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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 92 (1989), S. 397-406 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have produced a small library of colonic mucosa and colorectal carcinoma reactive monoclonal antibodies (MoAbs) by immunizations with extracts of human colon cancer tissue and a human colon cancer cell line. Hybridoma supernatants were tested on (normal and neoplastic) human tissues by immunoperoxidase methods to evaluate organ or tissue specificity. Initial biochemical characterization of the target antigens was performed by gelpermeation chromatography, Western blotting and competition assays. Based upon the immunoreactivity patterns and the characteristics of the antigen four groups of MoAbs could be distinguished. The first group concerns the antibodies PAR-LAM 3, 9 and 10. These antibodies react with an 87 kDa protein moiety in high molecular weight (2–5×106 Da) glycoproteins. In intestinal and colon mucosa these antibodies showed diffuse binding with goblet cells. In colon carcinoma decreased reactivity with these MoAbs was found. The second group consists of antibodies PARLAM 8, 12 and 13. These antibodies react with large (〉5×106 Da) glycoproteins, most likely with carbohydrate epitopes. By immunohistochemistry in normal colon mucosa the antibodies all show granular supranuclear reactivity with goblet cells. These antibodies show increased reactivity with colon adenomas and adenocarcinomas. A third group is formed by PARLAM 2, which also reacts with a large (〉5×106 Da) glycoprotein, showing a granular distribution in goblet cells. In colon carcinomas more extensive expression is found than in normal colonic mucosa. Finally, the fourth group consists of PARLAM 11, which also reacts with a large (〉5×106 Da) glycoprotein, located in the brush border of colonic columnar cells. These antibodies might be useful tools for the analysis of the expression of mucin related glycoproteins in normal, preneoplastic and neoplastic colon mucosa.
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  • 2
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Ataxia telangiectasia (AT) is an autosomal recessive disorder characterized by cerebellar ataxia, telangiectasia, immunodeficiency, elevated α-fetoprotein levels, chromosomal instability, predisposition to cancer, and radiation sensitivity. We report the identification of a new, double missense mutation in the ataxia telangiectasia gene (ATM) of a Dutch family. This homozygous mutation consists of two consecutive base substitutions in exon 55: a T→G transversion at position 7875 of the ATM cDNA and a G→C transversion at position 7876. These transversions were confirmed by polymerase chain reaction/primer-induced restriction analysis with CelII. The double base substitution results in an amino acid change of an aspartic acid to a glutamic acid at codon 2625 and of an alanine to a proline at codon 2626 of the ATM protein. Both amino acids are conserved between the ATM protein and its functional homolog, the Atm gene product in the mouse. Furthermore, the Chou-Fasman and Robson predictions both demonstrate a change in the secondary structure of the ATM protein carrying the D2625E/A2626P mutation. These findings suggest that the double base substitution in the ATM gene is a disease-causing mutation.
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  • 3
    ISSN: 1434-6052
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract Inelastic and elasticJ/ψ (3097) photoproduction on Li6 are measured at a mean γ energy of 90 GeV in an open spectrometer. TheJ/ψ are identified by their decays intoμ + μ − ore + e −. A signal of ψ′(3685) intoμ + μ − andJ/ψπ + π − is also seen. The inelastic cross-section withZ=E ψ/Eγ〈0.9 is compared in shape and magnitude with the colour singlet model of photon-gluon fusion.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Journal of applied electrochemistry 25 (1995), S. 122-125 
    ISSN: 1572-8838
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Electrical Engineering, Measurement and Control Technology
    Notes: Abstract Electrodes for air-driven PEMFCs for transport applications have been developed. The structure of the electrodes has been specifically adapted to run with air as oxidant under near atmospheric pressure; such electrodes can be manufactured using conventional industrial methods and be easily scaled up. The technology has been demonstrated on a 50 cm2 electrode area, assembled together with a Nafion® 117 membrane. Electrodes with different platinum loading, namely 0.4 and 0.2 mg Pt cm−2, have been the subject of long duration tests which show a slow degradation of the cell performance. With air as oxidant at 180 kPa absolute pressure, 80°C as cell temperature and Nafion® 117 as membrane, a power density of 350 mW cm−2 has been obtained.
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  • 5
    ISSN: 1573-5028
    Keywords: Genetic engineering ; KDEL retention signal ; signal peptide ; single-chain antibody ; targeting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of subcellular localization on single-chain antibody (scFv) expression levels in transgenic tobacco was evaluated using an scFv construct of a model antibody possessing different targeting signals. For translocation into the secretory pathway a secretory signal sequence preceded the scFv gene (scFv-S). For cytosolic expression the scFv antibody gene lacked such a signal sequence (scFv-C). Also, both constructs were provided with the endoplasmic reticulum (ER) retention signal KDEL (scFv-SK and scFv-CK, respectively). The expression of the different scFv constructs in transgenic tobacco plants was controlled by a CaMV 35S promoter with double enhancer. The scFv-S and scFv-SK antibody genes reached expression levels of 0.01% and 1% of the total soluble protein, respectively. Surprisingly, scFv-CK transformants showed considerable expression of up to 0.2% whereas scFv-C transformants did not show any accumulation of the scFv antibody. The differences in protein expression levels could not be explained by the steady-state levels of the mRNAs. Transient expression assays with leaf protoplasts confirmed these expression levels observed in transgenic plants, although the expression level of the scFv-S construct was higher. Furthermore, these assays showed that both the secretory signal and the ER retention signal were recognized in the plant cells. The scFv-CK protein was located intracellularly, presumably in the cytosol. The increase in scFv protein stability in the presence of the KDEL retention signal is discussed.
