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61.
Leonard F. Liebes Howard Fleit Dorothea Zucker-Franklin Robert Silber 《Biochimica et Biophysica Acta (BBA)/General Subjects》1980,633(2):245-257
Tubulin has been purified from human blood and tonsil lymphocytes. Using gel filtration, the molecular weight of human lymphocyte tubulin was estimated to be 119 000. The proteins was shown to consist of two subunits, with molecular weights of 61 000 and 58 000 comparable to the α and β polypeptides of human brain tubulin. A partial identity reaction was observed between lymphocyte tubulin and human tubulin when tested by double immunodiffusion against a rabbit anti-human brain tubulin antibody. In the presence of GTP, the purified protein polymerized to form microtubules. Tubulin was localized to the cell's juxtacentriolar region by immunofluorescence and electron microscopy. When assayed by a colchicine-binding assay corrected for time decay, the binding affinity was 1.50 ± 0.86 · 106M?1 and a level in normal lymphocytes of 1.21 · 10?2 ± 0.79 g/g of soluble protein was determined. Since chronic lymphocytic leukemia lymphocytes have an anomalous capping behavior as well as an unusual susceptibility to colchicine toxicity, the properties and levels of tubulin were determined in these cells. Similar values were obtained for the level, decay rate, molecular weight, and Ka for colchicine as for normal lymphocytes. Chronic lymphocytic leukemia lymphocyte tubulin polymerized in a normal fashion. It thus appears that a decrease in the quantity or function of tubulin does not account for these anomalies in the chronic lymphocytic leukemia lymphocyte. 相似文献
62.
Barbara D. Abbott Robert M. Pratt 《In vitro cellular & developmental biology. Plant》1988,24(4):343-352
Summary Retinoids and growth factors seem to be important for normal mammalian reproduction and development. High levels of retinoic
acid are teratogenic and induce cleft palate in the mouse. Little is known concerning the mechanisms through which retinoids
induce cleft palate. Palatal epithelia from CD-1 embryonic mice on Day 12 of gestation were isolated from the mesenchyme and
cultured in serum-free media, with all-trans retinoic acid or 13-cis retinoic acid, with or without epidermal growth factor
(EGF). The epithelia attached and grew, and the cells differentiated over a 72-h culture period. Binding of [125I]EGF was observed in all cultures in a pattern that correlated with thymidine (TdR) uptake by the epithelia. EGF enhanced
growth and [3H]TdR incorporation of the oral cells, but nasal cells generally did not proliferate. In this culture system, both retinoids
suppressed [3H]TdR incorporation in a concentration-dependent manner for epithelia cultured with or without EGF. Medial cells are important
to normal palatogenesis as they play a role in fusion of opposing shelves and subsequently many of these cells undergo programmed
cell death. Death of medial cells in vitro is prevented by EGF and by the retinoids, either with or without EGF. This response
occurs in the absence of a mesenchymal interaction, suggesting that the medial cell response to EGF and retinoids is not mediated
by or dependent on the mesenchymal tissues. The survival of medial cells may be responsible for the failure of opposing shelves
to fuse. 相似文献
63.
64.
Summary Pollen mother cells at early meiotic prophase fromFritillaria lanceolata, F. mutica, Tulbaghia violacea, the lily “Formobel”,Triticum aegilopoides, T. dicoccoides, T. aestivum and synaptic and asynaptic forms ofT. durum were studied in thin sections with the electron microscope (a) in relation to distribution of nuclear pores (b) in respect
of fine structure of the pore complex in those of the first four. The pores were distributed in random clusters during leptotene
to pachytene in all plants, except in the two forms ofT. durum where there were either no pores or so few that they were not detectable. Probably correlated with this, the two membranes
of the nuclear envelope were often widely separated and frequently sacculated. No pores were seen at leptotene in the part
of the envelope to which, in theFritillarias and lily, the nucleolus was adpressed at this time. Evidence supporting a recent model which proposes that annuli are composed
of three rings of eight granular subunits was obtained. These subunits as well as a dense central element, observed in most
pores, were composed of filaments about 3 nm in diameter and evidently protein in character. There was evidence of a continuity
between filaments in the central element and those in the rings of subunits which encircle the pore aperture at both the nuclear
and cytoplasmic sides of the pore. In profiles of pores knobbed filaments were sometimes seen extending laterally from the
pore wall into the perinuclear space at two sides. Questions concerning the role of the annulus are discussed.
