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51.
High-affinity binding of [3H]folate to supernatant from homogenized human leukocytes containing large amounts of binding protein displayed apparent positive cooperativity. The DEAE-Sepharose® CL-6B chromatographic profile of the supernatant at pH 6.3 contained a major peak of folate binding (Mr approx. 25 000) in the front effluent and a smaller more acidic peak (Mr approx. 25 000) that emerged after a rise in NaCl from 30 mmol/l to 1 mol/l. Triton X-100 solubilized ceil sediment from the leukocyte homogenate contained some high-affinity folate binding activity (Mr approx 25 000), typically 5–10% of the total binding activity. 相似文献
52.
53.
Bård Smedsrød Håkan Pertoft Gösta Eggertsen Christer Sundström 《Cell and tissue research》1985,241(3):639-649
Summary This paper presents a study on the structure and function of Kupffer cells (KC) and liver endothelial cells (LEC) isolated by a simple and rapid technique involving 1) perfusion of the liver with collagenase; 2) cell separation by means of density centrifugation in Percoll; and 3) cell culture, taking advantage of the fact that KC and LEC differ in their preferences for growth substrate. The KC, which attach and spread under serum-free conditions on surfaces of glass or plastic during the first 15 min in culture exhibit a typical macrophage-like morphology including membrane ruffling and a heterogenous content of vacuoles. Moreover, these cells express (a) Fc receptors (FcR) for binding and phagocytosis of erythrocytes covered with immune globulin G (E-IgG), and (b) complement receptors (CR) for binding and serum dependent phagocytosis of erythrocytes covered with either human C3b or mouse inactivated C3b (iC3b). The cells also bind fluid phase fluoresceinated C3b. Approximately 30% of the KC express immune response-associated (Ia)-antigens.The LEC attach and spread on fibronectin coated surfaces, but not on glass or plastic surfaces, during the first two hours in culture with or without serum, and are morphologically distinct from KC. Cultured LEC are well spread out with no membrane ruffling and with numerous large vesicles surrounding the regularly shaped nucleus. These cells bind, but do not ingest E-IgG via the FcR, but no binding of fluid phase C3b or particle fixed C3b or iC3b can be observed. Incubation of LEC with fluorescein amine conjugates of ovalbumin or formaldehyde treated serum albumin, but not with fluoresceinated native serum albumin, results in accumulation of fluorescence specifically localized in the large perinuclear vesicles. Neither KC nor any other cell types tested have the ability to accumulate fluorescence upon incubation with these compounds. Iaantigens are not present on the LEC.Cytochemical demonstration of unspecific esterase, acid phosphatase, and peroxidase reveals different patterns and intensities of staining in KC as compared to LEC.Abbreviations Used
KC
Kupffer cells
-
LEC
Liver endothelial cells
-
C
Complement
-
C3b
Major fragment of C3 activation
-
iC3b
C3b that has been cleaved by factor I (C3b inactivator), present in serum
-
meC3b
C3b produced by treating purified human C3 with methyl amine
-
trC3b
C3b produced by treating purified human C3 with trypsin
-
CR
Complement receptors for C3b and iC3b
-
IgG
Immune globulin G
-
IgM
Immune globulin M
-
E
Erythrocytes
-
E-IgG
E covered with anti-E IgG
-
E-IgM E
covered with anti-E IgM
-
E-C3b(h)
E-IgM reacted with purified human C1, C4, oxidized C2 and C3 (E-IgMC14xyC2C3b)
-
E-iC3b(m)
E-IgM incubated with C5 deficient serum from AKR mice
-
FcR
Receptors for the Fc portion of IgG
-
FITC
Fluorescein isothiocyanate
-
FITC-meC3b
FITC conjugated to meC3b
-
FITC-trC3b
FITC conjugated to trC3b
-
FA
Fluorescein amine
-
FA-OA
Ovalbumin conjugated with FA
-
FA-SA
Serum albumin conjugated with FA
-
FA-FSA
Formaldehyde-treated serum albumin conjugated with FA
-
Ia
Immune response-associated AcE Acid unspecific esterase acting on alpha naphtyl acetate
-
NASDAE
Unspecific esterase acting on naphthol AS-D acetate
-
NASDCAE
Unspecific esterase acting on napthol AS-D chloroacetate 相似文献
54.
