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  • Analytical Chemistry and Spectroscopy  (9)
  • Cell & Developmental Biology  (6)
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  • 1
    ISSN: 0886-1544
    Keywords: cytoskeletal maturation ; keratinlike filaments ; holocrine secretion ; cytoskeleton ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Previous studies with the hagfish, a primitive vertebrate, have shown that the gland thread cells (GTCs) each contain a single thread (∼60 cm long in average-sized cells) in the form of a concisely coiled cytoskeletal entity destined for export by holocrine secretion. The thread in relatively immature GTCs consists almost entirely of intermediate filaments (IFs) bundled in parallel alignment with far fewer microtubules (MTs). The three thread polypeptides described earlier (α, basic; β acidic; γ, most acidic; each with a Mr of 63-64 kD) are now further evaluated with respect to in vitro assembly, cross-reactivity with IF polypeptides from higher vertebrates, and peptide sequence homology with known IF polypeptides. The overall results mainly suggest that the hagfish polypeptides are keratinlike substances but lamins or a new type of IF is not ruled out. However, cross-reactivity is weak with mammalian keratins; the 8-11-nm filaments formed from mixtures of α and γ in vitro are generally linear rather than the curvilinear structures usually formed by keratin and nonkeratin IFs; and mixtures of α and β tend to yield 9-12-nm granules or granular strings. Polypeptide analyses on GTCs segregated on the basis of maturational stage show a progressive increase in β/γ values which correlates with cell maturation, but the α/(β+γ) ratios remain near 1. Inasmuch as β and γ have many similar properties, the documented increase in the amount of the β component in aging GTCs might in part be the result of a failure in a posttranslational modification system and may contribute to the ultrastructural changes that accompany thread maturation in preparation for holocrine secretion and subsequent modulation of the viscoelastic properties of mucus.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0935-6304
    Keywords: Crosslinked stationary phases ; Open-tubular capillary liquid chromatography column ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Crosslinked polysiloxane stationary phases were prepared on soda-lime glass capillaries and applied to the separation of polynuclear aromatic hydrocarbons and of phthalates in reversedphase liquid chromatography. Preparation procedures and chromatographic performance of these columns are described.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 0935-6304
    Keywords: Epoxytriacylglycerols ; high temperature gas chromatography ; hydroxytriacylglycerols ; ketotriacylglycerols ; lipids ; negative chemical ionization mass spectrometry ; SOP-50 ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: ---Five seed oils consisting of unusual triacylglycerols have been examined by high temperature gas chromatography using glass capillary columns coated with the stationary phase SOP-50 (50%-diphenyl-50%-dimethylpolysiloxane, methoxy-terminated). The investigated hydroxy, epoxy, and ketokonjuene triacylglycerols were first derivatized in order to permit analysis by high temperature gas chromatography. Structural elucidation of the intact triacylglycerols was performed with high temperature gas chromatography/negative chemical ionization-mass spectrometry using NH3 as a reagent gas. Individual derivatized lipid species with a molecular weight up to 1176 g/mol could be identified.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 0935-6304
    Keywords: cis-13-Eicosenoic Acid (Paullinic Acid) ; Cyanolipids ; Fatty acids ; GLC-MS ; NMR spectroscopy ; Oxazoline derivatives ; Paullinia elegans Sapindaceae ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The fatty acids of the total lipids, triglycerides, and the cyanolipid of Paullinia elegans (Sapindaceae) have been analyzed as their methyl esters and 4,4-dimethyloxazoline derivatives by gas chromatography-mass spectrometry. It was shown that the gas chromatographic separation of the oxazoline derivatives was sufficient to ensure the correct identification of the monoenic fatty acid positional isomers. Stereochemistry of the double bonds has been confirmed by infrared and 13C-nuclear magnetic resonance (NMR) spectroscopy. cis-13-Eicosenoic Acid (Paullinic acid) (44.4%) and cis-11-octadecenoic acid (cis-vaccenic acid) (19.8%) were found to be the main components beside other monoenoic acid positional isomers. The cyanolipid of P. elegans was identified as 2,4-dihydroxy-3-methylenebutyronitrile derivative by IR, 1H-NMR, 13C-NMR spectroscopy and was quantified by 1H-NMR spectroscopy (71.4%).
    Additional Material: 3 Ill.
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  • 5
    ISSN: 0935-6304
    Keywords: α-Monoacetotriacylglycerides ; Celastraceae ; Fatty acids ; GLC ; GLC-(NCI)MS ; Maytenus seed oils ; OV-225-OH, OH-terminated ; SOP-50, methoxy-terminated ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The seed oils of three Maytenus species have been examined by high temperature GLC using SOP-50 (methoxy-terminated) and OV-225-OH (OH-terminated) glasscapillary columns. The main compounds (71-76%) could be identified as α-mono-acetotriacylglycerols by a combination of high temperature GLC-MS in negative chemical ionization mode with a CH4/N2O mixture as reactant gas and other spectroscopical methods (infrared, 1H- and 13C-nuclear magnetic resonance spectroscopy). The OV-225-OH phase demonstrated better separation properties that the SOP-50 phase for this substance class and up to eight species of α-monoacetotriacylglycerols could be separated. The content of usual triacyclglycerols was below 1%. The results of the analysis of the individual fatty acids as their methyl esters by GC and GC-MS are also presented.
    Additional Material: 2 Ill.
