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11.
Purification and properties of a human plasma endogenous modulator for the platelet tricyclic binding/serotonin transport complex 总被引:1,自引:0,他引:1
An endogenous modulator for the site labeled by [3H]imipramine which is putatively coupled to the serotonin transporter in human platelets was isolated and purified from plasma. Procedures included sequential chromatography on Cibacron blue-Sepharose 4B, concanavalin A-Sepharose 4B, Mono Q HR 10/10 anion exchange, DuPont GF-250 gel permeation and Mono S HR 5/5 cation exchange columns. The purified modulator is a protein of Mr 45,000 with a very acidic pK (less than 3) and sensitive to various proteinases but heat- and acid-stable. This protein inhibited [3H]imipramine binding to platelet membranes competitively (IC50 approximately 6 microM) and enhanced serotonin uptake in fresh human platelets (EC50 approximately 7 microM). Various physicochemical properties, including chromatographic, electrophoretic and immunological as well as amino acid composition analysis revealed that the isolated protein is most probably the human alpha 1-acid glycoprotein. 相似文献
12.
Hemorrhage in mice produces alterations in B cell repertoires 总被引:1,自引:0,他引:1
Multiple organ system failure secondary to infection is the major cause of late deaths after trauma and hemorrhage. The production by B cells of antibodies directed against bacterial antigens is an important component of host defenses. In order to determine the effects of hemorrhage on B cell function, we examined hemorrhage-induced alterations in available (clonal precursors) and actual (plasma cells) B cell repertoires in the course of an immune response toward bacterial antigens. Hemorrhage produced greater than twofold decreases in the absolute frequency and number of clonal precursors specific for the bacterial antigens dextran, levan, and pneumococcal polysaccharide type II. After blood loss, there were decreases in absolute frequency, but not in numbers, of clonal precursors capable of producing antibodies against the nonbacterial antigens ovalbumin and mouse transferrin. Immunization with the bacterial antigen levan within 24 hr of hemorrhage resulted in approximately 50% fewer levan-specific plasma cells than that seen in normal, unhemorrhaged mice. These results demonstrate that hemorrhage produces marked alterations in B cell repertoires, which may contribute to postinjury abnormalities in host defenses. 相似文献
13.
A circadian feeding rhythm which may be entrained by photoperiod was found in fifth instar caterpillars of the lepidopteran, Achaea janata (L.). Allatectomy reduced the amount of food consumed; this consumption was significantly lower during the fifth instar in both allatectomized and sham-operated insects. An apparent circadian feeding pattern appeared on day 2 in the sham-operated caterpillars. Topical application of the anti-allatin, Precocene-II (50 micrograms/animal) also reduced leaf consumption significantly compared to the respective controls, although these controls maintained the same apparent circadian feeding rhythm on day 2. 相似文献
14.
Immunochemical identification of the serine protease inhibitor alpha 1-antichymotrypsin in the brain amyloid deposits of Alzheimer's disease 总被引:42,自引:0,他引:42
Two approaches--molecular cloning and immunochemical analysis--have identified one of the components of Alzheimer's disease amyloid deposits as the serine protease inhibitor alpha 1-antichymotrypsin. An antiserum against isolated Alzheimer amyloid deposits detected immunoreactivity in normal liver. The antiserum was then used to screen a liver cDNA expression library, yielding three related clones. DNA sequence analysis showed that these clones code for alpha 1-antichymotrypsin. Antisera against purified alpha 1-antichymotrypsin stained Alzheimer amyloid deposits, both in situ and after detergent extraction from brain. The anti-amyloid antiserum recognizes at least two distinct epitopes in alpha 1-antichymotrypsin, further supporting the presence of this protein in Alzheimer amyloid deposits. In addition to being produced in the liver and released into the serum, alpha 1-antichymotrypsin is expressed in Alzheimer brain, particularly in areas that develop amyloid lesions. Models by which alpha 1-antichymotrypsin could contribute to the development of Alzheimer amyloid deposits are discussed. 相似文献
15.
