their fragments, we examined the antigenicity of the intact molecule and fragments as well as the specificity and reactivity of the immune sera used. Since conventional guaiac‐type reactions make no distinction between the different hemoglobin molecules and the methods described in recent years' publications only with immune sera raised against native intact hemoglobin, the demonstration of occult fecal blood has not reached the desired rate of efficacy in clinical practice. The authors recommend the introduction of a simple differential diagnostic method suitable for the parallel examination of intact and fragmented hemoglobin.©1995 wiley‐Liss, inc.
Decreased cytoplasmic immunoglobulin a production during epstein‐barr virus immortalization on lymphocytes from patients with ataxia‐telangiectasia
Authors: Okano Motohiko , Sakiyama Yukio , Matsumoto Shuzo , Mizuno Fumio , Osato Toyoro 
Published: March 2007

AbstractPreviously, we have reported significantly lower immunoglobulin (Ig) A production in supernatants of cultured lymphoblastoid cells using enzyme‐linked immunosorbent assay from patients with ataxia‐telangiectasia (AT) when compared to that of age‐ and sex‐matched healthy individuals. Here, we further assess the degree of cytoplasmic Ig production in these cells and also analyze it during the early phase of Epstein‐Barr virus immortalization. All classes of cytoplasmic IgM, IgG, and IgA productions were demonstrated in cells from healthy controls. In contrast, cells from patients with AT showed only cytoplasmic IgM and IgG with low or nondetectable levels of IgA during and after the immortalizing process. These results suggest B lymphocytes bearing IgA are functionally immature and/or defective in patients with AT.©1995 wiley‐Liss, inc.
Quantification of mitogen induced human lymphocyte proliferation: Comparison of alamarbluetm assay to 3h‐thymidine incorporation assay
Authors: De Fries Ricarda , Mitsuhashi Masato 
Published: March 2007

AbstractProliferation of human lymphocytes in response to various stimuli has traditionally been assessed by measuring uptake of radiolabeled nucleotides such as 3H‐thymidine. We have evaluated a fluorometric assay, which uses the commercially available reagent, alamarBlue, as a potential substitute for the 3H‐thymidine assay in measuring proliferation of human lymphocytes. In this assay, alamarBlueTM is added to a population of cells where it is reduced by mitochondrial enzyme activity. The reduced form of the reagent is fluorescent and can be quantitatively detected. The safety and convenience of the alamarBlueTM assay make it very attractive for use in the clinical laboratory.In this study peripheral blood mononuclear cells (PBMC) from healthy donors were stimulated using the mitogen Concanavalin A, and proliferation was assessed using either the 3H‐thymidine or the alamarBlueTM assay. The alamarBlueTM assay reliably detects human PBMC and we found that the linear range of detection was 104 cells/well (96‐well plate) to 5 × 105 cells/well. Detection of human PBMC is highly reproducible and the alamarBlue assay may be suitable in a number of applications where detection or relative quantitation of human PBMC is required. The alamarBlue assay also detected mitogen induced proliferation of PBMC although with a significantly lower level of sensitivity than the standard 3H‐thymidine assay.©1995 wiley‐Liss, inc.
Detection of (1–3)‐β‐D‐glucan in a rat model of aspergillosis
Authors: Mitsutake Kotaro , Kohno Shigeru , Miyazaki Takashige , Yamamoto Yoshihiro , Yanagihara Katsunori , Kakeya Hiroshi , Hashimoto Atsuro , Koga Hironobu , Hara Kohei 
Published: March 2007

AbstractThe G test containing factor G, fractioned from the Limulus lysate, was used to detect (1–3)‐β‐D‐glucan in a rat model of aspergillosis. Aspergillus fumigatus strain MF‐13, 1 × 104 conidia, were inoculated transtracheally into rats treated with cortisone acetate (100 mg/kg) and fed a low‐protein (8%) diet. Increased serum (1–3)‐β‐D‐glucan was found on the sixth day after inoculation in concentrations of 370 ± 178 pg/ml (mean ±SD) in untreated controls, and 154 ± 43 pg/ml in rats treated with 0.5 mg/kg of amphotericin B. On day 11 (1–3)‐β‐D‐glucan concentrations were 2,590 ± 2,940 pg/ml and 448 ± 442 pg/ml, respectively. The elevation in levels of (1–3)‐β‐D‐glucan increased in correlation with the elevation of galactomannan antigen titers; (1–3)‐β‐D‐glucan is thus measurable during experimental aspergillosis in rats.©1995 wiley‐Liss, inc.
Prostate‐Specific antigen expression by various tumors
Authors: Levesque Michael , Hu He , Diamandis Eleftherios P. , D'Costa Mario 
Published: March 2007

