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961.
BACKGROUND: A number of enzymatic techniques have recently been developed to detect DNA fragmentation in apoptosis at the cellular level. However, since DNA fragmentation also occurs in cellular necrosis, we studied to which extent the use of DNA polymerase (nick translation) or terminal transferase (tailing) allows the differentiation between internucleosomal DNA degradation, typical for apoptosis, and the more random DNA destruction in necrosis. EXPERIMENTAL DESIGN: We compared these techniques on in vitro and in vivo models for apoptotic or necrotic cell death. Apoptosis of thymocytes in vitro was induced by gamma-irradiation, necrosis by the cytotoxic action of antibody and complement. Cell death in vivo was examined on paraffin-embedded tissue material from animals with autoimmune encephalomyelitis that served as a model for apoptosis, or in kainic acid-induced nerve cell degeneration as a model for necrosis. RESULTS: DNA fragmentation was visualized by the incorporation of labeled nucleotides into the nuclei of affected cells utilizing tailing or nick translation techniques. In the early stages of cell degeneration in vitro, cells undergoing apoptosis were preferentially labeled by tailing, whereas necrotic cells were identified by nick translation. Similarly, early stages of necrosis in vivo were preferentially detected by nick translation, whereas tailing was slightly more sensitive for the detection of apoptosis. Results obtained with these enzymatic techniques were in accord with the assessment of cell death by morphologic criteria. Both techniques could be applied in tissue samples even after prolonged fixation in paraformaldehyde if the sections were pretreated with proteinase K digestion. CONCLUSIONS: Our studies show that both in situ nick translation and in situ tailing are useful in detecting DNA fragmentation in cell suspensions and tissue sections. These techniques may help to define the molecular mechanisms leading to cell death in experimental conditions and eventually in human tissue. 相似文献
962.
The cytokine macrophage migration inhibitory factor (MIF) is a major protein constituent of the anterior pituitary gland released into the bloodstream during endotoxaemia. For many years, MIF had been thought to be a T cell product associated with delayed-type hypersensitivity reactions. The identification of MIF as a pituitary 'stress' hormone provides an important link in the regulation of systemic inflammatory responses by the central nervous system. 相似文献
963.
964.
PM Warnecke D Biniszkiewicz R Jaenisch M Frommer SJ Clark 《Canadian Metallurgical Quarterly》1998,22(2):111-121
Currently, the pig species is regarded as the most likely organ donor for human xenotransplantation in the future. However, it cannot be granted that the pig will be the optimal species of choice. We have studied human anti-sheep antibodies in comparison with anti-pig antibodies. The anti-sheep lymphocytotoxic and hemagglutination titers were in the range 8 to 128 and 2 to 32, respectively, in single individuals, which were considerably lower than the anti-pig titers of these individuals. Perfusion of sheep kidneys with human blood reduced the anti-sheep xenoantibody titers to zero as measured by lymphocytotoxic, hemagglutination, and sheep aortic endothelial cell antibody binding assays. The perfused kidneys showed generalised depositions of human IgM and C3c in the vascular tree and focal depositions of C1q and fibrin. Obliteration of capillaries by human platelets and polymorphonuclear cells were observed. Total neutral glycolipid fractions were isolated from sheep intestinal, pancreatic, and kidney tissues. By using a chromatogram binding assay, a monoclonal anti-Forssman antibody identified a single compound with five sugar residues in all organs. Several glycolipid bands were stained in all organs by the Gal(alpha)1-specific lectin I-B4 from Griffonia (Bandeiraea) Simplicifolia. A human AB serum pool showed staining by both IgG and IgM antibodies of the Forssman and Gal(alpha)1-terminating components as well as some other, not structurally identified, components. The Forssman and Gal(alpha)1-reactivity in human sera could be eliminated by immunoadsorption using Forssman and Gal(alpha)1-3Gal-immunoadsorbent columns, respectively. Immunostaining of sheep kidney tissue sections showed the presence of Gal(alpha)1-terminating epitopes by immunoperoxidase and immunogold silver staining techniques. Proximal convoluted tubules showed a strong staining, while thin loops of Henle, collecting ducts, urothelium, and vessels showed a weaker staining. Distal convoluted tubules and thick loops of Henle were completely negative. In summary, human serum contains anti-sheep xenoantibodies reacting mainly with the Forssman and Gal(alpha)1-determinants in sheep tissues and the anti-sheep antibody titers are lower than the corresponding anti-pig titers. 相似文献
965.
966.
S Gilfillan M Bachmann S Trembleau L Adorini U Kalinke R Zinkernagel C Benoist D Mathis 《Canadian Metallurgical Quarterly》1995,25(11):3115-3122
Mice with a null mutation in the terminal deoxynucleotidyl transferase (TdT) gene harbor immunoglobulin and T cell receptor repertoires essentially devoid of N-region diversity. Consequently, the CDR3 loops important for antigen recognition are shorter and considerably less diverse than those of wild-type controls. We find surprisingly normal immune responses in TdT0 mice, as regards both efficiency and specificity. This provokes a reconsideration of the assumption that N-region diversity is required for an effective T and B cell repertoire. 相似文献
967.
The powder mixtures used for diffusion galvanizing (Zn + inert filler + activator) have the following deficiencies: susceptibility to sintering and fusing under elevated CTT temperatures, complexity of correcting the set of components in repeated use of the mixtures and the related instability of the results obtained, complexity of regeneration of worked-out mixtures, elevated ecological hazard of their use, etc. This explains the need for the present work, which shows that most of the mentioned deficiencies can be eliminated by an appropriate choice of the mixture components and (or) preliminary treatment of them. 相似文献
968.
R Spanagel 《Canadian Metallurgical Quarterly》1995,70(1):37-49
Behavioural sensitization involves progressive increases in behavioural responses to repeated intermittent administration of drugs of abuse. Behavioural sensitization is observed to the locomotor stimulant, rewarding and discriminative effects of a drug. These are effects which seem to be essential in the initiation, expression and maintenance of a drug-seeking behaviour. Therefore the phenomenon of behavioural sensitization may have important implications for the understanding of addictive processes. Findings given in this review demonstrate the involvement of endogenous opioid systems in the initiation of sensitized responses on the neurochemical level, i.e., within the mesolimbic dopaminergic system, as well as on the behavioural level. Specifically, it is shown that behavioural sensitization to morphine and cocaine is modulated by endogenous kappa-opioid systems. 相似文献
969.
970.
It is argued that the technology needed to support the emerging broadband integrated services digital network technology (B-ISDN) is on the horizon, but the deployment of this technology in the telecommunication plant will not be sufficient to make B-ISDN a commercial success. Two corequisites for ensuring the long-term success of the emerging B-ISDN include stimulating the demand for bandwidth by inventing and promoting new telecommunication applications, and enabling the interconnection or convergence of the emerging high-speed network and the existing cable television infrastructure. The author believes that this is necessary because no alternative seems to offer a workable base for financing the transition to B-ISDN and for reaching the needed economy of scale, and because the intrinsic value of the public network grows combinatorially with the number of subscribers 相似文献