Partial purification and characterization of polyphenol oxidase from banana (Musa sapientum L.) peel

Polyphenol oxidase (EC 1.10.3.1, o-diphenol: oxygen oxidoreductase, PPO) of banana (Musa sapientum L.) peel was partially purified about 460-fold with a recovery of 2.2% using dopamine as substrate. The enzyme showed a single peak on Toyopearl HW55-S chromatography. However, two bands were detected by staining with Coomassie brilliant blue on PAGE: one was very clear, and the other was faint. Molecular weight for purified PPO was estimated to be about 41 000 by gel filtration. The enzyme quickly oxidized dopamine, and its Km value (Michaelis constant) for dopamine was 3.9 mM. Optimum pH was 6.5 and the PPO activity was quite stable in the range of pH 5-11 for 48 h. The enzyme had an optimum temperature at 30 degrees C and was stable up to 60 degrees C after heat treatment for 30 min. The enzyme activity was strongly inhibited by sodium diethyldithiocarbamate, potassium cyanide, L-ascorbic acid, and cysteine at 1 mM. Under a low buffer capacity, the enzyme was also strongly inhibited by citric acid and acetic acid at 10 mM.     

A global map of human impact on marine ecosystems

The management and conservation of the world's oceans require synthesis of spatial data on the distribution and intensity of human activities and the overlap of their impacts on marine ecosystems. We developed an ecosystem-specific, multiscale spatial model to synthesize 17 global data sets of anthropogenic drivers of ecological change for 20 marine ecosystems. Our analysis indicates that no area is unaffected by human influence and that a large fraction (41%) is strongly affected by multiple drivers. However, large areas of relatively little human impact remain, particularly near the poles. The analytical process and resulting maps provide flexible tools for regional and global efforts to allocate conservation resources; to implement ecosystem-based management; and to inform marine spatial planning, education, and basic research.     

Characteristics of an aminopeptidase from Japanese cedar (Cryptomeria japonica) pollen

A peptidase from Japanese cedar pollen, Jc-peptidase, was clarified to preferentially hydrolyze an MCA substrate of Phe-MCA (L-phenylalanyl-4-methylcoumaryl-7-amide). This study examined substrate specificities of Jc-peptidase using oligopeptides. Jc-peptidase hydrolyzed Phe-Phe and Tyr-Phe effectively and hydrolyzed Leu-Phe, Met-Phe, and Arg-Phe moderately. Other substrates such as Ala-Phe, Asp-Phe, and Pro-Phe were not hydrolyzed with the peptidase. Results obtained with a series of aminoacyl-Phe peptides were compatible with the facts obtained for MCA substrates except for Arg-MCA. Effects of amino acid residues in the P1' position were also examined using Phe-amino acids. An N-terminal phenylalanine residue was actually released from bioactive peptides such as molluscan cardioexcitatory neuropeptide (FMRF-NH(2)). Because the activity was inhibited with Zn(2+) and EDTA, Jc-peptidase was inferred to belong to the metalloproteases. The N-terminal amino acid sequence was determined to be APIGVQLEIEENYVHMYNGF and an internal sequence to be EIFAATFNVDEETEA, but no homology with other proteins was found.     

光照强度对菠菜光合色素的影响

本丈以400、240、50μmol·m^-2·s^-1 3种不同光照强度处理菠菜,测定菠菜叶片中叶绿素a、叶绿素b、β-胡萝卜素和叶黄素含量的动态变化。结果表明：菠菜叶绿素a、叶绿素b的含量随着光照强度的增强而逐渐升高,且在高光强（4000μmol·m^-2·s^-1）下．植株叶片中2种色素含量随处理时间的延长呈逐渐升高的趋势,对照（2400μmol·m^-2·s^-1）植株含量在处理期间无明显变化．低光强（500μmol·m^-2．s^-1）下,2种色素含量呈降低的趋势,30d时各处理间差异显著;β-胡萝卜素含量随着光照强度的降低呈升高的趋势,30d时低光强处理植株含量显著高于对照和高光强处理植株;叶黄素含量：低光强〉高光强〉对照,20d和30d时．低光强处理植株含量显著高于高光强和对照植株。可以推论,高光强有利于菠菜叶绿素的合成,低光强有利于类胡萝卜素的形成。     

The state and fate of Himalayan glaciers

Himalayan glaciers are a focus of public and scientific debate. Prevailing uncertainties are of major concern because some projections of their future have serious implications for water resources. Most Himalayan glaciers are losing mass at rates similar to glaciers elsewhere, except for emerging indications of stability or mass gain in the Karakoram. A poor understanding of the processes affecting them, combined with the diversity of climatic conditions and the extremes of topographical relief within the region, makes projections speculative. Nevertheless, it is unlikely that dramatic changes in total runoff will occur soon, although continuing shrinkage outside the Karakoram will increase the seasonality of runoff, affect irrigation and hydropower, and alter hazards.     