AbstractThere is a growing body of evidence indicating that prostate‐specific antigen (PSA) may be present in many steroid hormone‐stimulated epithelial tissues other than that of the prostate. In particular, breast tumor cell lines treated with steroid hormone receptor agonists, breast tumors, and normal human breast have recently been found by our group to contain PSA. To investigate whether PSA may also be present in other human tumors, we employed a highly sensitive immunofluorometric assay technique to quantify PSA immunoreactivity in tumor extracts. Using a PSA‐positivity cutoff value of 0.005 ng per mg of protein, 23 of 43 diverse tumors tested positive for PSA protein. Confirmatory analyses for PSA by a commercially available method (IMx) on six samples demonstrated a high degree of concordance between the two methods. To establish the molecular weight of the immunoreactive species, the most highly positive tumor extracts of each tumor type were fractionated by high performance liquid chromatography. Whereas the majority of tumors had immuoreactivity eluting at both 100 KDa and 33 KDa, corresponding to PSA bound to α1‐antichymotrypsin and free PSA, respectively, the colon and parotid tumors displayed immunoreactivity only at the 33 KDa fraction. We conclude that in addition to breast tumors and normal breast, colon, ovarian, liver, kidney, adrenal, and parotid tumors can also produce PSA. The physiological role of PSA in these tumors is currently under investigation.©1995 wiley‐Liss, inc.
Evaluation of a new automated system for the determination of ck‐mb isoforms
Authors: Secchiero Sandra , Altinier Sara , Zaninotto Martina , Lachin Mattia , Plebani Mario 
Published: March 2007

AbstractWe evaluated a new analyzer (Cardio REP) specifically designed for cardiac CK‐MB isoenzyme and isoforms activity, with a performance time of 24 minutes. Ten AMI patients, with times elapsed between the onset of chest pain and admission to hospital ranging from 30 minutes to 4 hours, were monitored every 3–4 hours until the 16th hour of hospitalization. In each serum sample, in addition to total CK‐MB and CK‐MB isoforms measured by the Cardio REP analyzer, we also assayed total CK activity, CK‐MB activity by immunoinhibition method, CK‐MB mass concentration, CK‐MB isoforms by REP method, troponin T, and myoglobin. The precision study demonstrated acceptable within assay and between assay CVs% for total CK‐MB (8.1 and 10.4), MB1 (9.1 and 14.2), and MB2 (9.1 and 8.2) isoforms. The method was found to be linear up to 371 U/L for MB2 isoform fraction and up to 516 U/L for total CK‐MB. Results for CK‐MB obtained with the Cardio REP correlated well with those for CK‐MB activity obtained with the immunoinhibition method (r = 0.869) and those of CK‐MB mass concentration (r = 0.923). The sensitivity of the Cardio REP CK isoforms method was found to be greater than that of the REP CK isoforms method. Time to first increased value of MB2/MB1 ratio and MB2 isoform was earlier in comparison to that for CK‐MB mass concentrations and similar to that for myoglobin, a marker that, however, lacks specificity. The diagnostic efficiency of CK‐MB isoforms and the availability of a real‐time, fully automated method for their measurement suggest the utilization of this biochemical marker in emergency for the early diagnosis of AMI.
Utilization of monoclonal‐antibody‐based assay (hemocardtm) in screening for and differentiating between genotypes of sickle cell disease and other hemoglobinopathies
Authors: Schultz John C. 
Published: March 2007

AbstractSickle cell disease covers a group of conditions in which pathology may be attributed to the presence of sickle hemoglobin (HbS). The identification of HbS and other variants including those in combination with HbS is commonly achieved by cellulose acetate electrophoresis at alkaline pH. Because many hemoglobin variants with similar charges have similar electrophoretic migration patterns, they are difficult to differentiate by electrophoresis. The HemoCardTM assays address this concern through the use of monoclonal antibodies capable of specifically recognizing the unique amino acid sub‐stitution in the variant hemoglobin. The panel of HemoCard monoclonal antibodies confirms the absence and presence of HbA, HbC, HbE, HbS, and other sickling hemoglobin variants. The combination of alkaline cellulose acetate electrophoresis and HemoCard assays allows the technologist to reach a final confirmation of both common and much less common sickle cell disease genotypes, combinations of HbS with other hemoglobins that ordinarily do not produce sickle cell disease, and other clinically important hemoglobinopathies including HbE/ß‐thalassemia and hemoglobin C disease.
Multilayer peroxidase‐labeled antibody method: Comparison with labeled streptavidin‐biotin method, avidin‐biotin‐peroxidase complex method, and peroxidase‐antiperoxidase method
Authors: Shin Masashi , Izumi Shin‐Ichi , Nakane Paul K. 
Published: March 2007

AbstractA new, extremely sensitive, “multilayer peroxidase‐labeled antibody method” is introduced. This method utilizes peroxidase‐labeled antibodies that immuno‐react with their antigen, but also can be an antigen to other peroxidase‐labeled antibodies. An example of this method is: use rabbit IgG as the first antibody, peroxidase‐labeled goat antirabbit IgG as the second antibody, peroxidase‐labeled rabbit antigoat IgG as the third antibody, peroxidase‐labeled goat anti‐rabbit IgG as the fourth antibody, and peroxidase‐labeled rabbit antigoat IgG as the fifth antibody in succession. We compared the sensitivities of this multilayer peroxidase‐labeled antibody method with that of commercially available immuno‐peroxidase staining kits, i.e., avidin‐biotin‐complex method and peroxidase‐antiperoxidase method, both by immunostaining and enzyme‐linked immunosorbent assay. The generated signal and background in immunostained sections were determined by a computer‐assisted image analyzer and by enzyme‐linked immunosorbent assay spectrophotometrically. It was found that a maximum signal/background ratio was obtained with those sections reacted with the first four antibodies in succession by the multilayer peroxidase‐labeled antibody method, and those reacted with the five antibodies in succession by the multilayer peroxidase‐labeled antibody method in the enzyme‐linked immunosorbent assay. Thus, the multilayer peroxidase‐labeled antibody method is more sensitive than the avidin‐biotin‐complex method and the peroxidase‐antiperoxidase method.