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  • 6
    ISSN: 0749-1581
    Keywords: Phase-editing ; Relayed responses ; 2D HMQC-TOCSY ; NMR Spectra ; Digoxin ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: An NMR technique for the determination of proton vicinal couplings relayed at the carbon chemical shift is presented. With this technique, which is an extension of DEPT-HMQC-TOCSY, it is possible to generate phaseedited 2D NMR data in which the 1H—13C relayed responses differ in phase as a function of the proton multiplicity of the carbon from whose proton(s) the response originates. A simple pulse scheme is presented to allow further simplification of the spectrum by eliminating the direct 1H—13C correlations. Thus, responses arising from methines or methyls will be of opposite phase from responses originating from methylenes; direct correlations are eliminated to minimize congestion. An example is presented with the cardiac drug digoxin.
    Additional Material: 4 Ill.
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  • 7
    ISSN: 0749-1581
    Keywords: NMR ; Region-selective inverse detected long-range heteronuclear chemical shift correlation ; Shaped pulses ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: F1 region-selected variants of the HMBC experiment were compared and evaluated. In particular, a significant advantage was demonstrated for an E-BURP-2 pulse over a Gaussian 270° pulse for region-selected applications. The Gaussian 270° pulse would still, however, be the preferred pulse for use in single-resonance, on-resonance selective excitation. Data are presented from NMR spectrometers both equipped and not equipped with programmable pulse modulators (waveform generators). Pulse evaluations were performed using the simple heteroaromatic alkaloid quindoline; application of the technique to the important immunosuppressive cyclic peptide cyclosporin-A is also presented and discussed.
    Additional Material: 9 Ill.
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  • 8
    ISSN: 0730-2312
    Keywords: EGF derivative ; EGF receptor ; cytochemical detection ; clinical oncology ; tumor marker ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: A biotinylated derivative of murine epidermal growth factor (EGF) was prepared by covalent attachment of the terminal amino group of EGF to N-biotinyl-ε-aminocaproyl-N-hydroxysuccinimide. The stoichiometry of biotin incorporation was in the range of one biotin moiety per EGF molecule. The biotinylated EGF (biotinyl-ε-caproyl-EGF, BioEGF) binds to EGF receptors on intact Ehrlich ascites carcinoma (EAC) cells with an affinity similar to that of native EGF and displays the same mitogenic activity as EGF in a soft agar test system with normal rat kidney (NRK) cells. BioEGF was visualized on cultured cells and tissue sections of a head and neck tumour by commercial streptavidin/avidin detection systems. Cytochemical analyses of certain tumour forms can be easily performed using the BioEGF probe.
    Additional Material: 6 Ill.
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  • 9
    ISSN: 0730-2312
    Keywords: epidermal growth factor receptor ; breast cancer ; immunohistochemistry ; cell proliferation ; receptor phosphorylation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The prognostic significance of the epidermal growth factor receptor status (EGF-R-status) for certain human tumors requires the development of antibodies useful for clinical application. We used purified receptor preparations to generate monoclonal antibodies immunoreactive with the EGF-R purified from placenta membranes and A431 tumors. Four of the hybridomas contained antibodies (R2, R3, R5, and R9) which recognized both antigens. Antibody R3 was shown to display the following properties: it binds with a KD value of about 10-9-10-10 M to the receptor, a half maximal inhibition of EGF-binding is achieved at 5 × 10-8 M, and in Western blots of cell membranes R3 specifically detects the EGF-R at 0.1 μ/ml. R3 inhibits EGF-dependent clonogenic growth of NRK cells and completely blocks EGF stimulated autophosphorylation of the receptor. Moreover, R3 also detects EGF-R in paraffin-embedded tissue sections taken from human salivary gland, term placenta, and adult skin and mammary carcinomas. Thus, R3 can be used in retrospective diagnostic clinical studies and might help to develop new immunotherapeutic intervention.
    Additional Material: 6 Ill.
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  • 10
    ISSN: 0730-2312
    Keywords: explant culture ; stimulation of DNA synthesis ; inhibition of functional differentiation ; endogenous TGFα ; arachidonic acid release ; phospholipase A2 ; metabolic inhibitors ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Epidermal growth factor (EGF) has been suggested to be involved in mammary gland development by mitogenic stimulation of the ductal and alveolar epithelium in virgin mice. The present studies demonstrate that also in late-pregnant mice EGF leads to proliferation of the ductal, ductular, and alveolar epithelium. The mitogenic effect is associated with structural and functional dedifferentiation of alveolar cells as revealed by analysis of morphology, expression of cytosolic and secretory proteins, and fatty acid synthesis. Using a combination of metabolic inhibitors, the dedifferentiating effect of EGF could be blocked while the mitogenic action was not influenced. This finding demonstrates that the signal transduction pathway leading to dedifferentiation and mitosis can be separated, and that the dedifferentiating effect of EGF is independent of its mitogenic properties, but is probably mediated by activation of the arachidonic acid-dependent pathways (cyclo- and lipoxygenase pathways). Release of arachidonic acid from the endogenous phospholipid pool was found to be an early response of the explants to EGF. Accordingly, arachidonic acid itself proved to be capable of inducing epithelial dedifferentiation but failed to stimulate proliferation. TGFα showed qualitatively similar effects as EGF but was generally a stronger agonist. It is suggested that EGF and TGFα also play a role in mammary gland physiology during pregnancy by final developing and maintanance of the lobulo-alveolar structure in the mammary gland and prevention of premature onset of lactation, and that this is mediated through the PLA2-arachidonic acid signalling cascade.
    Additional Material: 4 Ill.
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