Carp were maintained for a month in well-oxygenated water fertilized with organic manure. During the experiment the thickness of the epidermis increased from 140 to 180 urn. Fish from pulluted water were dark, an adaptation to the dark, turbid water. The number of cytoplasmic extensions from the dermal pigment cells increased continuously from 6 per unit length in control specimens to over 80 in the experimental specimens at the end of the month. Apart from background adaptation, this activity of the pigment cells may be a stress reaction. Holocrine secretion of mucous cells was pronounced, with a progressive reduction on the first day after the transfer, to almost total disappearance of this cell type from the epidermis after 3 days. A thick mucous coat became visible on the outside of the epidermis. Eight days after the transfer, a slightly subnormal mucous cell count was observed, indicating the development of newly differentiated mucous cells. This subnormal cell count lasted until the end of the experiment. The pavement cells actively secreted glycocalyx, while newly differentiated pavement cells with still-intact secretory granules replaced the exhausted cells at the epidermis surface throughout the experimental period. Granulocytes, both baso- and neutrophilic, as well as macrophages, infiltrated the epidermis; despite the high bacterial count in the water, no bacteria were observed either inside the skin or entangled in the mucous coat. 相似文献
16.
A novel collagen-glycosaminoglycan (C-GAG) substrate was developed to overcome the optical opacity of a HATF nitrocellulose substrate and to provide a more physiological permeable substrate for cocultured Sertoli and spermatogenic cells. Cocultures were prepared on optically transparent C-GAG discs attached to a polyester mesh to facilitate handling. Sertoli cells displayed a cuboidal-to-columnar shape; a large number of spermatogonia and primary spermatocytes connected by intercellular bridges were associated with basolateral and apical surfaces of Sertoli cells up to 12 days after plating. Rat Sertoli-spermatogenic cell cocultures have been used for testing the effect of toxicants on rat spermatogenesis in vitro. In our initial studies, we tested the effects of the toxicant gossypol on spermatogenic cells cocultured with Sertoli cells on nonpermeable (plastic) and permeable substrates (HATF nitrocellulose) under both standard culture conditions and during perifusion after achieving a continuous electrical-resistant cell monolayer. A selective mitochondrial structural damage was observed in spermatogenic cells (spermatogonia and spermatocytes) but not in the coexisting Sertoli cells. This damage was time- (15–60 min) and dose-dependent (0.1–10µM) and developed more rapidly under perifusion conditions. Similar mitochondrial damage was reported in the intact animal but required higher concentrations (mg) and longer administration time (months) for detection. Studies are in progress to evaluate the effect of additional toxic chemical agents on functional properties of Sertoli and spermatogenic cells in cocultures prepared on various classes of C-GAG substrates.Abbreviations C-GAG,
collagen type I-glycosaminoglycans
- C-C6S,
collagen type I-chondroitin-6-sulfate
- C-H,
collagen type I-heparin 相似文献
17.
Regulation of T cell receptor signaling by a src family protein-tyrosine kinase (p59fyn) 总被引:77,自引:0,他引:77
Engagement of the clonotypic antigen receptor (TCR) on T lymphocytes provokes an activation response leading to cell proliferation and lymphokine secretion. To examine the molecular basis of T cell signaling, we generated transgenic animals in which a lymphocyte-specific nonreceptor protein-tyrosine kinase p59fyn(T) is 20-fold overexpressed in developing T lineage cells. Thymocytes from these mice, analyzed using both cellular and biochemical assays, were remarkably hyperstimulable. Moreover, the responsiveness of normal thymocytes to TCR-derived signals correlated well with the extent to which p59fyn was expressed in these cells. Overexpression of a catalytically inactive form of p59fyn substantially inhibited TCR-mediated activation in otherwise normal thymocytes. These effects are unique to p59fyn; overexpression of a closely related T cell-specific tyrosine kinase, p56lck, elicits dramatically different phenotypes. Our results suggest that p59fyn is a critically important component of the TCR signal transduction apparatus. 相似文献
18.
