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  • 1
    ISSN: 0730-2312
    Keywords: β1→3N-acetylglucosaminyltransferase ; cancer-associated carbohydrate antigens ; biosynthesis ; glycosphingolipid ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Human colonic adenocarcinoma DLD-1 cells were grown under conditions which induce characteristics of differentiated cells using medium containing 0.8% N,N-dimethylformamide in order to study alterations in glycosphingolipid glycosyltransferase activities during this process. Analysis of biosynthetic reactions involved in lacto-series antigen synthesis revealed no changes in the specific activities of either β1→4galactosyltransferase or α1→3/4fucosyltransferase with N,N-dimethylformamide treatment. However, a dramatic decrease of from 14- to 20-fold in the β1→3N-acetylglucosaminyltransferase activity was observed in the treated cells. This enzyme catalyzes the rate-limiting step in lacto-series core chain synthesis. This is consistent with the pattern of regulation of lacto-series antigen expression found to occur during oncogenesis in human colonic mucosa (Holmes EH, Hakomori S, Ostrander GK: J Biol Chem 262:15649, 1987). Total glycolipids from untreated and N,N-dimethylformamide-treated cells were isolated and subjected to TLC immunostain analysis and solid phase radioimmunoassay with a series of monoclonal antibodies specific for lacto-series-based carbohydrate antigens. A decrease of about 2-fold or less in the quantity of lacto-series antigens was observed as a consequence of N,N-dimethylformamide treatment in both neutral glycolipid and ganglioside fractions. The results suggest that only very low levels of β1→3N-acetylglucosaminyltransferase activity are required for the steady state expression of significant levels of lacto-series based glycolipids and that modulation of its activity levels by N,N-dimethylformamide treatment in DLD-1 cells represents a convenient in vitro system for studying aspects of regulation of lacto-series antigen expression.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0730-2312
    Keywords: IGF-I receptor ; T-cells ; OKT-3 ; PHA ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The biological effects of the IGFs are mediated through interaction with specific cell surface receptors. It has been previously reported that mitogenic activation of T-lymphocytes by phytohemagglutinin (PHA) is associated with increased IGF-I receptor content. However, the mechanisms which regulate IGF-I receptor expression during T-lymphocyte activation are unknown. To explore further the regulation of IGF-I receptor expression in T-cells, we investigated IGF-I receptor content and mRNA abundance in T-lymphocytes after stimulation either by PHA or OKT-3, the latter being a monoclonal antibody directed against the CD-3 antigen of the T-cell receptor IGF-I binding in T-cells demonstrated increased IGF-I receptor content after stimulation by both PHA and OKT-3. Peak binding was induced after 72 h of treatment with PHA and 48 h of treatment with OKT-3. Affinity cross-linking of 125I-IGF-I to T-cell membranes demonstrated a single ∼ 130 kDa band which was increased after treatment with PHA or OKT-3. This band was inhibited by the addition of α-IR3, a monoclonal antibody to the IGF-I receptor. Both PHA and OKT-3 increased IGF-I receptor mRNA abundance with peak increases at 20 h and 60 h, respectively. Parallel increases in IGF-I receptor and β-actin mRNA abundance were observed, consistent with previous studies demonstrating increased actin gene expression after T-cell activation. Thus, the increase in IGF-I receptor mRNA abundance markedly preceded the increase in IGF-I receptor content after PHA stimulation, but the increase in IGF-I receptor mRNA abundance followed the increase in IGF-I receptor content after OKT-3. These studies suggest, therefore, that IGF-I receptor content in both of these activated cells is not regulated primarily at the level of steady state mRNA.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 50 (1992), S. 55-62 
    ISSN: 0730-2312
    Keywords: aberrant crypts ; chemoprevention ; enzyme-altered foci ; intermediate biomarker ; preneoplastic lesions ; putative colorectal cancer precursor ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Aberrant crypts are recognized in methylene blue-stained, unsectioned, colonic mucosa by their increased size, elliptical lumenal opening, thicker epithelial layer, and increased pericryptal region. Aberrant crypt foci in rodents are observed as early as 2 weeks and for at least 9 months after a single dose of carcinogen, have a distribution that parallels that of tumors, and have an increased number of aberrant crypts per focus with time after the carcinogen dose. The ability to quantify these lesions in the entire colon of rodents in less than an hour suggests that aberrant crypts may provide a highly efficient in vivo bioassay for colon carcinogens. Since aberrant crypt foci appear to be the earliest identifiable putative precursors of colon cancer, they represent lesions that can be characterized further for the earliest genetic and biochemical alterations. In F344 rats, we have demonstrated that aberrant crypts have multiple histochemically-detectable enzyme alterations. Using similar techniques, we were the first to demonstrate aberrant crypts in unsectioned human mucosa. After embedding and sectioning, these microscopic aberrant crypts resemble rare lesions described earlier in the literature after extensive serial sectioning. In rats and humans, aberrant crypts may be histologically normal or display varying degrees of dysplasia and histochemically-detectable altered enzyme activities. These putative, preneoplastic lesions should reveal early changes that precede colon cancer and ways to alter their progression.