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41.
The ester groups of glycerophospholipids in tissue extracts can be cleaved in less than 10 min at room temperature if the lipids are extracted with hexane-isopropanol and the filtrate is treated with methanolic NaOH. The resulting mixture can be treated with aqueous Na-sulfate containing sulfuric acid and partitioned to remove the inorganic reagents and hydrophilic ester degradation products. When the procedure is applied to brain lipid extracts, the addition of alkali produces a second, lower phase that contains much of the hydroxycerebroside, virtually all of the sulfatide in the extract, and small amounts of other lipids. The sulfatide can be isolated from the lower phase by neutralizing it with HCl in aqueous methanol, and partitioning with chloroform to remove nonlipid components. The lower phase is evaporated to dryness and treated with periodic acid to convert the cerebroside to a less polar product. The lipids recovered from the reaction mixture are then fractionated with a Florisil column, which yields highly purified sulfatide. Starting with 300 g of pig brain one can obtain about 1.1 g of sulfatide in 4 working days, using conventional, compact equipment. Since the precipitation step is almost complete, and the procedure can be scaled down to very low levels, the method has promise for quantitation methods and isotopic studies of sulfatide metabolism. 相似文献
42.
43.
Data are presented on the cytokinin status of seeds and seed components, at different stages of development in Phaseolus coccineus L., as determined with the soybean callus growth bioassay: A change in cytokinin types according to developmental stage occurred: from biologically very active less polar types (zeatin=Z) at early stages to more polar types (zeatin glucoside=Z9G and zeatin riboside=Zr), with relatively low biological activity, at intermediate and late stages of seed development: When cytokinins were analyzed separately in embryos (embryo proper) and suspensors at two embryonic stages: heart-shaped (A) and middle cotyledonary embryos (stage B) respectively, it was found that: i) at stage A, the suspensor showed cytokinin activity at the level of Z, 2iPA (2-isopentenyladenosine) and Zr, whereas more polar cytokinins (Z9G, Zr) were present in the embryo; ii) at stage B, when the embryo seems to become autonomous for cytokinin supply, there was a relative abundance of active cytokinins (Z, 2iPA) in the embryo to which Z9G activity in the suspensor corresponded. It is concluded that the suspensor plays an essential role in embryogenesis by acting as a hormone source to the early embryo.Abbreviations GA
gibberellic acid
- 2iPA
2-isopentenyladenosine
- Stage A
heart-shaped embryo
- siage B
middle cotyledonary embryo
- Z
zeatin
- Z9G
zeatin glucoside
- Zr
Zeatin riboside 相似文献
44.
Yasuko Mizuno Atsushi Ichikawa Kenkichi Tomita 《Prostaglandins & other lipid mediators》1983,26(5):785-795
The effect of 7-fluoro proscyclilin (PGI2-F), a chemically stable analogue of prostacyclin, on cAMP accumulation in and [3H]PGE binding to mastocytoma P-815 cells was compared with those of the Na salt and methyl ester of prostacyclin (PGI2Na or PGI2Me), which are rapidly inactivated in aqueous solution or metabolized in the tissue.PGIF was as effective as PGI2Me, and slightly less effective than PGI2Na in stimulating cAMP accumulation in mastocytoma cells and rabbit platelets. PGI2F was also more stable than PGI2Me or PGI2Na, and retained its original cAMP elevating activity even after incubation with or without cells for 4 h at 37°C. Cells which had been exposed to PGI2F and then washed free of unbound reagent continued to produced cAMP for more than 3 h. PGI2F was also as effective as PGE1 or PGE2 in displacing [3H]PGE2 bound to the cells. Non-competitive inhibition by PGI2F or PGI2Me of [3H]PGE2 binding to the cells, with apparent Kis of 1.29 μM and 1.13 μM, respectively, indicates the presence of different receptors for PGE2 and for PGI2F or PGI2Me in mastocytoma P-815 cells. 相似文献
45.
Y Kameda 《The journal of histochemistry and cytochemistry》1992,40(4):541-553
A monoclonal antibody (MAb) that reacted only with thyroid C-cells was raised against cell suspensions from dog thyroid glands, to examine a glycoprotein secreted by C-cells. After chronically-induced hypercalcemia and administration of an anti-thyroid drug, reaction products for the antibody markedly decreased in C-cells, coinciding with alterations in calcitonin immunoreactivity. The antigen recognized by the MAb appears to be a secretory protein. The MAb reacted with C-cells from a wide variety of mammalian species, including rats, mice, hamsters, cattle, cats, rabbits, and monkeys. Furthermore, tumor cells of human medullary thyroid carcinoma, which is derived from C-cells, were immunoreactive to the MAb. Exceptionally, C-cells from guinea pigs and pigs were not stained with the MAb. No crossreactivity was observed in any of the dog tissues examined. Immunoblot analysis demonstrated that the MAb recognized a single prominent band at a molecular weight of approximately 79,000. The 79 KD band reacted with various digoxigenin-labeled lectins, including GNA, DSA, SNA, and MAA; it is a glycoprotein containing mannose, N-acetylglucosamine, and sialic acid. Dog thyroid C-cells were also densely stained with these lectins. The results indicate that thyroid C-cells synthesize and secrete a specific glycoprotein in addition to peptide hormones. 相似文献
46.
