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51.
Stephanie Gordon Phillips Shiu-Lan Lui David M. Phillips 《In vitro cellular & developmental biology. Plant》1982,18(8):727-738
Summary Epithelial cells may relate to their basement membrane substrates via lectin-like interactions. In a model system for study
of this type of interaction, lectin-coated bacteriological plastic petri dishes were presented as substrates for epithelial
cell adhesion. Of 21 lectins tested by mixed agglutination against two epithelial cell types, Madin-Darby canine kidney (MDCK),
and human embryonic kidney cells (HEK), nine gave less than 5% rosettes and 12 gave 5 to 50% rosettes. Wheat germ agglutinin
(WGA) andGeodia cydonium lectin gave the highest percentage of rosettes. Wheat germ agglutinin was readily adsorbed to plastic surfaces and maintained
specificity in binding interactions. Both MDCK and HEK cells attached as well to WGA coated petri dishes as to conventional
tissue culture dishes. Furthermore, both spread over the lectin-coated surfaces. The MDCK cells grew to confluence and could
be subcultured and maintained indefinitely on such surfaces, although WGA in solution was toxic to the cells in concentrations
as low as 0.1 to 1.0 μg/ml. Cell attachment to WGA coated dishes was blocked by cycloheximide only if the cells had been preincubated
with the inhibitor for several hours. Cell attachment was not inhibited by pretreatment of cells with neuraminidase. Precoating
cells with WGA blocked binding to both WGA-coated surfaces and untreated tissue culture dishes. Cells attached to WGA-coated
dishes could not be readily dislodged by trypsin-EDTA for the first 2 h after subculture. By 4 h, attachment was again trypsin
sensitive, suggesting that the cells synthesized a trypsin-sensitive material that was laid down between the cell surface
and the WGA-coated dish. Regeneration of trypsin sensitivity was not blocked by cycloheximide.
This work was supported by Research Grant AG01986 from the National Institutes of Health, Bethesda, Maryland. 相似文献
52.
Synthetic leukotrienes (LT) C4 and D4 elicited concentration-dependent contractions of the guinea pig uterus between 10?8-10?6M, whereas LTE4 appeared 1000-fold weaker. The potencies of LTC4 and LTD4 were similar to that of acetylcholine and PGF2α but weaker than that of PGE2. The maximal contractions elicited by LTC4 and LTD4 were 66.0 ± 2.1% and 63.8 ± 4.6% that elicited by acetylcholine. FPL 55712 (10?5M) antagonized the uterine contractile activity of LTD4, while meclofenamic acid at 10?5M but not at 10?6M also antagonized the LTD4-induced contration. Radioimmunoassay of the uterine tissue bathing fluid following LTD4 indicated the variable presence of low concentrations of PGE2, PGF2α and TXB2. These results demonstrate the LTC4 and LTD4 possess significant uterine contractile activity, which may only partially be mediated indirectly via prostaglandin products. 相似文献
53.