Jørn Erik Bjørndalen 《Plant Ecology》1985,59(1-3):211-224
Basiphilous pine forests and related birch forests are herb-and grass-rich forests on calcareous substrate. These forests are complex communities with floristic/ecological elements from different vegetation types occurring in a subtle micromosaic. These elements are e.g. species from acidophilous conifer forests, thermophilous forest-rim communities, calcareous shallow-soil and steppe communities, eutrophic wet meadows and fens, and in northern Fennoscandia also species from alpine Dryas heaths. Four associations are recognized in Fennoscandia: Convallario-Pinetum, Melico-Piceetum pinetosum, Peucedano-Pinetum and Epipacto atrorubentis-Betuletum. The main association is the Convallario-Pinetum, a widespread community in Fennoscandia and Estonia with a considerable floristic variation between the different regions. Examples of the floristic variation along west-east profiles and south-north profiles in Fennoscandia are presented. The basiphilous pine forest complex can be divided into a number of ecological types along the moisture and nutritional gradients. A further subdivision into geographical types (races) is presented.Nomenclature follows Lid (1974) for vascular plants, Nyholm (1954–1969) for musci and Dahl & Krog (1973) for lichens. 相似文献
55.
Steinar Solberg Tor Larsen Leif Jørgensen 《In vitro cellular & developmental biology. Plant》1985,21(11):612-616
Summary This study examined whether nonconfluent endothelial cell cultures reacted differently than confluent ones toward thrombin-stimulated
platelets or a heparinized salt solution. The adherence to the endothelial cell cultures of51Cr-labeled human platelets stimulated at different thrombin concentrations was studied. There was significantly higher adherence
of stimulated platelets to nonconfluent cultures compared with confluent ones. This was confirmed by scanning electron microscopy,
which also revealed a tendency for the platelets to adhere at the cell periphery. Electron microscopy also showed that thrombin-stimulated
platelets induced endothelial cell contraction. Part of the peripheral endothelial cell surface toward the bottom of the culture
dish was inverted, facing the lumen of the dish. This phenomenon was particularly seen in nonconfluent cultures. When51Cr-labeled endothelial cultures were incubated with a mildly injurious fluid as heparinized sodium acetate and 20% serum,
at 20° C for 30 min, the nonconfluent cultures showed significantly more cell detachment and release of51Cr than the confluent ones. We conclude that under the conditions of the present experiments there are differences in the
reactivity of confluent and nonconfluent endothelial cell cultures. These differences probably reflect biological dissimilarities.
In experiments where properties of cultured endothelium are studied, care should be taken that the degree of confluency is
standardized. 相似文献
56.
Jørgen Johansen Anna L. Kleinhaus 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1985,157(4):491-497
Properties of divalent cation potentials carried by either Sr2+ or Ca2+ ions in Na+-free, TEA-Ringer solution were characterized in identified neurons of two species of leeches (Macrobdella and Haementeria). In Macrobdella, the overshoot of the potentials varied logarithmically with [Sr2+]0 (28.5 mV per 10-fold change). The overshoot, Vmax, and duration of the potentials increased with increasing divalent cation concentration and saturated at about 20 to 30 mM [Sr2+]0. The Vmax, amplitude, and duration of the potentials were reversibly blocked by Co2+ and Mn2+. The block by Mn2+ could be well-fitted by a reverse Langmuir-curve with an apparent KI of 100 micromolar. The local anesthetic procaine also reversibly inhibited the Vmax and duration of the potentials. The inhibition was greater at alkaline pH suggesting that procaine blocks the calcium channel from inside the membrane. The identified leech neurons examined in Macrobdella varied considerably in their ability to sustain somatic divalent cation potentials. Stimulation of T cells and most motoneurons produced no or only weak potentials, whereas stimulation of Retzius, N, Nut, and AP cells evoked overshooting potentials of several seconds' duration. Stimulation of the ALG cell of Haementeria in normal Ringer solution evoked a slowly-rising, purely Ca2+-dependent potential of approximately 100 ms duration. This response was TTX-resistant, unaffected by complete removal of Na+ from the Ringer solution, and abolished by 1 mM Mn2+. The overshoot varied logarithmically with a slope of 28 mV/decade change in [Ca2+]0. 相似文献
57.
