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  • 1
    ISSN: 1600-0625
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Notizen: Abstract:  Protein kinase C (PKC) isoforms play pivotal roles in the regulation of differentiation of normal human epidermal keratinocytes (NHEK). In this study, we investigated the participation of the PKC system in the proliferation and high cell density-induced differentiation of the human immortalized keratinocyte line HaCaT. HaCaT keratinocytes possessed a characteristic PKC isoform pattern (PKCα, β, γ,δ,ε,η,θ,ζ), which altered during proliferation and differentiation. The GF109203X compound, a selective PKC inhibitor, suppressed the expressions of the late (granular cell) differentiation markers involucrin (INV) and filaggrin (FIL), and the terminal marker keratinocytes-specific transglutaminase-1 (TG), but did not affect the level of the early (spinous cell) marker keratin 10 (K10) and cellular proliferation. Phorbol 12-myristate 13-acetate (PMA), an activator of PKC, inhibited proliferation, elevated intracellular calcium concentration, decreased the expression of K10, and increased the expressions of INV, FIL, and TG. These data indicate that the endogenous activation of PKC regulates the expressions of the late differentiation markers, and that the exogenous activation of PKC by PMA results in the induction of terminal differentiation. Because the cellular effects of PMA were accompanied by differential down-regulations of the sensitive PKC isoforms in proliferating and differentiating cultures, our findings argue for the differential roles of the existing PKC isoforms in the regulation of cellular proliferation and high cell density-induced differentiation of HaCaT cells.
    Materialart: Digitale Medien
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  • 2
    Digitale Medien
    Digitale Medien
    Oxford, UK : Munksgaard International Publishers
    Experimental dermatology 11 (2002), S. 0 
    ISSN: 1600-0625
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Notizen: Abstract: Capacitive calcium influx is associated with the release of calcium from internal stores and participates in intracellular calcium homeostasis. In keratinocytes, its activation is linked to the stimulation of the phospho-inositide (PI) pathway and seems to be altered in psoriasis. An overnight treatment of isolated HaCaT keratinocytes with phorbol 12-myristate 13-acetate (PMA) selectively downregulated the classical, calcium-dependent protein kinase C (PKC) isoenzyme PKCα in preconfluent cells. This was parallelled by an increased capacitative calcium influx with no effects on the PI pathway. These observations were strengthened in measurements using cyclopiazonic acid which revealed a 47% increase in PMA pretreated as compared with control cells in the calcium influx rate through store-operated calcium channels (SOC-s) following the emptying of the intracellular calcium stores. In confluent as compared with preconfluent cultures PKCε was markedly increased, while other isoenzymes were not affected. In parallel, the kinetics of capacitative calcium influx were altered, showing clear inactivation. PMA pretreatment in these cells had little effect on PKCα but downregulated both PKCβ and PKCε, and did not increase the influx through SOC-s. These observations support the differential regulation of SOC-s by PKC and suggest the involvement of several PKC isoenzymes in human keratinocytes.
    Materialart: Digitale Medien
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  • 3
    Digitale Medien
    Digitale Medien
    Osney Mead, Oxford OX2 0EL, UK : Blackwell Scientific Publication
    Molecular microbiology 17 (1995), S. 0 
    ISSN: 1365-2958
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie , Medizin
    Notizen: Rhizobium fredii is a nitrogen-fixing bacterial symbiont of soybean and a number of other legume species. We have studied the transcriptional organization of a Sym plasmid locus that restricts the host range of R. fredii USDA257 at both the host species and cultivar level. The genes of this host-specificity locus, noIXWBTUV, are transcribed from three promoters. Two of these, which are upstream of noIW and noIBTUV, are oriented face to face and initiate transcription at sites that are 14 bp apart. The third lies upstream from noIX. The noIW promoter is constitutive, whereas the noIB and noIX promoters are inducible by flavonoid signals. We have attempted to express genes from this locus in Escherichia coli systems, both in vivo and in vitro. We detected the insert and orientation-specific expression of two genes, noIX and nolW, but we were unable to obtain expression of noIBTUV. Antiserum raised against NoIT nevertheless detected an abundantly expressed polypeptide of the predicted size in protein extracts of USDA257. This observation, as well as RNA dot blot data from a series of mutants, indicates that noIBTUV is expressed as a single transcriptional unit in R. fredii. Immunological detection of NoIT, and of a second protein, NoIX, was strictly dependent on flavonoid induction. The NoIX protein was larger than the size predicted from the previously published nucleotide sequence, and this led to resequencing and revision of the open reading frame.