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 0730-2312
    Keywords: β1 → 3galactosyltransferase ; stable expression ; glycolipids ; lacto-series type 1 chain ; Lewis antigens ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Transient expression of a human colonic adenocarcinoma Colo 205 cell derived cDNA in cell lines which ordinarily express only neolacto-series glycolipids has resulted in the expression of a β1 → 3galactosyltransferase gene responsible for synthesis of glycolipids based upon the lacto-series type 1 core chain. Calcium phosphate transfected cells were panned on anti-lgM coated plates after initial treatment with a combination of monoclonal antibodies specific for type 1 chain terminal structures (TE-3) and a very broadly specific antibody reactive with multiple type 1 chain derivatives (TE-2). Adherent cells after panning were capable of efficiently transferring Gal in β1 → 3-linkage to the acceptor glycolipid Lc3. Using these reagents, clones of stably transfected human colonic adenocarcinoma HCT-15 cells were produced and isolated. Parental HCT-15 cells do not express type 1 chain based antigens. The nature of the type 1 chain based antigens produced in each of these clones was analyzed by solid phase antibody binding assays. Three types of behavior were observed. Formation of type 1 terminal structures that were either exclusively sialylated or fucosylated, or a mixture of sialylated and fucosylated determinants occurred. In contrast, no difference in type 2 antigen expression between any clone and the parental cells was observed. These data suggest that coordination of subsequent reactions capable of modifying type 1 chain structures is not the same in all clones. The relationship of these results to aspects of cellular regulation of carbohydrate biosynthesis is discussed. © 1992 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 33 (1992), S. 436-442 
    ISSN: 1040-452X
    Keywords: Transgenic animals ; DNA methylation ; Concatemer formation ; Mosaicism ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The state of genes microinjected into mouse embryos was followed from the one-cell to the blastocyst stage using the polymerase chain reaction (PCR). Microinjected DNA was detected in all one-, two-, and four-cell injected embryos and in 44% of morula and 26% of blastocysts. Head-to-tail ligation of microinjected genes, a common feature of stably integrated transgene arrays, was detected in all embryos after injection of microinjected genes and occurred irrespective of the structure at the ends of the injected genes. Sensitivity of microinjected DNA to a methylation-dependent restriction endonuclease Dpn I was lost in all embryos by the two-cell stage (24 hr), indicating a change in DNA methylation, independent of transgene integration. Dissociation of blastomeres prior to compaction revealed a mosaic distribution of the microinjected DNA within the embryo and supports the notion that injected genes form a limited number of arrays, which segregate independently until they integrate into the genome or are degraded. Published 1992 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Bioelectromagnetics 12 (1991), S. 319-333 
    ISSN: 0197-8462
    Keywords: pulse-train ; dispersive dielectrics ; Fourier transform ; focusing ; hot-spot ; Life and Medical Sciences ; Occupational Health and Environmental Toxicology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Physics
    Notes: In analytical studies, we investigated induced-field patterns and SAR distributions in a lossy, dispersive, homogeneous, dielectric sphere typical of muscle tissue as irradiated by a plane-wave pulse train consisting of a pulse-modulated sinusoidal carrier wave. Calculations were made for carrier frequencies of 1, 3, and 15 GHz, pulse widths of 0.333, 2.0 and 4 ns, and pulse repetition rates of 1.11 × 106, 100 × 106, and 181.18 × 106 pps. The classical Mie solution was modified for a train of incident pulses that was represented by a Fourier series, and the fast-Fourier transform was used to sum the series. Computationally, the technique proved to be feasible and less expensive than we expected. The calculated field patterns show that the sphere's physical dimensions and the internal wavelength of the carrier greatly influence the nature of pulse-train propagation in the sphere. Harmonics having internal wavelengths nearly equal to the radius of the sphere produce most of the absorption; other harmonics produce little absorption. An intense hot spot is observed in spheres with radii that match the carriers' wavelengths.