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  • 1
    Keywords: TUMORS ; MICE ; ACTIVATION ; MOUSE ; ACUTE LYMPHOBLASTIC-LEUKEMIA ; CANCER-CELLS ; HALLMARKS ; APOPTOSOME
    Abstract: AVEN has been identified as an inhibitor of apoptosis, which binds to the adaptor protein, APAF-1, and thereby prevents apoptosome formation and mitochondrial apoptosis. Recent data have demonstrated high expression levels of AVEN messenger RNA in acute leukemias as well as a positive correlation between AVEN mRNA overexpression and poor prognosis in childhood acute lymphoblastic leukemia. On the basis of these data, we investigated the potential involvement of AVEN in tumorigenesis. First, we confirmed the overexpression of AVEN in T-cell acute lymphoblastic leukemia/lymphoma (T-ALL) patient samples. We then established a transgenic mouse model with T-cell-specific overexpression of AVEN, with which we demonstrated the oncogenic cooperation of AVEN with heterozygous loss of p53. Finally, we used a subcutaneous xenograft mouse model to show that AVEN knockdown in the T-ALL cell lines, MOLT-4 and CCRF-CEM, and in the acute myeloblastic leukemia cell line, Kasumi-1, leads to a halt in tumor growth owing to the increased apoptosis and decreased proliferation of tumor cells. Collectively, our data demonstrate that the anti-apoptotic molecule, AVEN, functions as an oncoprotein in hematopoietic neoplasms.Oncogene advance online publication, 2 July 2012; doi:10.1038/onc.2012.263.
    Type of Publication: Journal article published
    PubMed ID: 22751129
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  • 2
    Keywords: PEPTIDE ; QUANTIFICATION ; microarrays ; DESIGN ; IMMUNOASSAYS ; Antibody detection ; MYCOBACTERIAL LIPOARABINOMANNAN ; single-molecule sensitivity
    Abstract: Based on a single-molecule sensitive fluorescence-linked immunosorbent assay, an analytical platform for the detection of lipoarabinomannan (LAM), a lipopolysaccharide marker of tuberculosis, was established that is about 3 orders of magnitude more sensitive than comparable current ELISA assays. No amplification step was required. Also, no particular sample preparation had to be done. Since individual binding events are detected, true quantification was possible simply by counting individual signals. Utilizing a total internal reflection configuration, unprocessed biological samples (human urine and plasma) to which LAM was added could be analyzed without the requirement of sample purification or washing steps during analysis. Samples containing about 600 antigen molecules per microliter produced a distinct signal. The methodology developed can be employed for any set of target molecules for which appropriate antibodies exist
    Type of Publication: Journal article published
    PubMed ID: 21247063
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  • 3
  • 4
    Keywords: APOPTOSIS ; CELLS ; IN-VITRO ; BLOOD ; CELL ; Germany ; INHIBITION ; DISEASE ; HEART ; PATIENT ; MACROPHAGES ; SERA ; treatment ; ACID ; GLUTATHIONE ; PLASMA ; DECREASE ; cholesterol ; LDL ; LIPOPROTEIN ; PERIPHERAL-BLOOD ; OXIDATION ; cysteine ; arteriosclerosis,risk factors in hyperlipidemia,glutathione in atherosclerosis,redox status as a ris ; CORONARY-ARTERY DISEASE ; HEART-DISEASE ; N-ACETYL-CYSTEINE ; SERUM LEVELS
    Abstract: Treatment of hyperlipidemic patients with the thiol compound N-acetyleysteine (NAC) was previously shown to cause a significant dose-related increase in the high-density lipoprotein (HDL) -cholesterol serum level, suggesting the possibility that its disease-related decrease may result from a diminished thiol concentration and/or thiol/disulfide redox status (REDST) in the plasma. We therefore investigated plasma thiol levels and REDST in normo-/byperlipidemic subjects with and without coronary heart disease (CHD). The thiol level, REDST, and amino acid concentrations in the plasma and intracellular REDST of peripheral blood mononuclear cells (PBMC) have been determined in 62 normo- and hyperlipidemic subjects. Thirty-three of these subjects underwent coronary angiography, because of clinical symptoms of CHD. All groups of hyperlipidemic patients under test and those normolipidemic individuals with documented coronary stenoses showed a marked decrease in plasma thiol concentrations, plasma and intracellular REDST of PBMCs, and a marked increase in plasma taurine levels. Individual plasma thiol concentrations and plasma REDST were strongly negatively correlated with the serum LDL-cholesterol and positively correlated with the serum HDL-cholesterol level. Together with the earlier report about the effect of NAC on the HDL-cholesterol serum level, our findings suggest strongly that lower HDL-cholesterol serum levels may result from a decrease in plasma thiol level and/or REDST possibly through an excessive cysteine, catabolism into taurine. (C) 2003 Elsevier Inc
    Type of Publication: Journal article published
    PubMed ID: 14607527
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  • 5