Occurrence of colloid-containing follicles and ciliated cysts in the hypophysial pars tuberalis from guinea pigs of various ages 总被引:2,自引:0,他引:2
Y Kameda 《The American journal of anatomy》1990,188(2):185-198
Colloid-containing follicles and ciliated cysts in the hypophysial pars tuberalis of guinea pigs at various ages ranging from 5 days to 36 months were examined by periodic acid-Schiff (PAS) reaction, immunohistochemistry, and electron microscopy. The follicles storing PAS-positive colloid were encountered in the pars tuberalis of all guinea pigs examined, although only a few were present in young animals. The follicles gradually increased in number with age. The largest number of follicles was found in the senile male group: 141.3 +/- 11.9, about 10 times the number in the 5-day-old male group. The follicles were scattered throughout the entire length of the pars tuberalis. Follicles with enlarged luminal cavities were concentrated in the ventral caudal region surrounding the infundibular stem and merges with the pars distalis. Three different types of follicles were found by electron microscopy: 1) those surrounded by nongranulated follicular cells that may correspond to the stellate-follicular cells in the pars distalis, 2) those surrounded by specific cells that were packed with vesicular inclusions, and 3) those surrounded by granulated cells that may be gonadotropes. In the follicles lined by non-granulated follicular cells, long, prominent microvilli and cytoplasmic processes protruding into the lumen and invaginations of colloid were often observed at the apical cell region. The follicles lined by the specific cells having numerous vesicles were localized only in the ventral caudal portion. The vesicles ranged from 200 to 700 nm in diameter, and the outer surface of their limiting membrane was partly studded with ribosomes. Gonadotropes immunoreactive to the luteinizing hormone (LH) and follicle-stimulating hormone (FSH) antisera were distributed in the guinea pig pars tuberalis. As well as the typical follicles described above, the follicles composed solely of granulated cells showed microvilli protruding into the cavities and junctional complexes at the apical lateral surface. They stored heterogeneous materials in the lumina. Some secretory granules gave the appearance of being discharged into the lumen. Ciliated cysts were frequently observed in the pars tuberalis; their incidence was 71.7%. The ciliated cysts were much larger than colloid-containing follicles. Cystic cavities were only partly filled with heterogeneous materials showing colloid-like, flocculent, and granular features. 相似文献
47.
Dr. Yoko Kameda 《Cell and tissue research》1982,225(3):693-697
Summary Small follicles composed solely of C cells were occasionally observed in large C cell groups of dog thyroid glands. The lumina of C-cell follicles were filled with, or contained peripheral depositions of PAS-positive amorphous material, which was similar in ultrastructural features to thyroglobulin-containing colloid in typical thyroid follicles. This indicates that C cells, in addition to secreting calcitonin, produce a glycoprotein that can be stored in the lumina of the follicles. 相似文献
48.
The product of the malE—lacZ gene fusion was reported to compete with some proteins including outer membrane lipoprotein in the protein translocation across the Echerichia coli membrane. The fusion product also inhibited colicin E1 export. Furthermore, globomycin, which accumulated prolipoprotein in the membrane, inhibited the translocation of colicin E1 in the wild-type cells, but not in lipoprotein-negative mutant cells. Since colicin E1 contains the internal signal-like sequence [Proc. Natl. Acad. Sci. USA (1982) 79, 2827–2831], these results suggest that colicin E1 is exported by the aid of this sequence at a common site for maltose-binding protein and lipoprotein translocation. 相似文献
49.
Protoplasts prepared from suspension-cultured Vinca rosea cellswere cultured for 5 days. The cell walls regenerated from theprotoplasts were mainly composed of glucans having 1,3- and1,4-linkages. To investigate the molecular species, these glucanswere separated into four fractions: EDTA (50 mM, pH 4.5)-soluble(fraction E), KOH (24%)- soluble but not precipitatable by neutralizationwith acetic acid (fraction K-S), KOH (24%)-soluble and precipitatableby neutralization with acetic acid (fraction K-P), and KOH (24%)-insoluble(fraction C). By means of sugar composition analysis, methylationanalysis, periodate oxidation and enzymatic digestion, the molecularspecies of the glucans contained in the regenerated cell wallswere deduced to be ß-1,4-glucan (cellulose) and ß-1,3-glucan.Fraction C was mainly composed of ß-1,4-glucan; ß-1,3-glucanwas mainly recovered in fraction K-P. The ß-l,3-glucanwas soluble in dilute alkali solution, but was only slightlysoluble in water. The ß-1,3-glucan had an essentiallyunbranched structure, and its weight average molecular weightestimated by gel permeation chromatography was 4.55.0x 104.
1 Present address: Division of Environmental Biology, NationalInstitute for Environmental Studies, Yatabe, Tsukuba, Ibaraki305, Japan (Received May 21, 1981; Accepted October 13, 1981) 相似文献
50.
Yoshikazu Kuroki Ichiro Matsui Yoshifumi Yamamoto Atsushi Ieshima 《Human genetics》1980,55(2):227-229
Summary Disomic and trisomic cells of a patient with Down syndrome mosaic were used to study the effect of the additional chromosome 21 against an identical genetic background. The frequency of Ag staining and the participation in satellite associations were determined for each pair of acrocentric chromosomes. The additional chromosome 21 of the trisomic cells and its homologues proved to be regularly Ag positive. Therefore the trisomic cells showed more Ag positive chromosomes and more satellite associations per cell than the diploid cells. Thus, no compensation for the additional rRNA-gene dose could be found in the cells of the trisomic line. 相似文献