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991.
Truncation of the human immunodeficiency virus type 1 envelope glycoprotein allows efficient pseudotyping of Moloney murine leukemia virus particles and gene transfer into CD4+ cells. 总被引:1,自引:0,他引:1
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F Mammano F Salvatori S Indraccolo A De Rossi L Chieco-Bianchi H G Gttlinger 《Journal of virology》1997,71(4):3341-3345
Human immunodeficiency virus type 1 (HIV-1) can readily accept envelope (Env) glycoproteins from distantly related retroviruses. However, we previously showed that the HIV-1 Env glycoprotein complex is excluded even from particles formed by the Gag proteins of another lentivirus, visna virus, unless the matrix domain of the visna virus Gag polyprotein is replaced by that of HIV-1. We also showed that the integrity of the HIV-1 matrix domain is critical for the incorporation of wild-type HIV-1 Env protein but not for the incorporation of a truncated form which lacks the 144 C-terminal amino acids of the cytoplasmic domain of the transmembrane glycoprotein. We report here that the C-terminal truncation of the transmembrane glycoprotein also allows the efficient incorporation of HIV-1 Env proteins into viral particles formed by the Gag proteins of the widely divergent Moloney murine leukemia virus (Mo-MLV). Additionally, pseudotyping of a Mo-MLV-based vector with the truncated rather than the full-length HIV-1 Env allowed efficient transduction of human CD4+ cells. These results establish that Mo-MLV-based vectors can be used to target cells susceptible to infection by HIV-1. 相似文献
992.
Elevated AGE-modified ApoB in sera of euglycemic, normolipidemic patients with atherosclerosis: relationship to tissue AGEs. 总被引:9,自引:1,他引:8
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A. W. Stitt C. He S. Friedman L. Scher P. Rossi L. Ong H. Founds Y. M. Li R. Bucala H. Vlassara 《Molecular medicine (Cambridge, Mass.)》1997,3(9):617-627
BACKGROUND: Advanced glycation endproducts (AGEs) are implicated in the pathogenesis of atherosclerotic vascular disease of diabetic and nondiabetic etiology. Recent research suggests that advanced glycation of ApoB contributes to the development of hyperlipidemia. AGE-specific receptors, expressed on vascular endothelium and mononuclear cells, may be involved in both the clearance of, and the inflammatory responses to AGEs. The aim of this study was to examine whether there is a relationship between serum AGE-ApoB and AGEs in arterial tissue of older normolipidemic nondiabetic patients with occlusive atherosclerotic disease, compared with age-matched and younger asymptomatic persons. MATERIALS AND METHODS: Serum AGE-ApoB was measured by ELISA in 21 cardiac bypass patients. Furthermore, an AGE-specific monoclonal antibody, and polyclonal antibodies against anti-AGE-receptor (anti-AGE-R) 1 and 2 were used to explore the localization and distribution of AGEs and AGE-R immunoreactivity (IR) in arterial segments excised from these patients. RESULTS: Serum AGE-ApoB levels were significantly elevated in the asymptomatic, older population, compared with those in young healthy persons (259 +/- 24 versus 180 +/- 21 AGE U/mg of ApoB, p < 0.01). Higher AGE-ApoB levels were observed in those patients with atherosclerosis (329 +/- 23 versus 259 +/- 24 AGE U/mg ApoB, p < 0.05). Comparisons of tissue AGE-collagen with serum AGE-ApoB levels showed a significant correlation (r = 0.707, p < 0.01). In early lesions, AGE-IR occurred mostly extracellularly. In fatty streaks and dense, cellular atheromatous lesions, AGE-IR was visible within lipid-containing smooth muscle cells and macrophages, while in late-stage, acellular plaques, AGE-IR occurred mostly extracellularly. AGE-R1 and -R2 were observed on vascular endothelial and smooth-muscle cells and on infiltrating mononuclear cells in the early-stage lesions, whereas in dense, late-stage plaques, they colocalized mostly with lipid-laden macrophages. On tissue sections, scoring of AGE-immunofluorescence correlated with tissue AGE and plasma AGE-ApoB. CONCLUSIONS: (1) The correlation between arterial tissue AGEs and circulating AGE-ApoB suggests a causal link between AGE modification of lipoproteins and atherosclerosis. AGE-specific receptors may contribute to this process. (2) Serum AGE-ApoB may serve to predict atherosclerosis in asymptomatic patients. 相似文献
993.