Philip L. Smith Stephanie A. Orellana Michael Field M.D. 《The Journal of membrane biology》1981,63(3):199-206
Summary Unidirectional fluxes of35SO4 across and into rabbit ileal epithelium were measured under short-circuit conditions, mostly at a medium SO4 concentration of 2.4mm. Unidirectional mucosa (m)-to-serosa (s) ands-to-m fluxes (J
ms,J
sm) were 0.456 and 0.067 moles hr–1 cm–2, respectively.J
ms was 2.7 times higher in distal ileum than in mid-jejunum. Ouabain abolished net SO4 transport (J
net) by reducingJ
ms. Epinephrine, a stimulus of Cl absorption, had no effect on SO4 fluxes. Theophylline, a stimulus of Cl secretion, reducedJ
ms without affectingJ
sm, causing a 33% reduction inJ
net. Other secretory stimuli (8-Br-cAMP, heat-stable enterotoxin, Ca-ionophore A23187) had similar effects. Replacement of all Cl with gluconate markedly reducedJ
net through both a decrease inJ
ms and an increase inJ
sm. The anion-exchange inhibitor, 4-acetoamido-4-isothiocyano-2,2-sulfonic acid stilbene (SITS), when added to the serosal side, reducedJ
ms by 94%, nearly abolishingJ
net. SITS also decreasedJ
sm by 75%. Mucosal SITS (50 m) was ineffective. 4,4-diisothiocyano-2,2-sulfonic acid stilbene (DIDS) had effects similar to SITS but was less potent. Measurements of initial rates of epithelial uptake from the luminal side (J
me) revealed the following: (1)J
me is a saturable function of medium concentration with aV
max of 0.94 moles hr–1 cm–2 and aK
1/2 of 1.3mm; (2) replacing all Na with choline abolishedJ
me; (3) replacing all Cl with gluconate increasedJ
me by 40%; (4) serosal SITS had no effect onJ
me; and (5) stimuli of Cl secretion had no effect onJ
me or increased it slightly. Determination of cell SO4 with35SO4 indicated that, at steady-state, the average mucosal concentration is 1.1 mmoles per liter cell water, less than half the medium concentration. Cell SO4 was increased to 3.0mm by adding SITS to the serosal side. Despite net transport rates greater than 1.4 Eq hr–1 cm–2, neither addition of SO4 to the SO4-free medium nor addition of SITS to SO4-containing medium altered short-circuit current. The results suggest that (1) ileal SO4 absorption consists of Na-coupled influx (symport) across the brush border and Cl-coupled efflux (antiport) across the basolateral membrane; (2) the overall process is electrically neutral; (3) the medium-to-cell Cl concentration difference may provide part of the driving force for net SO4 absorption; and (4) since agents affecting Cl fluxes (both absorptive and secretory) have little effect on SO4 fluxes, the mechanisms for their transcellular transports are under separate regulation. 相似文献
54.
55.
Robert L. Wykle Craig H. Miller Jon C. Lewis Jeffrey D. Schmitt Jennie A. Smith Jefferson R. Surles Claude Piantadosi Joseph T. OFlaherty 《Biochemical and biophysical research communications》1981,100(4):1651-1658
1-O-Hexadecyl-2-O-acetyl--glycero-3-phosphocholine (platelet activating factor) stimulated the degranulation of rabbit platelets and human neutrophils, whereas the enantiomer, 3-O-hexadecyl-2-O-acetyl--glycero-1-phosphocholine, was inactive. The analogs compared had the following relative potencies in degranulating platelets and neutrophils: 1-O-hexadecyl-2-O-acetyl--glycero-3-phosphocholine > 1-O-hexadecyl-2-O-ethyl--glycero-3-phosphocholine >-1-O-octadecyl-2-O-ethylglycero-3-phosphocholine = 1-O-hexadecyl-2-O-methyl--glycero-3-phosphocholine >-1-O-dodecyl-2-O-ethyl-glycero-3-phosphocholine. The deacetylated compound, 1-O-hexadecyl-2-lyso--glycero-3-phosphocholine, and 1-O-hexadecyl-2,2-dimethylpropanediol-3-phosphocholine were inactive. The active analogs selectively desensitized the response to each other in the neutrophils. It is suggested that these compounds may activate cells through interaction with a stereospecific receptor. 相似文献
56.
Jacqueline S. Lipszyc Stephanie Gordon Phillips Orlando J. Miller 《Experimental cell research》1981,133(2)
Autoradiographic analysis of [3H]uridine incorporation 48 h after polyethylene glycol-mediated cell fusion indicates that nucleolar RNA synthesis persists in both human and mouse nuclei in interspecific heterokaryons. The absence of nucleolar dominance in heterokaryons has been confirmed by zinc-dithizone nucleolus-specific staining, and is true even when there are considerably more nuclei of one species than of the other in the heterokaryon. Studies of actinomycin D-induced nucleolar segregation indicate that the zinc-binding proteins responsible for zinc-dithizone staining are located in a different nucleolar component than the protein responsible for silver staining. 相似文献
57.