W.M. Abraham S. Lanes A. Wanner J.S. Stevenson E. Codias L.D. Yerger 《Prostaglandins & other lipid mediators》1986,31(3)
Allergic sheep respond to inhaled
antigen with either acute and late bronchial obstructions (dual responders) or only acute bronchoconstriction (acute responders). In this study we tested the hypothesis that one factor which may distinguish between these two populations is the difference in sensitivity to a specific mediator of airway anaphylaxis, leukotriene (LT) D4 (a major component of slow reacting substance of anaphylaxis). We postulated that if the hypothesis was correct than dual responders should demonstrate increased airway responses to inhaled LTD4 and that this increased responsiveness should also be reflected by a more severe response to inhaled antigen. To test this we used animals from both groups with the same degree of non-specific airway responsiveness to carbachol and determined their airway responses to controlled inhlation challenges with synthetic LTD4 and
antigen. Airway responsiveness to carbachol was determined by measuring the change in specific lung resistance (SRL) to increasing concentrations of carbachol aerosol, and then identifying, by linear interpolation, the provocative carbachol concentration which produced a 150% increase (PC150) in SRL. Airway responses to LTD4, and antigen were determined by measuring the percentage change in SRL after a controlled inhlation challenge with either aerosol. Airway responsiveness to carbachol was not different between the two groups. There was, however, a difference (p<0.05) in the airway response to the same dose of LTD4 in the two groups. Dual responders showed a 297±72% increase in SRL as compared to a 90±13% increase in SRL in the acute responders. Dual responders also showed a greater immediate and more prolonged response to antigen than did acute responders. These results suggest that increased responsiveness to LTD4 may be one factor which may distinguish dual responders from acute responders. 相似文献
19.
DNA-mediated gene transfer of a mutant regulatory subunit of cAMP-dependent protein kinase 总被引:3,自引:0,他引:3
I Abraham S Brill J Hyde R Fleischmann M Chapman M M Gottesman 《The Journal of biological chemistry》1985,260(26):13934-13940
We have used DNA-mediated gene transfer of genomic DNA to introduce into wild-type Chinese hamster ovary (CHO) cells a mutant gene that confers resistance to the growth inhibitory effect of cAMP. This dominant mutation in CHO cell line 10248 is responsible for an alteration in the RI subunit (RI*) of the type I cAMP-dependent protein kinase (Singh, T. J., Hochman, J., Verna, R., Chapman, M., Abraham, I., Pastan, I.H., and Gottesman, M.M. (1985) J. Biol. Chem. 260, 13927-13933). The transformant 11564 which was studied in detail, has the same characteristics as the original mutant 10248 including continued growth in medium containing 8-Br-cAMP, an increase in the Ka for cAMP activation of the kinase, a greatly reduced amount of type II protein kinase activity, an altered incorporation of the photoaffinity label 8-N3[32P]cAMP into the RI* subunit of PKI, and an absence of cAMP-dependent phosphorylation of a Mr = 52,000 protein in intact cells. In addition, analysis of the DNA of the transformant indicates the presence of an increased amount of DNA for the RI gene. These results are consistent with the transfer of a mutant gene for the RI* subunit of the cAMP-dependent protein kinase and its phenotypic expression in the transformant and also support the hypothesis that the mutation responsible for the defect in cell line 10248 is due to an alteration in the gene for RI. 相似文献
20.
Jörg U. Ganzhorn Jean Prospère Abraham Marlène Razanahoera-Rakotomalala 《Primates; journal of primatology》1985,26(4):452-463
Two groups ofAvahi laniger were studied in the Forêt de Analamozoatra near Perinet in the eastern rainforest of Madagascar from August to October 1984.
Overlap between the home ranges of neighbouring groups ofA. laniger was minimal. Group size ranged from one to four individuals with a median group size of two. In four out of ten groups a
baby was born between August and September.A. laniger were most active after dusk and before dawn. They had an extended resting period around midnight. Their diet consisted mostly
of leaves from at least 17 different plant species. They also ate flowers. Fruit eating was recorded twice. Leaves eaten had
high contents of protein and sugar but did not contain alkaloids. The concentration of condensed tannins did not differ between
food items and non-food items. There was no indication of competition with other prosimians that might explain their nocturnality. 相似文献