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  • 6
    ISSN: 1573-5028
    Keywords: carrot ; cell wall ; suspension culture ; secreted glycoprotein ; root ; nodulin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A 47 kDa glycoprotein, termed EP4, was purified from carrot cell suspension culture medium. An antiserum raised against EP4 also recognized a protein of 45 kDa that was ionically bound to the cell wall. EP4 was detected in culture media from both embryogenic and non-embryogenic cell lines and was found to be secreted by a specific subset of non-embryogenic cells. Secretion of the 47 kDa glycoprotein by embryogenic cells was not evident. The 45 kDa cell wall-bound EP4 protein was specific for non-embryogenic cells and was shown by immunolocalization to occur in the walls of clustered cells, with the highest levels in the walls separating adjacent cells. In seedlings, EP4 proteins were mainly found in roots. EP4 cDNA was cloned by screening a cDNA library with an oligonucleotide derived from an EP4 peptide sequence. The EP4 cDNA sequence was found to be 55% homologous to ENOD8, an early nodulin gene from alfalfa.
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  • 7
    ISSN: 1573-5028
    Keywords: complementation ; expression ; co-suppression ; granule-bound starch synthase ; Solanum tubersoum L. ; transgenic inheritance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The wild-type gene encoding granule-bound starch synthase (GBSS) is capable of both complementing the amylosefree (amf) potato mutant and inhibiting the endogenous GBSS gene expression in wild-type potato. Co-suppression of the endogenous GBSS gene, easily visualised by staining the starch with iodine, occurred when the full-size GBSS sequence (genomic), GBSS cDNA or even the mutant amf allele were introduced into the wild-type potato. Conversely, introduction of the GBSS promoter sequence alone, did not result in co-suppression in the 80 analysed transformants. Neither the orientation of the GBSS gene with respect to kanamycin resistance nor the presence of an enhancer influenced the frequency of plants showing a co-suppression phenotype. After crossing a partially complemented amf mutant with a homozygous wild-type plant, the F1 offspring segregated into plant phenotypes with normal and decreased expression of the GBSS gene. This decreased expression correlated with the presence of a linked block of five T-DNA inserts which was previously shown to be correlated with partial complementation of the amf mutant. This crossing experiment indicates that co-suppression can cause inhibition of gene expression of both inserted and endogenous wild-type GBSS genes. The frequency of partially complemented amf plants was equal to the frequency of co-suppressed wild types when a construct, with an enhancer in front of the GBSS promoter, was used (pWAM 101E). This might suggest that partial complementation of the amf genotype caused by unstable expression of the transgene can be overcome by inserting an enhancer in front of the GBSS promoter.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Environmental monitoring and assessment 34 (1995), S. 203-214 
    ISSN: 1573-2959
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Cellular automata provide the key to a dynamic modelling and simulation framework that integrates socio-economic with environmental models, and that operates at both micro and macro geographical scales. An application to the problem of forecasting the effect of climate change on a small island state suggests that such modelling techniques could help planners and policy makers design more effective policies — policies better tuned both to specific local needs and to overall socio-economic and environmental constraints.
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  • 9
    ISSN: 1862-0760
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract In the eighties, Iwahara discovered that doped Sr and Ba cerates show significant proton conduction, at high temperature, and in hydrogen- or water-containing atmosphere [1]. Begin ‘90 it was proved that also doped zirconates (Sr, Ba, Ca) join this family of proton conducting perovskites [2]. Within this group, indium doped CaZrO3 is one of the more interesting materials for applications, being mechanically and chemically particularly stable, and producible in very dense form. Specifically, it was already succesfully incorporated in a hydrogen sensor for liquid aluminium [3]. In principle, this sensor works as a simple electrochemical hydrogen concentration cell described by Nernst. However, as already remarked phenomenologically from the beginning [1], a hydrogen concentration cell using these perovskites shows the theoretically predicted emf, only in the presence of humidity. In this paper we want to go deeper into the reasons for this phenomenon.
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  • 10
    ISSN: 1573-9368
    Keywords: transformation vector ; pBIN19 ; transgenic plants ; T-DNA ; Agrobacterium tumefaciens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We describe the construction of a new plant transformation vector, pBINPLUS, based on the popular pBIN19 vector. Improvements over pBIN19 include location of the selectable marker gene at the left T-DNA border, a higher copy number inE. coli, and two rare restriction sites around the multiple cloning site for easier cloning and analysis of T-DNA insertions in plant genomes.
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