beta-Lactoglobulin A with N-ethylmaleimide-modified sulfhydryl residue, polymerized through intermolecular disulfide bridge on heating in the presence of dithiothreitol

Roles of sulfhydryl groups on thermal aggregation of beta-lactoglobulin A (betaLG A) at pH 7.5 were investigated. It is known that betaLG A modified at Cys(121) with N-ethylmaleimide (NEM-betaLG A) does not form an aggregate by heating and that dithiothreitol (DTT) reduces cystine residues and induces the intermolecular sulfhydryl/disulfide interchange reaction and/or oxidation. NEM-betaLG A was heated in the presence of DTT. The molecules were linked together with an intermolecular disulfide bridge, and the polymer formed increased with increase in DTT concentration. The largest portion of polymer was formed when DTT was added at around the same molar concentration as that of NEM-betaLG A. Then, polymer formation decreased with further increase in DTT concentration. The results suggest that sulfhydryl/disulfide residues other than Cys(121), generated from cysteine residues, can induce intermolecular sulfhydryl/disulfide interchange reactions to polymer and that thiol compounds, for example, added DTT, are capable of starting such reactions.     

Use of Jeffries acid oxalate treatment in particle-size analyses of Ando soils

Use of Jeffries acid-oxalate treatment in particle-size analyses of Ando soils for breaking up the sand- and silt-size aggregates bound by allophane, allophane-like constituents and hydrous iron oxides was studied. The analyses by this method and those by a method using sonic-wave oscillation were compared for nine Ando soil samples having different clay-mineral compositions and organic-matter contents. There were minor differences in the sand contents between the two analyses, but the clay contents were higher and the silt contents were lower after Jeffries acid oxalate treatment.

Characterization of the silt-size separates by an X-ray powder method, water vapor adsorption and electron microscopy indicated that the lower silt contents after Jeffries acid oxalate treatment were mainly due to dissolution of allophane and allophane-like constituents, whereas the silt-size separates after sonic-wave treatment still contained these mineral constituents. The proposed procedure for particle-size analysis was also useful for two ferruginous soils representing the Hydrandept and Torrox.     

Identification of whale species by thin-layer isoelectric focusing of sarcoplasmic proteins.

Thin-layer isoelectric focusing was applied to the identification of whale (Cetacea) species by using water-soluble sarcoplasmic proteins of skeletal muscles. Twenty-eight samples consisting of 4 species (10 samples) of baleen whales (Mysticeti) and 8 species (18 samples) of toothed whales (Odontoceti) were analyzed. Each sample (approximately 1 g) was electrophoresed with Ampholine PAGplate, pH 3.5-9.5. The electrophoretic profiles were species-specific on the 4 toothed whale species that did not have a marked intra-species difference, and all 4 baleen whale species. However, the profiles were not specific on the 4 other dolphin species, even though they were discriminable from the other 4 toothed whale species. Numerical values of pIs and relative peak heights were obtained by densitometric analysis of the isoelectro-focused protein bands. The bands were also species-specific for the 8 toothed whale species mentioned. The values may be applicable to species identification without the need for a standard sample, which may not be readily obtainable. Experiments on test samples of minke and sel whales showed that bloodletting with ice water made the densities of isoelectro-focused bands thinner, although species identification was still possible by using the inside part of muscles. Heat treatment at below 60 degrees C for 10 min caused little denaturation; at higher temperatures the protein bands were diminished in a temperature-dependent fashion. Therefore, the present isoelectric focusing analysis should be applicable to small samples of whale meat, excluding several species of dolphins.     

Phenolics composition and antioxidant activity of sweet basil (Ocimum basilicum L.)

The antioxidant activity of a methanolic extract of Ocimum basilicum L. (sweet basil) was examined using different in vitro assay model systems. The crude extract was fractionated on a Sephadex LH-20 column, and six fractions were identified. The DPPH scavenging assay system and the oxidation of the soy phosphotidylcholin liposome model system were used to evaluate the antioxidant activity of each fraction. Fraction IV showed the strongest activity followed by fractions V and VI. Phenolic compounds responsible for the antioxidative activity of the fractions were characterized by atmospheric pressure chemical ionization liquid chromatography-mass spectrometry. The major antioxidant compound in fraction IV was confirmed as rosmarinic acid by (1)H NMR and characteristic fragmentations in the mass spectrum. Moreover, the native of antioxidant activity of rosmarinic acid in the liposome system was examined. The results showed that one rosmarinic acid can capture 1.52 radicals, and furthermore, the existence of a synergistic effect between alpha-tocopherol and rosmarinic acid was revealed.     

Characterization of epoxydecenal isomers as potent odorants in black tea (Dimbula) infusion

In a black tea (Dimbula) infusion, the potent "sweet and/or juicy" odorants were identified as the cis- and trans-4,5-epoxy-(E)-2-decenals by comparison of their gas chromatography retention indices, mass spectra, and odor quality to those of the actual synthetic compounds. Of the two odorants, cis-4,5-epoxy-(E)-2-decenal has been identified for the first time in the black tea. On the basis of the aroma extract dilution analysis on the flavor distillate obtained using the solvent-assisted flavor evaporation technique from the black tea infusion, these isomers showed higher flavor dilution (FD) factors. The FD factors and concentrations of these odorants in the black tea infusion were observed to be much higher than those from Japanese green tea. In addition, the model studies showed that these odorants were generated from linoleic acid and its hydroperoxides by heating, but the generated amounts of these odorants from linoleic acid were much less than those of its hydroperoxides. It can be assumed from these results that the withering and fermentation, which are characteristic processes during the manufacturing of the black tea, which includes the enzymatic reaction such as lipoxygenase, is one of the most important factors for the formation of the epoxydecenal isomers.     

Seroepidemiological evidence for the possible presence of Babesia (Theileria) equi and Babesia caballi infections in donkeys in western Xinjiang, China

The prevalence of Babesia (Theileria) equi and B. caballi infections in donkeys in western Xinjiang China was investigated. In total, 93 serum samples were randomly taken from donkeys in the Kashi and Ili areas, and examined for B. equi and B. caballi infections by enzyme-linked immunosorbent assays using recombinant antigens. Of the 93 samples, 9 (9.6%) and 36 (38.7%) samples were positive for B. equi infection and B. caballi infection, respectively. In addition, 2 (2.2%) samples were positive for both B. equi and B. caballi infections. These results indicate that equine babesiosis might be extensively prevalent in donkeys in western Xinjiang.