Objective
The goal of this study is the evaluation of the functionnal results and the complications of this type of surgery. Such questions have been poorly discussed in the literature.Methods
We report our experience on 80 inflatable prostheses that were implanted between october 1987 and october 1994. The mean follow-up is 3 years and the assessment of the objective (mechanical functioning of the prosthesis and complications) and sujective results (sexuality of the patients) is carried out on 68 patients.Results
The results are the following:994.
Califano Jean-Christophe Goullieux Laurent Amblard Muriel Fehrentz Jean-Alain Bernad Nicole Bergé Gilbert Castel Jean Martinez Jean 《International journal of peptide research and therapeutics》1997,4(4-6):235-239
Summary The involvement of cholecystokinin receptors in the phenomenon of satiety has been the impetus for significant research efforts,
leading to the design and synthesis of CCK-A selective agonists for the possible treatment of obesity. The Abbott laboratories
have described a novel series of pseudotetrapeptides represented by compound A71623, a highly potent and selective peripheral
receptor agonist, but possessing very poor bioavailability. Starting from the structural requirements of this series of compounds,
a peptidomimetic study was investigated, especially focusing on the N-terminal part of A71623. Using standard coupling methods,
introduction of unnatural aromatic amino acids bearing a 2-carboxyethyl side chain on their α-amino group, along with backbone
length modulation, afforded selective analogues, presenting a highly modified peptidic backbone. From our two lead compounds,
further optimization is under development, tending towards nonpeptidic structures. 相似文献
995.
Different thermostabilities of FLP and Cre recombinases: implications for applied site-specific recombination. 总被引:17,自引:3,他引:14
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Genomic manipulations using site-specific recombinases rely on their applied characteristics in living systems. To understand their applied properties so that they can be optimally deployed, we compared the recombinases FLP and Cre in two assays. In both Escherichia coli and in vitro, FLP shows a different temperature optimum than Cre. FLP is more thermolabile, having an optimum near 30 degrees C and little detectable activity above 39 degrees C. Cre is optimally efficient at 37 degrees C and above. Consistent with FLP thermolability, recombination in a mammalian cell line mediated by a ligand- regulated FLP-androgen receptor fusion protein is more efficient at 35 degrees C than at higher temperatures. We also document a mutation in a commercially available FLP plasmid (FLP-F70L) which renders this recombinase even more thermolabile. The different temperature optima of FLP, FLP-F70L and Cre influence their strategies of usage. Our results recommend the use of Cre for applications in mice that require efficient recombination. The thermolabilities of FLP and FLP-F70L can be usefully exploited for gain of function and cell culture applications. 相似文献
996.
The murine lymphomacrophage hybrids ESb, EbF1, EbF2-c4, which express c-fes constitutively, were found by Southern analysis to bear a c-fes deletion of almost 100 bp. The deleted allele was transmitted to the metastatic hybrids by their nonexpressing, poorly metastatic
T-lymphoma progenitor Eb, which also has a structurally normal c-fes allele. PCR amplification and sequencing of fes cDNA spanning exons 3–5, where the deletion mapped, ruled out any involvement of coding sequences in the rearrangement. PCR
amplification of the as yet unsequenced murine c-fes IVS3 and IVS4 showed they are about 50% longer than their human and feline homologs. Sequencing of IVS4 showed no difference
between tumor and control DNA. Sequencing of part of the ∼2600-bp IVS3 was guided by the restriction analysis of PCR products
from control and hybrid DNAs. This showed that differences from the control appeared to be mainly located in the 900-bp HindIII-EcoRI fragment, localized in the middle of IVS3. As all three hybrids had the same restriction map, this fragment was sequenced
in one of them (ESb). A run of >200 CA repeats was found in control DNA, and a reduction in the CAn microsatellite accounted for most of the c-fes deletion in the ESb hybrid. Interestingly, the 50% reduction in the size of human and feline c-fes IVS3 as compared with the murine homolog is mostly due to contraction of the same microsatellite.
Received: 6 November 1995 / Accepted: 2 May 1996 相似文献
997.
