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  • 1
    Electronic Resource
    Electronic Resource
    Woodbury, NY : American Institute of Physics (AIP)
    Applied Physics Letters 73 (1998), S. 3698-3699 
    ISSN: 1077-3118
    Source: AIP Digital Archive
    Topics: Physics
    Notes: To identify Ga- or Zn-related deep levels, deep level transient spectroscopy (DLTS) measurements were performed repeatedly during the elemental transmutation of 67Ga to 67Zn. The radioactive isotope 67Ga was recoil implanted into p-type 6H–SiC for radiotracer experiments. The DLTS spectra exhibit one peak of time-dependent height. It describes the increasing concentration of the daughter element Zn with the half life of the nuclear decay. Thus, one Zn-related level at 1.16 eV above the valence band edge is definitely identified. There is no deep level of Ga in the lower part of the band gap. © 1998 American Institute of Physics.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1600-0560
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Monoclonal antibody T311 specifically detects tyrosinase protein expression. Tyrosinase-derived peptides are recognized by CD8+ T-celis and applied in immunotherapy. We examined formalin-fixed paraffin-embedded tissue of 50 melanoma (primary n=31, metastatic n=19) and 41 control cases (junctional, dermal, compound, Spitz, Reed, balloon-cell nevi) by immunochemistry using the alkaline phosphatase-anti-alkaline phosphatase method after antigen retrieval. Staining with mAb T311 showed a sensitivity of 94% for melanoma with a very high specificity for melanocytic cells. Immunopositivity (94% of melanomas overall) correlated inversely with clinical stage: clinical stage I and stage II showed 100%, stage III and stage IV 86% immunoreactivity each. Staining changed from an exclusively homogeneous pattern in early stages to a more heterogeneous pattern in later stages. Melanocytic control tissue like nevi of different subtypes all showed weak to moderate, homogeneous immunoreactivity with polarity towards the epidermis. RT-PCR ELISA analysis of short-term melanoma cell cultures displayed mRNA expression in only half of the originally immunopositive tumors only, suggesting rapid mRNA expression loss in culture. mAb T311 allows detection of melanoma-associated tyrosinase protein expression and thus profiling of melanomas using routine archival tissue suited for immunotherapy approaches involving tyrosinase derived epitopes.
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  • 3
    ISSN: 1432-069X
    Keywords: Key words Melanoma ; RT-PCR ELISA ; Melanoma-associated antigens ; Tyrosinase ; gp100 ; Melan A/MART-1 ; MAGE-3 ; MUC18
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Fresenius' journal of analytical chemistry 360 (1998), S. 426-429 
    ISSN: 1432-1130
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Open-path monitoring of the atmosphere using Fourier transform infrared spectrometry has recently become a useful real-time in situ analytical technique. The U.S. Environmental Protection Agency (EPA) is currently formulating Method TO-16, a protocol for infrared remote sensing of the hazardous air pollutants (HAPs) identified in EPA’s Clean Air Act of 1990. To support infrared based sensing technologies, the National Institute of Standards and Technology (NIST) is currently developing a standard quantitative spectral database of the HAPs based on gravimetrically prepared samples. This paper presents the protocol used to prepare the gravimetric mixtures and initial results.
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  • 5
    ISSN: 1432-2048
    Keywords: Auxin-binding protein ; Glycine max L ; Guanosine nucleotides ; Phospholipase A2 ; Signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The molecular mechanism of membrane-associated reactions induced by auxin was investigated in membranes isolated from cultured cells of soybean (Glycine max L.). Auxins increased the activity of phospholipase A2 in microsomes isolated from suspensioncultured soybean cells. The reaction was measured as the accumulation of radioactive lysophosphatidylcholine hydrolyzed from radioactive phosphatidylcholine in membranes which had been prelabelled with [14-C]choline in vivo. Stimulation by auxin was detectable after 1 min and was auxin-specific in that weak auxins had little effect. Auxin concentrations as low as 2·10−8 M and up to 2·10+3 M α-naphthaleneacetic acid already stimulated the phospholipase A2 activity. Guanosine and adenosine diphosphate at 100 μM, if applied during homogenization of cells, completely abolished the stimulation of phospholipase A2 by auxin and, when applied after homogenization, had no effect. Guanosine and adenosine 5′-O-thiotriphosphate, uridine 5′-diphosphate, and uridine 5′-triphosphate, all at 100 μM, had no effect in either treatment, suggesting that only nucleotides entrapped in the vesicles could exert an effect. The effect of auxin on phospholipase A2 had an optimum at pH 5.5 and was abolished completely by an antibody against the membrane-associated auxin-binding protein from maize coleoptiles, applied after homogenization. This antibody recognized a 22-kDa polypeptide in highly purified plasma membranes from cultured soybean cells. This suggests a receptor function for this auxin-binding protein and a role for a cytosolic nucleotide-binding protein in the activation of phospholipase A2 by auxin. It is concluded that phospholipase A2 has a function in plant signal transduction.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-2048
