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1.
Summary Three embedding media have been compared with respect to post-embedding immunolabeling of contractile and cytoskeletal antigens in aldehyde-fixed smooth muscle tissue: the methacrylate derivates lowicryl K4M (cured at –35 or 60°C) and LR White (cured at 0 or 60°C) and the water soluble resin, polyvinylalcohol (dried at 60°C). Measurements of intensity of labeling of ultrathin sections in the fluorescence microscope showed that five antigens (actin, myosin light chain, tropomyosin, filamin and vinculin) reacted more or less equally with their respective antibodies in all the embedding media, including those cured at 60°C. One antibody (anti-light meromyosin) reacted well only with polyvinylalcohol-embedded tissue. In contrast to the relative invariance of antibody reactivity between media clear differences in the preservation of ultrastructural integrity were observed. Embedding in polyvinylalcohol (dried at 60°C) and in Lowicryl (cured at –35°C) resulted in superior preservation as compared to Lowicryl or LR White cured at 60°C. Examples of uitrastructural immunocytochemistry with the antibodies against filamin and myosin light chain, using the immunogold staining procedure are presented: the sites of localization by these antibodies were the same with all the media tried. The relative merits of the different methods are discussed.Abbreviations EGTA
Ethyleneglycol-bis(-amino ethyl ether)N,N,N,N-tetra acetic acid
- PIPES
1,4-Piperazinediethanesulfonic acid
- LR
London Resin 相似文献
2.
Homologues of catalytic domains of Cellulomonas glucanases found in fungal and Bacillus glycosidases 总被引:1,自引:0,他引:1
Catherine A. West rzej Elzanowski Lai-Su Yeh Winona C. Barker 《FEMS microbiology letters》1989,59(1-2):167-172
We demonstrate homology between the catalytic domains of exoglucanase (1,4-beta-D-glucan cellobiohydrolase, EC 3.2.1.91) from Cellulomonas fimi and those of endoxylanases (1,4-beta-D-xylan xylanohydrolases, EC 3.2.1.8) from Bacillus sp. strain C-125 and the fungus Cryptococcus albidus; and between the catalytic domains of endoglucanase (1,4-(1,3:1,4)-beta-D-glucan 4-glucanohydrolase, EC 3.2.1.4) from Cellulomonas fimi and exoglucanase II from Trichoderma reesei. These five enzymes apparently evolved by reshuffling of two catalytic domains and several substrate-binding domains. 相似文献
3.
Alterations in cell membrane structure and expression of a membrane-associated protein after adaptation to osmotic stress 总被引:3,自引:0,他引:3
4.
We hypothesized that manganese deficient animals fed high vs moderate levels of polyunsaturated fat would either manifest
evidence of increased oxidative stress or would experience compensatory changes in antioxidant enzymes and/or shifts in manganese
utilization that result in decreased endogenous gut manganese losses. Rats (females in Study 1, males in Study 2,n = 8/treatment) were fed diets that contained 5 or 20% corn oil by weight and either 0.01 or 1.5 μmol manganese/g diet. In
study 2,54Mn complexed to albumin was injected into the portal vein to assess gut endogenous losses of manganese. The manganese deficient
rats:
Gut endogenous losses of manganese tended to account for a smaller proportion of absorbed manganese in rats fed high-fat diets;
otherwise fat intake had few effects on tissue manganese concentrations. 相似文献
1. | Had 30–50% lower liver, tibia, kidney, spleen, and pancreas manganese concentrations than manganese adequate rats; |
2. | Conserved manganese through ≈70-fold reductions in endogenous fecal losses of manganese; |
3. | Had lower heart manganese superoxide dismutase (MnSOD) activity; and |
4. | Experienced only two minor compensatory changes in the activity of copper-zinc superoxide dismutase (CuZnSOD) and catalase. |
5.