    ISSN: 1573-904X
    Keywords: Fab fragment ; radioimmunoassay ; pharmacokinetics ; urinary metabolites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Anti-sheep Fab fragment antisera were produced in rabbits using sheep digoxin-specific Fab fragments (Digidot) as immunogen. These antisera were used for the development of a radioimmunoassay (RIA) of sheep Fab fragments in human plasma and urine using 125I-labeled Fab fragments. Interference in the assays by digoxin, human proteins, and antibodies from different species was insignificant, but cross-reactivity between anti-sheep Fab antisera and goat IgG or Fab fragments was 22 to 67%. The limit of detection was 0.1 µg/mL and the assay was linear over a 0.6–28 µg/mL range of Fab fragments. Intra- and interassay coefficients of variation were less than 6.9 and 10.5%, respectively. Accuracy of plasma and urine assays at various Fab fragment levels ranged from 96 to 106%. RIA was applied to the pharmacokinetic study of sheep digoxin-specific Fab fragments in one patient acutely intoxicated by digitoxin and treated with Digidot. The Fab elimination half-life was 12.1 hr. Steady-state volume of distribution and total-body clearance were 10.8 L and 23.4 mL/min, respectively. Unchanged Fab fragments (50 kD) and degradation products (25 kD) isolated by gel filtration chromatography of a urine sample cross-reacted with the anti-Fab antiserum.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-6822
    Keywords: genotoxicity ; UDS ; hepatocytes ; attapulgite ; xonotlite ; sepiolite
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The biological activity of natural and synthetic mineral fibers has been examined. Natural attapulgite [(Mg, Al)2Si4O10(OH).4H20], synthetic xonotlite [Ca3Si3O8(OH)2] and natural sepiolite [Mg2Si3O8.2H2O] were selected. Genotoxic effects were investigated by means of a well established cellular model based upon the measurement of unscheduled DNA synthesis (UDS) in rat hepatocytes in primary culture. The intrinsic capacity of the fibers (1 and 10 µ/ml) to induce UDS was first tested. None of the fiber types showed detectable UDS-eliciting activity. Also, the possible modulation of the cellular response to genotoxic agents by the materials was examined by exposing the cells to mixtures of 2-acetylaminofluorene (AAF) (0.05 and 0.25 µg/ml) and fibers (1 and 10 µg/ml). In these experiments, the UDS response was significantly diminished in the presence of xonotlite. This phenomenon may reflect changes in the uptake and/or metabolism of AAF or may result from an inhibition of DNA repair processes, the latter suggesting a possible cocarcinogenic potential for this synthetic silicate. These results point to the immediate necessity of studying more extensively the biological effects of fibrous materials that can be used as substitutes for asbestos.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-904X
    Keywords: digoxin ; monoclonal antibodies ; redistribution ; immunoreactivity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The effect of three monoclonal digoxin-specific antibodies on total and free digoxin plasma disposition was studied in rats in order to determine the role of affinity constant (Ka) and dose. Thirty minutes after digoxin infusion, administration of a stoichiometrical dose of the 1CIO, 6C9 and 9F5 IgG (Ka = 6 109, 3.1 108 and 2.5 107 M−1, respectively) resulted in a plasma digoxin increase linearly related to Ka. The mean free plasma digoxin was 0.6 ± 0.4, 7.8 ± 3.3 and 43 ± 22 % respectively after 1C10, 6C9, and 9F5 IgG infusion in comparison to 70 ± 9% in the control group. When the IgG:digoxin ratio increased from 1 to 5, plasma digoxin Cmax and AUCT also increased as a function of both affinity (Ka) and dose (N), but not linearly. The product of NKa defined an immunoreactivity factor that was well fitted to the digoxin redistribution parameters (Cmax and AUCT) by a Hill equation.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-904X
    Keywords: immunoreactivity ; IgG and Fab fragments ; colchicine ; digoxin ; digitoxin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract This study investigates immunoreactivity control procedures, i.e., specificity, affinity constant (K a ), and specific active binding sites (SABS), for polyclonal anticolchicine, monoclonal antidigitoxin IgG and Fab fragments, and antidigoxin Fab fragments (Digidot). Preliminary control procedures for IgG and Fab fragment purity indicated that all reagents were immunologically pure. All IgG and Fab fragments exhibited similar cross-reactivity and K a. No decrease in percentage of Fab fragment SABS was observed after papain cleavage of anticolchicine and antidigitoxin IgG. Nevertheless, only 4.3 ± 1.2% of nonimmunopurified anticolchicine polyclonal Fab fragments and 76.2 ± 2.3 to 88.7 ± 2.5% of different batches of immunopurified anti-digoxin Fab (Digidot) were active, the latter percentage being in the range of the 85% specified by the manufacturer. Only 58 ± 3% of digitoxin-specific monoclonal IgG was active and 67 ± 7% of its Fab fragments. Results show the importance of determining the ratio of SABS to presumed total specific binding sites for pharmaceutical monoclonal and polyclonal antibody preparations against haptens.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1573-904X