    Keywords: Key words: Auxin ; Auxin inhibitor ; Cucurbita ; Elongation growth ; Phospholipase A inhibitor ; Signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Auxin and elicitors reportedly activate phospolipase A. A number of inhibitors known to inhibit animal phospholipase A2 were tested for their ability to inhibit hormone and fusicoccin-induced growth. To this end, growth induced by indolyl-3-acetic acid and 2,4-dichlorophenoxyacetic acid in hypocotyl segments of etiolated zucchini (Cucurbita pepo L.) seedlings was determined in the presence of the inhibitors nordihydroguajaretic acid (NDGA), aristolochic acid, 5,8,11,14-eicosatetraynoic acid (ETYA), PBx (a prostaglandin derivative), and oleylethyl phosphocholine. Each chemical proved inhibitory to auxin-induced growth, oleylethyl phosphocholine being the least effective. The effects of the first three inhibitors were investigated in more detail. Growth induced by 10 μM 2,4-dichlorophenoxyacetic acid or 1 μM indolyl-3-acetic acid was inhibited 50% by about 30–50 μM NDGA, by about 25 μM aristolochic acid, and by about 10–20 μM EYTA. Growth inhibition was reversible and became apparent 0.5–1 h after inhibitor addition. Growth induced by 0.5 or 1 μM fusicoccin was much less inhibited by NDGA and by ETYA, whereas aristolochic acid was only slightly less effective on fusicoccin-induced than on auxin-induced growth. These three inhibitors were also tested for their effects on gibberellin-induced growth in light-grown peas (Pisum sativum L.) and on cytokinin-induced expansion growth in excised cotyledons from radish (Raphanus sativum L.) seedlings. In both tests, aristolochic acid had toxic side-effects although gibberellin-induced growth was still apparent. In the gibberellin test, neither NDGA at up to 100 μM nor ETYA at 80 μM was inhibitory to hormone-induced growth. Moreover, 40 μM ETYA was not inhibitory to kinetin-induced growth. We hypothesize that the selectivity of phospholipase A2 inhibitors for auxin-induced growth implies a different signal transduction pathway for each of the different signal substances tested, and that auxins might use fatty acid(s) and/or lysophospholipid(s) or their derivatives as the preferred second messengers.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Planta 203 (1997), S. S107 
    ISSN: 1432-2048
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-2048
    Keywords: Glycine ; Vacuolar H−+-ATP-ase ; Tonoplast (negative staining)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Highly purified tonoplast fractions isolated by preparative free-flow electrophoresis from hypocotyls of etiolated soybean (Glycine max L. (Merr.)) were examined by negative-staining electron microscopy, and many but not all vesicles were found to exhibit head and stalk structures resembling the 9-nm stalked F1 ATPase particles reported previously for Neurospora (Bowman et al., 1989, J. Biol. Chem. 264, 15606–15612). The structures show distinguishing characteristics similar to those for Neurospora. These include a cleft in the particle not exhibited by mitochondrial F1 ATPase and a tendency to disappear from the membrane when treated with nitrate plus Mg−2+-ATP-containing solutions. The position of the stalked ATPase structures, indicates that some of the tonoplast vesicles were oriented cytoplasmic side out whereas others were oriented cytoplasmic side in.
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  • 9
    ISSN: 1432-2048
    Keywords: Auxin receptor ; Cucurbita (auxin action) ; Growth ; Phospholipase A in auxin action ; Signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Microsomal vesicles were prepared from zucchini (Cucurbita pepo L.) hypocotyls containing radioactive phosphatidylethanolamine or phosphatidylcholine, and these lipids were used as substrates by phospholipase A which is activated by auxins. Phospholipase D and phospholipase C hydrolysed the same substrates but were not influenced by auxin. Phospholipase A was activated by the auxins indolyl-3-acetic acid, 2,4-dichlorophenoxyacetic acid and, to a lesser extent, by α-naphthaleneacetic acid whereas the weak auxins 2,3-dichlorophenoxyacetic acid and β-naphthaleneacetic acid were almost inactive. This hormone specificity was also found in growth tests with etiolated zucchini hypocotyls. Phospholipase A activation by auxin was blocked by a polyclonal antibody against the maize auxin-binding protein. We propose that phospholipase A activation is a primary reaction in the signal transduction leading from hormone-binding to the growth response.
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  • 10
    ISSN: 1432-2048
    Keywords: Cucurbita ; Filipin ; H+-ATPase ; Lepidium ; Plasma membrane ; Tonoplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Large-scale preparations of highly purified tonoplast and plasma-membrane vesicles were obtained from roots (garden cress, Lepidium sativum L.) and shoots (etiolated zucchini hypocotyl, Cucurbita pepo L.) of representative dicotyledonous seedlings. When tonoplast-enriched fractions of cress roots were prepared by centrifugation and then subjected to free-flow electrophoresis a highly purified tonoplast fraction was obtained. This fraction from cress roots was characterized by morphometry of filipin-treated freeze-fractured preparations and by enzymology to be about 90% homogeneous. Using latency of nitrate-inhibited ATPase and H+-pumping as criteria we found that the majority of the tonoplast vesicles from both sources were oriented right(cytoplasmic)-side-out. Plasma-membrane vesicles were first purified by two-phase partitioning and then subjected to free-flow electrophoresis for further purification. From cress roots, the fraction of highest purity contained 89% plasma-membrane vesicles as judged by morphometry of filipin-treated, freeze-fractured preparations and by enzymology. From both sources, the major plasma-membrane subfraction in the upper phase after two-phase partitioning was shown to have the least electrophoretic mobility in free-flow electrophoresis and to be oriented right(extracytoplasmic)-side-out a slightly more mobile plasma-membrane subfraction was oriented inside-out and originated after freezing thawing from outside-out plasma-membrane vesicles.
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