Electrophoretic assay of protein fractions,non-specific esterase fractions and acid phosphatase fractions in extracts from leaves and apple blossoms

Electrophoreograms of protein extracts from leaves of the variety Lord Lambourne, detached at the flowering period toward the end of April, differed quantitatively from electrophoreograms of protein extracts from leaves detached from shoots at the end of May and June. Electrophoreograms of protein extracts of leaves from juvenile seedlings, detached at the end of May and June, did not differ from electrophoreograms of protein extracts of leaves from shoots of the variety Lord Lambourne detached at the same period. Electrophoretic pattern of protein fractions and the fractions of non-specific esterase from leaves differed from the corresponding electrophoretic pattern of flowers quantitatively and in some areas even qualitatively; the pattern of acid phosphatase differed quantitatively. The results obtained are discussed in relation to organogenesis.     

Protein and glycoprotein composition of subsynaptosomal fractions

Highly enriched fractions of synaptic junctional complexes, synaptic vesicles, and coated vesicles were isolated from rat forebrains and compared, along with synaptosomal plasma membrane and its nonjunctional components, by discontinuous sodium dodecylsulfate-polyacrylamide gel electrophoresis. When stained for proteins with Coomassie blue, the gels all contained the same protein bands, only in different relative amounts. Mixing experiments revealed no additional bands. However, gel patterns of each fraction were not only quantitatively but also qualitatively different when stained for carbohydrate. These observations suggested that some of the protein bands may vary in their degree of glycosylation among the various synaptic fractions. Within the limits of resolution of the methods used here, these results are consistent with the morphological process of synaptic vesicle exocytosis and recycling but suggest the possibility of a reversible modification of certain membrane glycoproteins as they pass through the various membrane compartments.     

Immunogenic fractions of Cryptococcus neoformans

Cryptococcus neoformans cell and culture supernatant extracts were fractionated by ion exchange and gel filtration column chromatography. Various fractions were used to immunize mice, and to assess the release of migration inhibition factor, delayed type hypersensitivity, and protective immunity after challenge with C. neoformans. Results suggest that the C. neoformans fractions, which protect mice, contain a high molecular weight, predominantly carbohydrate antigen that can be distinguished from the capsular polysaccharide.     

Antioxidative and cytoprotective effects of Artemisia capillaris fractions

The antioxidant effects of Artemisia capillaris fractions against reactive oxygen species (ROS) were evaluated by measuring scavenging activities against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, superoxide (O_2(-)), hydroxyl (HO.) and nitric oxide (NO.) radical. Among five solvent fractions, ethyl acetate fraction showed the highest total polyphenol and total flavonoid contents as 648.75 and 89.09 microg/mg, respectively. Also, the ethyl acetate fraction showed the highest scavenging activity; the 50% inhibitory concentration (IC50, microg/mg) value for DPPH, O_2(-), HO. and NO. radical scavenging were 4.76, 31.54, 69.34 and 74.63, respectively. Additionally, the highest inhibition of rat liver microsomal lipid peroxidation was observed by ethyl acetate fraction. Except for free radical-mediated protein damage, ethyl acetate fraction showed the highest scavenging activity. The effect of Artemisia capillaris fractions on cell viability and DNA damage induced by H2O2 in Raw 264.7 cell were also evaluated by MTT and comet assay, respectively. The protective effect of ethyl acetate fraction, as indicated by cell viability increasing 71% and DNA breakage decreasing 51% as compared with H2O2-treated positive control. These results suggest that ethyl acetate fraction possess significant ROS scavenging and protective effect against oxidative DNA damage.