    Materialart: Digitale Medien
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  • 4
    ISSN: 1546-170X
    Quelle: Nature Archives 1869 - 2009
    Thema: Biologie , Medizin
    Notizen: [Auszug] The urinary tract functions in close proximity to the outside environment, yet must remain free of microbial colonization to avoid disease. The mechanisms for establishing an antimicrobial barrier in this area are not completely understood. Here, we describe the production and function of the ...
    Materialart: Digitale Medien
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  • 5
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Publishing Ltd
    British journal of dermatology 104 (1981), S. 0 
    ISSN: 1365-2133
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Medizin
    Notizen: A vesiculobullous eruption with clinical and histological features of bullous pemphigoid developed in a 38-year-old woman with proved systemic lupus erythematosus. The patient had a sulphone responsive blistering disease that was characterized by pruritic subepidermal bullae and linear, predominantly IgA basement membrane zone deposition and IgA pemphigoid antibodies in her sera. Because both diseases are associated with immune complexes of special immunoglobulin classes, this association may not be entirely fortuitous.
    Materialart: Digitale Medien
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  • 6
    ISSN: 1432-0533
    Schlagwort(e): Key words Skeletal muscle ; Human ; Differentiation ; Protein kinase C ; Isozymes
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract The mechanism of skeletal muscle regeneration in vivo can be well modeled in vitro by culturing skeletal muscle cells. In these cultures mononuclear satellite cells fuse to form polynuclear myotubes by proliferation and differentiation. The aim of this study was to determine how the different protein kinase C (PKC) isozymes were expressed during differentiation of human skeletal muscle in vitro. The expressions of desmin, used as a muscle-specific intermediate filament protein marker of differentiation, and of different PKC isozymes were detected by single and double immunohistochemical labeling, and by Western blot analysis. In skeletal muscle cells we could identify five PKC isozymes (PKCα, -γ, -η, -θ and -ζ). The expressions of PKCα and -ζ did not change significantly during differentiation; their levels of expression were high in the early immature cells and remained unchanged in later phases. In contrast, the expression levels of PKCγ and -η increased with differentiation. Furthermore, the cellular localization of PKCγ markedly altered during differentiation, with a perinuclear-nuclear to cytoplasmic translocation. The change in the level of expression of PKCθ during differentiation showed different pattern; its expression was high during the early phases, but a decreased immunostaining was detected in the matured, well-differentiated myotubes. We conclude, therefore, that cultured human skeletal muscle cells possess a characteristic PKC isozyme pattern, and that the different phases of differentiation are accompanied by different expression patterns of the various isozymes. These data suggest the possible functional and differential roles of PKC isozymes in human skeletal muscle differentiation.
    Materialart: Digitale Medien
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  • 7
    ISSN: 1432-069X
    Schlagwort(e): Key words Keratinocytes ; Intracellular calcium ; Signal transduction ; Heterogeneity
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Intracellular calcium release induced by transient applications of phosphoinositide agonists was measured using adherent single HaCaT keratinocytes loaded with the acetoxymethyl derivative of fura-2. Application of ATP, bradykinin and formyl-Met-Leu-Phe (fMLP) resulted in a transient increase in intracellular calcium concentration ([Ca2+]i) with an average half-width of 40 ± 21 s and a decay time constant of 15 ± 10 s (mean ± SD, n = 108), irrespective of the agonist applied. The cells could be classified into two groups: in 53% of the cells repeated stimulation brought about a progressively smaller change in [Ca2+]i (type 1 cells), whereas in the remaining cells the amplitude of the calcium transients was essentially unchanged (type 2 cells). Furthermore, calcium transients in type 1 cells had broader half-widths and slower decays. No difference was found between the agonists in respect of the characteristics of the evoked calcium transient within each subpopulation. However, bradykinin and fMLP desensitized some cells. These results indicate that the activation of the inositol trisphospate transduction pathway by different agonists induces a characteristic elevation of [Ca2+]i within a given cell. Our results demonstrate that cultured HaCaT keratinocytes are heterogeneous in respect of the calcium transients evoked by the activators of this second messenger system.