Effect of wind-borne pollens on the population dynamics ofAmblyseius hibisci [Acarina: Phytoseiidae]
Populations ofAmblyseius hibisci (Chant), an important predator of citrus red mite,Panonychus citri (McGregor) exhibited varying responses to field applications of cat-tail,Typha latifolia L., pollen in San Joaquin Valley citrus groves. Predator densities increased to significantly higher levels after pollen treatments during the fall months, and to a limited extent after late winter treatments, but not after early spring treatments. The amount of response appeared to be inversely related to the levels of natural foods present during the various seasons. Results of the wind-borne pollen monitoring studies indicated that population growth inA. hibisci was correlated with the seasonal pattern of pollen fallout. Population growth was slowest in years when pollen catches were low during the late winter and early spring, and fastest in years when peak pollen catches occurred in late winter or early spring. Population growth inA. hibisci did not appear to be markedly responsive toP. citri populations although suppression of prey densities was indicated in ca. 50% of the cases. In laboratory tests,A. hibisci reproduced through 3 generations on 11 of 23 pollens tested. 相似文献
58.
Stephanie G. Phillips David M. Phillips V. G. Dev Dorothy A. Miller O. P. Van Diggelen O. J. Miller 《Experimental cell research》1976,98(2)
Electron microscopic evidence suggests that sperm can be spontaneously incorporated by cultured cells but cytogenetic and biochemical evidence indicate that sperm do not introduce new genes into such cells with detectable frequency. Sperm suspensions from mouse or Chinese hamster epididymis or human semen were added to cultures of RAG, a mouse cell line which dies in HAT medium because of HPRT deficiency. In EMs, sperm appeared to be readily phagocytized and degraded by the cells. When sperm-treated cultures were transferred to HAT medium resistant clones arose at a frequency of about 10−6, or at least 25× the reversion rate of RAG. Most HAT-resistant clones had HPRT activity which migrated electrophoretically like HPRT of the sperm donor species, though one was apparently a spontaneous RAG revertant. Most HAT-resistant clones had some chromosomes of the sperm donor species. In human sperm× RAG clones, the array of human chromosomes suggested that the human parent had been diploid rather than haploid; some cells contained both homologues of a polymorphic pair and some contained both X and Y. Furthermore, some sperm suspensions plated alone into flasks generated colonies, thus revealing the presence of low numbers of viable somatic cells. Presence of contaminating somatic cells in a sperm suspension was correlated with ability to induce HAT-resistant colonies when the suspension was added to RAG cells. Taken together, the data suggest that correction of the HPRT deficiency of RAG by sperm suspensions occurs at very low frequency and is probably due to efficient spontaneous fusion of low numbers of contaminating somatic cells with RAG cells. 相似文献
59.
Skin and tumor grafting experiments performed on F1 hybrid mice indicate that theH(Tla) histocompatibility system is composed of at least two genetic loci,H(Tla-1) andH(Tla-2), and that one of these loci,H(Tla-1), has at least three alleles. We suggest thatH(Tla-1) andH(Tla-2) be renamedH-31 andH-32, respectively. 相似文献
60.
Scott Davis Stephanie Propp Susan M. Freier Laura E. Jones Martin J. Serra Garth Kinberger Balkrishen Bhat Eric E. Swayze C. Frank Bennett Christine Esau 《Nucleic acids research》2009,37(1):70-77
Chemically modified antisense oligonucleotides (ASOs) are widely used as a tool to functionalize microRNAs (miRNAs). Reduction of miRNA level after ASO inhibition is commonly reported to show efficacy. Whether this is the most relevant endpoint for measuring miRNA inhibition has not been adequately addressed in the field although it has important implications for evaluating miRNA targeting studies. Using a novel approach to quantitate miRNA levels in the presence of excess ASO, we have discovered that the outcome of miRNA inhibition can vary depending on the chemical modification of the ASO. Although some miRNA inhibitors cause a decrease in mature miRNA levels, we have identified a novel 2′-fluoro/2′-methoxyethyl modified ASO motif with dramatically improved in vivo potency which does not. These studies show there are multiple mechanisms of miRNA inhibition by ASOs and that evaluation of secondary endpoints is crucial for interpreting miRNA inhibition studies. 相似文献