John V. Brigande rzej Wieraszko Mary D. Albert Grant W. Balkema Thomas N. Seyfried 《Journal of neurochemistry》1992,58(2):752-760
The E1 (epileptic) mouse is considered a model for complex partial seizures in humans. Seizures in E1 mice begin around 7-8 weeks of age and persist throughout life. To determine if astrocytic gliosis was present in adult seizing E1 mice, the distribution of glial fibrillary acidic protein (GFAP) was studied in the hippocampus using an antibody to GFAP. The mean number of GFAP-positive cells per square millimeter of hippocampus was approximately 15- to 40-fold higher in adult E1 mice than in nonseizing control C57BL/6J (B6) mice or in young nonseizing E1 mice. Relative GFAP concentration (expressed per milligram of total tissue protein) in hippocampus and cerebellum was estimated by densitometric scanning of peroxidase-stained western blots. GFAP concentration was 2.7-fold greater in hippocampus of adult seizing E1 mice than in the control B6 mice. No differences in GFAP content were detected between the strains in the cerebellum. Because gangliosides can serve as cell surface markers for changes in neuronal cytoarchitecture, they were analyzed to determine if the gliotic response in E1 mice was associated with changes in neural composition. Although the total ganglioside concentration of hippocampus, cerebral cortex, and cerebellum was similar in adult E1 and control B6 mice, a synaptic membrane enriched ganglioside, GD1a, was elevated in the adult E1 cerebral cortex and hippocampus. The findings indicate that E1 mice express a type of gliosis that is not accompanied by obvious neuronal loss. 相似文献
6.
7.
Amol A. Verma Tejasvi Hora Hae Young Jung Michael Fralick Sarah L. Malecki Lauren Lapointe-Shaw Adina Weinerman Terence Tang Janice L. Kwan Jessica J. Liu Shail Rawal Timothy C.Y. Chan Angela M. Cheung Laura C. Rosella Marzyeh Ghassemi Margaret Herridge Muhammad Mamdani Fahad Razak 《CMAJ》2021,193(23):E859
8.
9.
Anna Romanowska Ewa Katzenellenbogen Magorzata Kuakowska rzej Gamian Danuta Witkowska Marian Mulczyk Elbieta Romanowska 《FEMS microbiology letters》1988,47(3):151-155
Abstract Lipopolysaccharides (LPS) of 33 strains of Hafnia alvei were isolated and purified. LPS content of the dry bacterial mass ranged from 1.2 to 4.5%. All examined lipopolysaccharides contained glucose, glucosamine, heptose, 3-deoxy-octulosonic acid and often galactose. Rhamnose, mannose, galactosamine, mannosamine and unidentified amino sugars were found in some H. alvei strains. Sialic acid was present in LPS of one strain. d -3-Hydroxybutyryl groups also were identified in lipopolysaccharides of 5 strains of this genus.
SDS-PAGE of the lipopolysaccharides was presented in the paper. According to these results two core types exist in H. alvei . 相似文献
SDS-PAGE of the lipopolysaccharides was presented in the paper. According to these results two core types exist in H. alvei . 相似文献
10.
Vrljic M Garg J Bellmann A Wachi S Freudl R Malecki MJ Sahm H Kozina VJ Eggeling L Saier MH Eggeling L Saier MH 《Journal of molecular microbiology and biotechnology》1999,1(2):327-336
In Corynebacterium glutamicum the LysE carrier protein exhibits the unique function of exporting L-lysine. We here analyze the membrane topology of LysE, a protein of 236 amino acyl residues, using PhoA- and LacZ-fusions. The amino-terminal end of LysE is located in the cytoplasm whereas the carboxy-terminal end is found in the periplasm. Although 6 hydrophobic domains were identified based on hydropathy analyses, only five transmembrane spanning helices appear to be present. The additional hydrophobic segment may dip into the membrane or be surface localized. We show that LysE is a member of a family of proteins found, for example, in Escherichia coil, Bacillus subtilis, Mycobacterium tuberculosis and Helicobacter pylori. This family, which we have designated the LysE family, is distantly related to two additional protein families which we have designated the YahN and CadD families. These three families, the members of which exhibit similar sizes, hydropathy profiles, and sequence motifs comprise the LysE superfamily. Functionally characterized members of the LysE superfamily export L-lysine, cadmium and possibly quarternary amines. We suggest that LysE superfamily members will prove to catalyze export of a variety of biologically important solutes. 相似文献