Centriole distribution during tripolar mitosis in Chinese hamster ovary cells 总被引:17,自引:10,他引:7
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During bipolar mitosis a pair of centrioles is distributed to each cell but the activities of the two centrioles within the pair are not equivalent. The parent is normally surrounded by a cloud of pericentriolar material that serves as a microtubule-organizing center. The daughter does not become associated with pericentriolar material until it becomes a parent in the next cell cycle (Rieder, C.L., and G. G. Borisy , 1982, Biol. Cell., 44:117-132). We asked whether the microtubule-organizing activity associated with a centriole was dependent on its becoming a parent. We induced multipolar mitosis in Chinese hamster ovary cells by treatment with 0.04 micrograms/ml colcemid for 4 h. After recovery from this colcemid block, the majority of cells divided into two, but 40% divided into three and 2% divided into four. The tripolar mitotic cells were examined by antitubulin immunofluorescence and by high voltage electron microscopy of serial thick (0.25-micron) sections. The electron microscope analysis showed that centriole number was conserved and that the centrioles were distributed among the three spindle poles, generally in a 2:1:1 or 2:2:0 pattern. The first pattern shows that centriole parenting is not prerequisite for association with pole function; the second pattern indicates that centrioles per se are not required at all. However, the frequency of midbody formation and successful division was higher when centrioles were present in the 2:1:1 pattern. We suggest that the centrioles may help the proper distribution and organization of the pericentriolar cloud, which is needed for the formation of a functional spindle pole. 相似文献
58.
Marianne Klint Anita Fridberger Alan Menge Jan Sllstrm Leif Plen 《Molecular reproduction and development》1987,17(2):173-190
Boar sperm glycoprotein fractions were isolated by Lens culinaris hemagglutinin affinity chromatography of detergent-solubilized ejaculated spermatozoa, followed by preparative sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. In order to develop methods for further investigations of the sperm proteins, we proceeded with two of the isolated glycoproteins. Antibodies were raised in female rabbits against each of the two sperm glycoproteins. By a combination of immunosorbent chromatography, using the antibodies obtained, and preparative SDS polyacrylamide gel electrophoresis, highly purified sperm proteins were isolated. The sperm proteins were immobilized on Sepharose gel columns and specific immunoglobulin Fab fragments were enriched by affinity chromatography. The specificity of the Fab fragments was ascertained by immunoprecipitation analysis. The Fab fragments were used in indirect immunofluorescence analysis to localize the corresponding antigens on the surface of boar spermatozoa. Both antigens were exclusively confined to the postacrosomal region. Immunohistochemical staining of boar testis sections revealed that both antigens are expressed from the spermatid stage. This technique also revealed that one of the antigens congregated at the Golgi complex-acrosome region during spermatogenesis. 相似文献
59.
Isopropylidenation of lactose with 2,2-dimethoxypropane in the presence ofp-toluenesulfonic acid gave two products, which were identified by1H- and13C-NMR as 2,35,63,4-tri-O-isopropylidenelactose dimethyl acetal (1) and its 6-O-(2-methoxy)-isopropyl derivative (2). These products were used for the synthesis of 2-O-methyllactose (7), 2,6-di-O-methyllactose (9) and 2-O-benzyllactose (13). 相似文献