The author wish to thank Mr. R. F. Scott for construction to the model. 相似文献
65.
66.
The imaginal pore plates of Hymenoptera apocrita so far examined embody five or six envelope cells respectively. In early developmental stages, however, supernumerary envelope cells have been found. The results are discussed in the context of cell death as a developmental phenomenon. 相似文献
67.
68.
The effects of two glycosylated whey hydrolysates (GWH-Gal A and GWH-Gal B) on glutathione (GSH) and related antioxidant enzymes in SGC-7901 cells were evaluated. Two whey glycosylated hydrolysates promoted an increase in reduced glutathione (GSH) in normal SGC-7901 cells. GSH, glutathione peroxidase (GPx), γ-glutamine cysteine synthetaase (γ-GCS), and catalase (CAT) at 1.0 and 2.0 mg/mL in normal SGC-7901 cells were higher in the GWH-Gal A group than in the GWH-Gal B group (P < 0.05). Compared with GWH-Gal B, GWH-Gal A more strongly inhibited decreases in intracellular GSH, GPx, γ-GCS, CAT, and superoxide dismutase (SOD) in H2O2-induced SGC-7901 cells. Compared with GWH-Gal B, GWH-Gal A at 1.0 and 2.0 mg/mL effectively inhibited increases in lactate dehydrogenase (LDH) and malondialdehyde (MDA) in H2O2-induced SGC-7901 cells (P < 0.05). Therefore, GSH content and related antioxidant enzyme activity levels (GPx, γ-GCS, CAT, SOD) in both normal and H2O2-induced SGC-7901 cells were considerably stronger in the GWH-Gal A group than in the GWH-Gal B group. 相似文献
69.
Summary AHG-associated protein (AHG-a.p.), the antigen of the blood-clotting factor VIII complex, is a specific endothelial cell marker. Primary (p-XTH) and established (XTH-2) endothelial cells from the hearts of Xenopus laevis tadpoles were assayed for the presence of this marker by means of immunological cross-reaction (recognition of common antigenic sites) with antiserum against human AHG-a.p. Radial imtnunodiffusion and rocket immunoelectrophoresis proved to be insufficiently sensitive, whereas immunofluorescence and a newly evaluated ELISA technique gave positive results. The very high sensitivity of the ELISA (less than 1/240000 of the AHG-a.p. in 0.1 ml human standard plasma can be detected) and the removal of interfering proteins by gel filtration also revealed the presence of AHG-a.p. in the fetal calf serum used in the culture medium; earlier investigations into this subject by a one-step radioimmunoassay had reported negative results. Specially adapted XTH-2 cells were grown in a proteinand serum-free hydrolysate medium in order to demonstrate the presence of a Xenopus-derived antigen that was immunoreactive with the anti-human AHG-a.p. 相似文献
70.
Jin-Young Park 《Analytical biochemistry》2009,394(2):217-6489
Transglutaminases (TGs), a family of calcium-dependent transamidating enzymes, are involved in functions such as apoptosis and inflammation and play a role in autoimmune diseases and neurodegenerative disorders. In this study, we describe a novel array-based approach to rapidly determine in situ TG activity in human umbilical vein endothelial cells and J82 human bladder carcinoma cells. Amine arrays were fabricated by immobilizing 3-aminopropyltrimethoxysilane on glass slides. The assay was specific and highly reproducible. The average coefficient of variation betweens spots was 2.6% (n = 3 arrays), and the average correlation coefficients between arrays and between arrays/reactions were 0.998 and 0.976, respectively (n = 3 arrays). The assay was successfully applied to detect changes in TG activity induced by maitotoxin and to analyze inhibition of the TG activation with cystamine and monodansyl cadaverine. In addition, the assay demonstrated that intracellular reactive oxygen species regulate the maitotoxin-induced activation of TG. Thus, the array-based in situ TG activity assay constitutes a rapid and high-throughput approach to investigating the roles of TGs in cell signaling. 相似文献