    Materialart: Digitale Medien
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  • 8
    ISSN: 1432-2013
    Schlagwort(e): Ca2+ channel blockers Cytoplasmic [Ca2+] transients Fura-2 N-type Ca2+ channels Voltage-gated Ca2+ channels
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract. The significance of voltage-activated Ca2+ currents in eliciting cytoplasmic Ca2+ transients was studied in pyramidal neurones isolated from the rat dorsal cochlear nucleus using combined enzyme treatment/mechanical trituration. Increases in cytoplasmic Ca2+ concentration ([Ca2+]i) were evoked by K+-induced depolarizations (10–50 mM) and monitored by the Fura-2 fluorimetric technique. The acutely dissociated neurones had a resting [Ca2+]i of 17.2±0.5 nM. They possessed caffeine-sensitive Ca2+ stores which were empty at rest; these stores could be filled with Ca2+ entering from the extracellular space and were re-emptied quickly. The effects of various specific high-voltage-activated (HVA) Ca2+ channel antagonists (nifedipine, ω-agatoxin IVA and ω-conotoxin GVIA) on [Ca2+]i transients were tested. Analysis of the blocking effects of these agents on the [Ca2+]i transients indicates that, in the pyramidal neurones of the dorsal cochlear nucleus, N-type Ca2+ channels are primarily responsible for producing the depolarization-induced increases in [Ca2+]i.
    Materialart: Digitale Medien
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  • 9
    Digitale Medien
    Digitale Medien
    Springer
    European journal of nuclear medicine 21 (1994), S. 191-195 
    ISSN: 1619-7089
    Schlagwort(e): Thallium ; Technetium ; Single-photon emission tomography ; Regional cerebral blood flow
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Thallium-201 brain single-photon emission tomography (201Tl-SPET) is widely used to detect viable tumour tissue with increased metabolic activity. When reperfusion takes place early in cerebrovascular lesions of embolic origin, the presence of tissue areas with increased regional blood flow and preserved metabolic activity can also be assumed. In the present study our purpose was to investigate whether or not foci of 201Tl accumulation occur in reperfused areas with sustained morphological integrity indicated by computed tomography (CT) scans not showing hypodensity in the acute or subacute period.In 16 stroke patients with possible cortical embolic infarction, dual 201Tl and technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO) SPET was performed in both the acute and the subacute period. 99mTc-HMPAO SPET was performed to detect reperfusion. Follow-up CT scans from the same period were also available. In five cases 99mTc-HMPAO SPET ruled out reperfusion and 201Tl SPET was also negative. In four cases 99mTc-HMPAO studies indicated reperfusion early in the acute phase (24–72 h), and comparative CT, without showing hypodensity in the acute or subacute period, also favoured the possibility of sustained metabolic activity. In these cases 201Tl SPET was negative in both the acute and the subacute period. In seven cases CT already showed necrosis in 99mTc-HMPAO hypoperfused areas in the acute period, with negative results on corresponding 201Tl SPET. Later reperfusion occurred in the subacute period (8–14 days) as indicated by 99mTc-HMPAO SPET, at which time an unexpected focal accumulation of 201Tl was detected. We cannot give any explanation for the findings, but further studies might clarify the matter and improve our knowledge of the precise mechanism of 201Tl uptake under different conditions. Until then the phenomenon should be borne in mind as a possible pitfall when assessing tissue viability.
    Materialart: Digitale Medien
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  • 10
    Digitale Medien
    Digitale Medien
    Springer
    Molecular genetics and genomics 242 (1994), S. 327-336 
    ISSN: 1617-4623
    Schlagwort(e): Agrobacterium tumefaciens ; Ti plasmid ; Genome mapping ; Plasmid evolution ; Virulence
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Agrobacterium tumefaciens Chry5, a wild-type strain originally isolated from chrysanthemum, is unusually tumorigenic, particularly on soybean. We have mapped the Chry5 Ti plasmid by genomic walking and restriction endonuclease analysis, and have located its virulence, T-DNA, plasmid incompatibility, and l,l-succinamopine utilization loci. Southern analysis has revealed that about 85% of the Chry5 Ti plasmid is highly homologous to another Ti plasmid, pTiBo542. Although all the functions that we have located on pTiChry5 are encoded by pTiBo542-homologous regions, the two Ti plasmids differ in their genetic organization. The overall patterns of restriction sites in the plasmids also differ, with the exception of an approximately 12 kb segment of the virulence region, where the BamHI sites appear to be conserved. Complementation analysis has shown that deletion of a DNA segment which flanks the oncogenic T-DNA results in severe attenuation of virulence. This region also contains a sequence that is repeated in the Chry5 genome outside the Ti plasmid, and that is widely distributed in the Rhizobiaceae.
    Materialart: Digitale Medien
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