Complex apocrine carcinoma with dominant myoepithelial proliferation in a dog

A rare case of complex apocrine carcinoma displaying dominant myoepithelial proliferation developed in the right leg subcutis of a 10-year-old male dog. The major cell population consisted of diffusely proliferating p63-expressing neoplastic cells that were largely myoepithelial in origin co-expressing α-smooth muscle actin. A small portion of the cell population consisted of concomitant basal epithelial cells lacking α-smooth muscle actin expression. The minor population consisted of p63-negative apocrine gland cells that expressed cytokeratin 8. The myoepithelial cell population showed a rather stronger proliferation activity than did the apocrine epithelial population. Thus, this tumor might have been derived from basal epithelial cells characterized by more predominant myoepithelial differentiation than luminal apocrine epithelial differentiation.     

Particle Size Effects on the Quality of Flour Tortillas Enriched with Whole Grain Waxy Barley

Wheat tortillas were enriched with whole barley flour (WBF) of different particle sizes including 237 μm (regular [R]), 131 μm (intermediate [IM]), and 68 μm (microground [MG]). Topographical and fluorescent microstructure images of flours, doughs, and tortillas were examined. Flours and tortillas were analyzed for color, protein, ash, starch, moisture, and β‐glucan content. Farinograph testing was conducted on the flour blends. Water activity and texture analyses of tortillas were conducted. A 9‐point hedonic scale was used by 95 untrained panelists to evaluate tortilla appearance, color, flavor, texture, and overall acceptability. Two commercial products (CP) were included in some analyses. As WBF particle size decreased, color was lighter; protein, moisture content and mixing stability decreased; ash, starch content, water absorption and farinograph peak time increased; and β‐glucan content was constant. WBF tortillas were darker than the control (C), while IM and MG tortillas had lower peak forces than C. No flavor differences were reported among C, R, and MG tortillas but higher scores were given to both CP in all attributes tested. Tortillas made with the largest WBF particle size (R) were the most similar in protein content, texture and flavor when compared with C tortillas made with refined bread flour.     

Isolation and genetic analysis of Japanese encephalitis virus from a diseased horse in Japan

Japanese encephalitis (JE) developed in an unvaccinated half-bred horse kept in Tottori Prefecture, Japan. The animal showed ataxia with pyrexia and low appetite, and ultimately died. A viral strain was isolated from the cerebrum of the horse and was identified as JE virus (JEV) by RT-PCR using JEV specific primers. The isolated JEV was classified into genotype I by nucleotide sequence analysis of the viral envelope gene. We believe that this is the first report of the genotype I strain being isolated from a horse.     

Efficient digestion and structural characteristics of cell walls of coffee beans

Screening of effective food-processing cellulase for digestion of cell walls of coffee beans was carried out, and the cellulase from Trichoderma sp. was selected. The digestion of the cell walls of green and roasted coffee beans was carried out by sequential procedures of alkali boiling (0.1 M Na2CO3 buffer, pH 10, and 0.1 M NaOH), cellulase digestion, autoclaving with 0.1 M NaOH, and cellulase redigestion. The total digestion yields were >95 and >96%, respectively. The cell walls became thin, and the final residues of the cell walls were easily broken into small pieces. The neutral sugar analysis of the digestion or the extract and the residues and the microscopy observations with staining with toluidine blue O, Yariv reagent, and calcofluor for the residue in each step were investigated. Four structures, the galactomannan-cellulose (center part), the membrane of the arabinogalactan protein, the cellulose-rich galactomannan layer, and the arabinogalactan protein-rich layers (outer part), were found in the cell walls.     

Regulatory effect of amino acids on the pasting behavior of potato starch is attributable to its binding to the starch chain

The binding of an amino acid, glycine (Gly), alanine (Ala), epsilon-aminocaproic acid (-AC), monosodium glutamate (GluNa), or lysine (Lys), to starch was examined by a biomolecular interaction analyzer (IAsys). A starch sample (ATS) hydrolyzed to an extent of 1% hydrolysis rate with 15% sulfuric acid was used as a model starch for the binding examination. The reducing end of ATS was oxidized by the Somogyi reagent, and the conversion of the reducing end to the carboxyl group of ATS was confirmed by a carboxylic acid fluorescence labeling reagent. The oxidized ATS was immobilized to the amino group of a sensor cuvette by using water-soluble carbodiimide and N-hydroxysuccinimide through an amide bond. The IAsys examination showed that Gly, Ala, and epsilon-AC scarcely bound to the immobilized starch chains but that GluNa and Lys favorably bound with their increasing concentrations. The relative binding index (RBI) of each amino acid was defined by the ratio of the slope of the linear regression equation between the binding response and the concentration for each amino acid to that for Gly. Because the relationships between the RBI and the pasting characteristics (pasting temperature, peak viscosity, breakdown, and swelling index) could each be expressed by a linear regression equation with a high correlation coefficient, it is concluded that the regulation of the pasting behavior of starch with an amino acid is caused by binding of the amino acid to the starch chains.