The aplocheiloid killifish Rivulus marmoratus is the only known self-fertilizing hermaphroditic vertebrate. Most natural populations consist almost entirely of hermaphrodites and comprise arrays of homozygous clones. However, in almost all populations thus far studied, clonal variation, as detected with molecular techniques, is very high. A karyological survey was carried out on specimens from Brazil, the Bahamas, Belize, and Florida (4 locales) by C-banding, silver staining, and fluorescent staining. The chromosome complement of R. marmoratus is surprisingly uniform over its vast geographic range, in terms of both chromosome number and morphology, heterochromatin distribution and composition, and nucleolar organizer region (NOR) distribution. The short arms of chromosome pair 15, where NORs are located, showed the only variation detected in this study: those of pattern A were consistently shorter than those of pattern B; moreover, the latter show positive heteropycnosis with Giemsa staining. The present data demonstrate that chromosomal variation is not a significant part of the clonal divergence in this species, even though its breeding system, by forming homozygotes for new rearrangements almost immediately, should make that variation easier to detect. The high chromosomal homogeneity is discussed in the light of the peculiar natural history of the species. 相似文献
998.
TheDrosophila larkgene encodes an essential RNA-binding protein of the RNA recognition motif (RRM) class that is required during embryonic development. Genetic analysis demonstrates that it also functions as a molecular element of a circadian clock output pathway, mediating the temporal regulation of adult emergence in the fruitfly. We now report the molecular characterization of a human gene with significant similarity tolark.Based on fluorescencein situhybridization and radiation hybrid mapping, the human gene has been localized to chromosome region 11q13; it is closely linked to several identified genes including the locus of Bardet–Biedl syndrome type 1. Thelark-homologous human gene expresses a single 1.8-kb size class of mRNA in most or all tissues including brain. Additional database searches have identified a mouse counterpart that is virtually identical to the human protein. Similar to lark protein, both mammalian proteins contain two copies of the RRM-type consensus RNA-binding motif. Unlike most RRM family members, however, theDrosophilaand mammalian proteins also contain a retroviral-type (RT) zinc finger that is situated 43 residues C-terminal to the second RRM element. Within a 184-residue segment spanning the RRM elements and the RT zinc finger, the human and mouse proteins are 61% similar to theDrosophilalark sequence. These common sequence features and comparisons among a large collection of RRM proteins suggest that the human and mouse proteins represent homologues ofDrosophilalark. 相似文献
999.
Miroslawa Grzeskowiak Vittorina Della Bianca P. De Togni E. Papini F. Rossi 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1985,844(1):81-90
Exogenous phospholipase C induces in human neutrophils the activation of a respiratory burst, measured as O2 consumption and O2? production and of secretion of specific granules, measured as release of vitamin B-12 binding protein. The secretory response is minimal and follows the onset of the respiratory response. Studies carried out using cells prelabeled with |3H|glycerol and32P on the molecular mechanism of the stimulations demonstrate that the effects are dependent on the formation of diacylglycerol by hydrolysis of different classes of glycerophospholipids. They are, however, independent of the activation of a ‘phosphoinositide turnover’ as occurs in cells stimulated with fMet-Leu-Phe. Furthermore, the respiratory and secretory responses to exogenous phospholipase C are not associated with moditications of cytosolic Ca2+ concentration measured with the Quin-2 method, and the release of bound Ca2+, measured with the membrane probe, chlorotetracycline. Apart from a quantitative difference, mostly regarding the ratio of the intensity of the respiratory and secretory responses, the effects caused by exogenous phospholipase C are qualitative;y similar to those induced by phorbol myristate acetate and are probably linked to an involvement of protein kinase C, activated by diacylglycerol liberated in the plasma membrane. 相似文献
1000.
Ataxia telangiectasia (A-T) cells are sensitive to a broad range of free-radical-producing and alkylating agents. Damage caused by such agents is in part repaired by base excision [base excision repair (BER)]. Two BER pathways have been demonstrated in mammalian cells: a single-nucleotide-insertion pathway and a long-patch pathway involving resynthesis of 2-10 nucleotides. Although early studies failed to detect DNA-repair defects in A-T cells exposed to ionizing radiation and radiomimetic agents, more recent experiments performed in non-dividing A-T cells and the demonstrated interaction of the A-T-mutated protein (ATM) with the BRCA1 gene product suggest that a DNA-repair defect may underlie, at least in part, the radiation sensitivity in A-T cells. We have analysed BER of a single abasic site or a single uracil in two A-T families, using an in vitro BER system. In both families, the mutation involved was homozygous and completely inactivated the ATM protein. No difference was observed between affected individuals and heterozygous or homozygous wild-type relatives in their capacity to perform DNA repair by either one-nucleotide insertion or the long-patch pathway. Hence, the putative DNA-repair defect in A-T cells, if any, does not involve BER. 相似文献