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The pattern of expression of the simian virus 40 (SV40) T antigen gene and resultant dysplasia were re-examined in a line of transgenic mice in which the T antigen gene was under the control of the SV40 early promoter. We found that T antigen expression in the kidney, and resulting dysplastic lesions, occurred exclusively in the distal convoluted tubules and the ascending limbs of Henle. Epidermal growth factor (EGF) expression in the kidney of normal mice was similarly immunolocalized. The correlation between high EGF immunoreactivity in normal mouse tissues and T antigen expression in the transgenic counterpart was also seen in the choroid plexus epithelium and in the submandibular glands of male mice. T antigen was not found in the submandibular gland of transgenic females. Similarly, EGF was only rarely detected in the normal female submandibular gland. In contrast to the correlation between T antigen expression in the transgenic mice and EGF expression in the corresponding tissues of the normal mice, within the dysplastic lesions of the transgenic mice EGF expression was severely diminished. Adenocarcinomas of the male submandibular gland from another line of transgenic mice that expresses the Int-1 transgene, showed similarly reduced levels of immunostaining for EGF. Thus, reduced expression of EGF might be a general feature of dysplasia and tumorigenesis in those tissues that normally express EGF. 相似文献
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We examined the ability of opsonized zymosan (OPZ) to stimulate translocation of protein kinase C (PKC) isoforms in human neutrophils. Neutrophils express five PKC isoforms (alpha, betaI, betaII, delta, and zeta), but little is known of their individual roles in neutrophil activation. As determined by immunoblotting, OPZ caused a time-dependent translocation of the predominant PKC isoforms (betaII, delta, and zeta) to neutrophil membranes, with a concomitant loss from the cytosol. Maximal translocation of all three isoforms occurred by 3 min. No PKC immunoreactivity was observed in a crude nuclear fraction, but PKC-delta and -zeta were found in the granule fraction after degranulation (10 min). PKC activity (Ca2+-dependent and -independent) increased 50- and 19-fold, respectively, by 10 min in the granules from OPZ-stimulated cells. Curiously, no immunoreactive cPKC (alpha and beta(I/II)) could be localized in the granule fraction to account for the Ca2+-dependent PKC activity. Localization of PKC isoforms in the neutrophil membranes and granules suggests their involvement in the regulation of functional responses triggered by OPZ. PKC isoform translocation to membranes from OPZ-stimulated cells preceded both p47phox (a cytosolic component of the NADPH oxidase) translocation and NADPH oxidase assembly. The presence of both PKC isoforms and p47phox in the membrane was transient, with the loss of p47phox occurring sooner than either the loss of membrane-associated PKC or that of NADPH oxidase activity. The apparent EC50 values for PKC translocation and NADPH oxidase assembly were similar. These data suggest that PKC isoforms regulate the assembly and activation of NADPH oxidase induced by OPZ. 相似文献
14.
We have used the fluorescence in situ hybridization (FISH) technique to refine the localization of the cystic fibrosis transmembrane conductance regulator (CFTR) gene on human chromosome 7. The result shows that the gene is most likely located within band q31.3. 相似文献
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Auguste Commeyras Hlne Collet Laurent Boiteau Jacques Taillades Odile Vandenabeele‐Trambouze Herv Cottet Jean‐Philippe Biron Raphaël Plasson Louis Mion Olivier Lagrille Herv Martin Franck Selsis Michel Dobrijevic 《Polymer International》2002,51(7):661-665
Addressing the still open question of the prebiotic origin of sequential macromolecules (peptides, nucleic acids) on the primitive Earth, we describe a molecular engine (the primary pump), which works at ambient temperature and continuously generates, elongates and complexifies sequential peptides. This new scenario is based on a cyclic reaction sequence, whose keystep is the activation of amino acids into their N‐carboxyanhydrides (NCA) through nitrosation by NOx. This process could have taken place on tidal beaches; it requires a buffered ocean, emerged land and a nitrosating atmosphere. With the help of geochemical studies and computer simulations of atmosphere photochemistry, we show that the primitive Earth during the Hadean may have satisfied all these requirements. © 2001 Society of Chemical Industry. 相似文献
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LC Myers MP Terranova AE Ferentz G Wagner GL Verdine 《Canadian Metallurgical Quarterly》1993,261(5125):1164-1167
The Escherichia coli Ada protein repairs methylphosphotriesters in DNA by direct, irreversible methyl transfer to one of its own cysteines. Upon methyl transfer, Ada acquires the ability to bind specific DNA sequences and thereby to induce genes that confer resistance to methylating agents. The amino-terminal domain of Ada, which comprises the methylphosphotriester repair and sequence-specific DNA binding elements, contains a tightly bound zinc ion. Analysis of the zinc binding site by cadmium-113 nuclear magnetic resonance and site-directed mutagenesis revealed that zinc participates in the autocatalytic activation of the active site cysteine and may also function as a conformational switch. 相似文献
19.
LC Kühn 《Canadian Metallurgical Quarterly》1994,7(4):763-785
Cellular iron metabolism comprises pathways of iron-protein synthesis and degradation, iron uptake via transferrin receptor (TfR) or release to the extracellular space, as well as iron deposition into ferritin and remobilization from such stores. Different cell types, depending on their rate of proliferation and/or specific functions, show strong variations in these pathways and have to control their iron metabolism to cope with individual functions. Studies with cultured cells have revealed a specific cytoplasmic protein, called 'iron regulatory protein' (IRP) (previously known as IRE-BP or IRF), that plays a key role in iron homoeostasis by regulating coordinately the synthesis of TfR, ferritin, and erythroid 5-aminolevulinate synthase (eALAS). Present in all tissues analysed, IRP is identical with the [4Fe-4S] cluster containing cytoplasmic aconitase. Under conditions of iron chelation, IRP is an apo-protein which binds with high affinity to specific RNA stem-loop elements (IREs) located 5' of the initiation codon in ferritin and eALAS mRNA, and 3' in the untranslated region of TfR mRNA. At 5' sites IRF blocks mRNA translation, whereas 3' it inhibits TfR mRNA degradation. Both effects compensate for low intracellular iron concentrations. Under high iron conditions, IRP is converted to the holo-protein and dissociates from mRNA. This reverses the control towards less iron uptake and more iron storage. Iron can therefore be considered as a feedback regulator of its own metabolism. It has recently become evident that nitric oxide, produced by macrophages and other cell types in response to interferon-gamma, induces the IRE-binding activity of IRF. Moreover measurements of the RNA-binding activity of IRP in tissue extracts may provide valuable information on iron availability. 相似文献
20.
A new technique for understanding the organization of complex circuits in the vertebrate brain, scanning laser photostimulation, is described. This approach is based on the photolysis of a caged form of the excitatory neurotransmitter glutamate. Computer-controlled photostimulation and whole cell recording in brain slices allow the construction of detailed maps of the position, strength, sign and number of inputs converging on a single postsynaptic neuron. Scanning laser photostimulation offers many advantages over current techniques: spatial resolution is superb, fibers of passage are not activated, and thousands of presynaptic locations can be stimulated. This review describes the technique of photostimulation, outlines the instrumentation, necessary to implement it, and discusses the interpretation of photostimulation-derived data. Several examples of applications, ranging from mapping circuits in the mammalian visual cortex to determining receptor distributions on single neurons are considered. Although still in its early stages, scanning laser photostimulation offers neuroscientists a powerful tool for determining the organization and function of local brain circuits. 相似文献