    Keywords: monoclonal antibody ; hapten ; specificity ; affinity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract This review deals with the potency of monoclonal antibodies (MAbs) to haptens in immunoassays. Specificity and affinity of MAbs to haptens are the major determinants to be considered. Specificity of MAbs depends on the selection of the hapten coupling site to the carrier protein and the antigen used for the screening of MAbs. Nevertheless, cross-reactivity can occur with compounds related to the hapten. This poly specificity may be circumvented with the use of many MAbs, as has been demonstrated for MAbs to cyclosporine. Affinity of MAbs to haptens is often lower than that of corresponding polyclonal antibodies (PAbs), thereby limiting assay sensitivity. Low affinity is more frequently observed with low molecular weight (100–300) haptens than with larger haptens, such as digoxin or cyclosporine. Affinity enhancement by increasing resemblance to the immunogen can be effective in resolving the lack of sensitivity. With suitable selection strategies, MAbs exhibit real advantages over classical PAbs to haptens because large amounts of worldwide standardized reagents can be prepared.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1618-2650
    Keywords: Elementaranalyse, Datenverarbeitung ; Organisation eines Labors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Description / Table of Contents: Zusammenfassung Es wird das Konzept eines Echtzeit-Datenverarbeitungssystems beschrieben, das in Kombination mit mechanisierten und automatisierten Analysenmethoden eine Verbesserung der analytischen Serviceleistung ermöglicht. Die Hardware setzt sich im wesentlichen aus einem 28 K-Worte Kleincomputer mit 2 Plattenspeichereinheiten, Bedienungskonsolen und Beleglesern zusammen. Die hauptsächlichen Funktionen dieses Teils der Anlage sind: Registrieren der Aufträge, Erstellen von Arbeitslisten für die Laboratorien, Überwachen der termingerechten Auftragserledigung, Sammeln, Speichern und Auswerten der anfallenden Resultate, Treffen einfacher Entscheidungen und schließlich Erstellen und Ausdrucken der Analysenbefunde. Darüber hinaus verfertigt der Computer täglich Berichte über den Stand der Auftragserledigung und speichert Daten für die Statistiken und Kostenabrechnungen. Zur Datenübertragung aus den Laboratorien stehen 36 Linien zur Verfügung. Dabei ist wesentlich, daß elektronisch vorverarbeitete Daten, also fertig berechnete Resultate übertragen werden. Dazu dienen on-line angeschlossene Tischrechner, Datenstationen und Belegleser sowie Datenverarbeitungseinheiten (Prozent-Box) eigener Konstruktion. Die Arbeitsplätze in den Laboratorien sind somit weitgehend autonom. Dieses hierarchische Datenverarbeitungssystem bringt arbeitstechnische Vorteile, gewährleistet aber vor allem die ständige Servicebereitschaft unserer Laboratorien, da auch bei Ausfall des Computers an den Analysengeräten unbeeinflußt weitergearbeitet werden kann.
    Notes: Abstract To increase the efficiency of the analytical services the analytical methods have been improved by mechanisation and automation, and combined with a real-time data processing system. The hardware consists essentially of a 28 K-words minicomputer with 2 disc storage units, teletypes and optical mark readers. The main functions of this part of the system are as follows: registering the incoming requests, supplying work schedules for the various laboratories, controlling the completion of the work within the required time, collecting, storing, and processing the results, making simple decisions, and finally, editing and printing the analysis reports. Moreover, the computer produces daily reports on the state of progress of requests, and stores the data for usage in statistics and cost accounting. There are 36 lines available for the transfer of data from the laboratories. An important feature of the system design is the transfer of electronically preprocessed data, i.e. already calculated final results. For this purpose, on-line desk-top calculators, alphanumeric display data terminals, and data preprocessing units (percentage-box) designed and developed in our own laboratories are being used. The individual places of work in the laboratories are thus to a great extent autonomous. This hierarchical data processing system yields technical advantages, and most of all, guarantees a permanent service ability of our laboratories since even in the case of a breakdown of the computer, the analytical work can be continued without any interference.
    Type of Medium: Electronic Resource
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