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In previous experiments we have shown that acute (30 minutes) exposure to phorbol esters or other protein kinase C activators causes increased transepithelial permeability, specifically by the increased paracellular permeability through tight junctions. However, the role of protein kinase C activators in carcinogenesis is predicted upon a chronic exposure of an effective dose at frequent intervals for a prolonged period of time. We therefore sought to determine the effect of chronic phorbol ester exposure on transepithelial permeability by exposing cells of the polar renal epithelial cell line, LLC-PK1, to phorbol esters for time periods as long as 16 weeks. The following changes ensued: (1) after the initial drop in transepithelial resistance due to phorbol ester exposure, i.e., an increase in transepithelial permeability (in the acute phase of exposure), an adaptive response occurs as transepithelial resistances in chronically exposed cultures recover to approximately 50% of control values, (2) the cell sheets in chronically exposed cultures lose their acute responsiveness of transepithelial permeability to phorbol ester exposure, (3) cell sheet architecture changes as cells occasionally multilayer and actual polyp-like cell masses appear at high frequency, and (4) cytosolic protein kinase C activity decreases to 50% of control level with acute exposure and then is further decreased to less than 1% of control level in chronically treated cells; membrane-associated PKC activity is not as sharply decreased. The possible role of transepithelial permeability in carcinogenesis and the value of chronically treated epithelial cell cultures as a model for two-stage carcinogenesis are discussed. © 1992 Wiley-Liss, Inc.
    Additional Material: 9 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 145 (1990), S. 531-535 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The ability of 12-0-tetradecanoylphorbol-13-acetate (TPA) to stimulate mitogenesis in BALB/c-3T3 cells and in a Na+K+C1- -cotransport-defective variant subclone was investigated. This transport variant had previously been reported to be TPA mitogenically nonresponsive (O'Brien and Prettyman: Journal of Cellular Physiology 130:377-381, 1987) since the addition of TPA to the spent medium of density-arrested cultures stimulated DNA synthesis in the parent but not the variant cell line. We now report that the addition of TPA plus insulin, either directly to the spent medium or together with fresh medium, stimulated DNA synthesis in both the parent and variant cells to approximately the same extent. The parent and transport-defective cells differed, however, in their sensitivity to the co-mitogenic effects of insulin or insulin-like growth factors.
    Additional Material: 3 Ill.
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  • 9
    ISSN: 0044-2313
    Keywords: Dysprosiumdiiodide ; dysprosium triiodide ; enthalpy ; electrode potential ; Chemistry ; Inorganic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Description / Table of Contents: Bestimmung der Bildungsenthalpien von DyI2 und DyI3 sowie Abschätzung des Dy3+/Dy2+-Standardelektrodenpotentials in wäßrigem MediumDyI2 and DyI3 wurden nach Literaturmethoden hergestellt. Ihre Lösungsenthalpien wurden bestimmt und die Bildungsenthalpien zu ΔfH°(DyI3, s, 298 K) = -(394 ± 16) kJ · mol-1 und ΔfH°(DyI2, s, 298 K) = -(616 ± 10) kJ· mol-1 berechnet. Entsprechenden Literaturangaben sowie geschätzten Lösungsenthalpien und Standardentropien wurde E°(Dy3+/Dy2+, aq.) zu -(2.6 ± 0.2) V berechnet. Ein Vergleich der Enthalpien für die Reduktion von DyI3 zu DyI2 bzw. DyI zu DyI2 wird vorgenommen.
    Notes: DyI2 and Dy3I were synthesized by literature techniques. Their enthalpies of solution were determined and their enthalpies of formation calculated to be ΔfH°(DyI2, s, 298 K) = -(394 ± 16) kJ· mol-1 and ΔfH°(DyI3, s, 298 K) = -(616 ± 10) kJ· mol-1. With appropriate literature and estimated enthalpies of solution and standard entropies, the E°(Dy3+/Dy2+, aq) was calculated to be -(2.6 ± 0.2) V. A comparison is made of the enthalpies of reduction of DyI3 to DyI2 and of DyCl3 to DyCl2.
    Additional Material: 2 Tab.
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  • 10
    ISSN: 0749-503X
    Keywords: Superoxide dismutase ; cytochrome P450 ; chromosome VIII ; Saccharomyces cerevisiae ; polymorphisms ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: DNA sequencing and analysis of genomic DNA using the polymerase chain reaction were used to demonstrate that SOD1 and ERG11 are adjacent genes in Saccharomyces cerevisiae S288c and to establish the correct intergenic sequence of this segment on chromosome VIII.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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