Evaluation of chiral alpha-cyanoesters as general fluorescent substrates for screening enantioselective esterases

Esterases play a crucial role in industrial chemical synthesis, maintaining normal physiological metabolism and detoxifying exogenous ester-containing toxicants. To meet the rapidly increasing industrial need for all kinds of esterases, especially enantioselective esterases used to generate highly pure chiral compounds, general substrates are necessary for rapid screening, monitoring, purification, and characterization. In this study, general fluorescent substrates including phenolic derivatives and alpha-cyanoesters were evaluated for sensitivity in detecting esterases in buffer systems. Results with two different esterases and different incubation times suggested that the alpha-cyanoesters examined were significantly more sensitive at detecting esterases than the corresponding tested phenolic derivatives. More importantly, alpha-cyanoesters, containing a secondary alcohol, possess at least one chiral center; thus, they are tools to screen for enantioselective hydrolysis. Results indicated that the enantioselectivity of esterases toward general alpha-cyanoesters strongly depended on the esterase and the substrate, but the majority of esterases examined preferred S-isomers to their corresponding R-enantiomers. Most appealing was the very high enantioselectivity displayed in cytosolic esterases of the house fly. The potential utility of such esterases is discussed. In addition, the use of alpha-cyanoesters as chiral fluorescent substrates was demonstrated for monitoring in enantioselective esterases.     

Differential Expression of Two Cytosolic Ascorbate Peroxidases and Two Superoxide Dismutase Genes in Response to Abiotic Stress in Rice

Superoxide dismutase (SOD) and ascorbate peroxidase (APX) play central roles in the pathway for scavenging reactive oxygen species in plants, thereby contributing to the tolerance against abiotic stress. Here we report the responses of cytosolic SOD (cSOD; sodCc1 and sodCc2) and cytosolic APX (cAPX; OsAPX1 and OsAPX2) genes to oxidative and abiotic stress in rice. RNA blot analyses revealed that methyl viologen treatment caused a more prominent induction of cAPXs compared with cSODs, and hydrogen peroxide t...     

Molecular cloning and expression analysis of double-stranded RNA-dependent protein kinase (PKR) in rock bream (Oplegnathus fasciatus)

We have cloned and characterized rock bream double-stranded RNA-dependent protein kinase (PKR), which is a key component of type I IFN inducible innate immune system. Full-length rock bream PKR cDNA consists of 2115 bp ORF encoding 704 amino acids, 124 bp 5′ UTR, and 529 bp 3′ UTR. Conserved domain analysis revealed that rock bream PKR contained two tandem dsRBM and kinase domain consisted of 11 kinase sub-motifs, which are characteristics found in other PKRs. Rock bream PKR was constitutively expressed in the spleen of rock bream and, upon injection of poly I:C, up-regulated not later than 12 h post injection and returned to baseline level at 48 h post injection. Although the eIF2α kinase activity of rock bream PKR was not examined in the present study, dsRNA inducible nature of rock bream PKR implies the possible important role of this gene in innate immune system of rock bream as suggested in zebrafish and flounder.     

Small‐scale heterogeneity in the soil environment influences the distribution of lawn grass and weeds

The association of the micro‐scale distribution of six plant species in a lawn, including the lawn grass, Zoysia japonica, with the soil's environmental factors was studied in an urban park in Kyoto, Japan. The cover of each plant species was scored in July and August. The relative elevation, depth of waterlogging after a shower, soil water content, and soil hardness were measured in 144 quadrats in a 12 m × 12 m plot. The subsurface soil was sampled at every fourth quadrat and was analyzed for the percentage of fine earth and the levels of total carbon, total nitrogen, and available phosphate. Three correlation criteria, namely the Pearson's, Mantel, and partial Mantel correlations, showed considerable disagreement in indicating whether or not there was a significant correlation between each plant species and environmental variable. Although the reason for the disagreement was not always clear, some unimodal or inverted unimodal responses of a plant species against an environmental factor, which was not detectable by the Pearson's correlation coefficient, were indicated to be significant by the Mantel or partial Mantel tests. There were four plant–environment pairs that had a significant correlation in all three criteria: Juncus tenuis in August had a positive correlation with the soil water content in July and August and a negative correlation with soil hardness and Z. japonica showed a negative correlation with the water content in August. The results suggest that the small patches of lawn with poor drainage are the sites of invasion by J. tenuis.     

Heart rate variability after horse trekking in leading and following horses

Horse trekking (HT) is having a stroll on a horse along a walking trail in a forest, field, and/or sandy beach. Generally in HT, horses exercise in tandem line outside the riding facilities. Because the leading horse will be confronted with stressors in the forefront, we hypothesized that the leading horse shows higher stress responses than the following one. In order to verify the hypothesis, we compared short‐term stress responses between each position in six horses. Exercise consisted of 15 min of ground riding and 45 min of HT with walking and trotting. Heart rate variability was analyzed for 5 min at 30, 60, and 90 min after the exercising period. There was no significant difference in heart rate during exercise between leading and following positions. The high frequency / low frequency power band of heart rate variability, an index of sympathetic nervous activity, after exercise, tended to be higher in the leading position than following one (P < 0.1). The result in this study can suggest that the leading horse was in a higher stressed state than the following horse after HT.     

Changes in the ovary and the peripheral concentrations of sex hormones after the aspiration of follicular fluid from the spontaneous follicular cysts of dairy cows

The aim of the present study was to clarify the ovarian and hormonal dynamics after the aspiration of follicular fluid in cows with follicular cysts. Follicular fluid was aspirated from the follicular cysts and follicles that were fated to become cystic follicles and other coexisting normal follicles, respectively, in lactating cows (n = 3). After the aspiration procedure, new follicles developed and reached a diameter of 25 mm without ovulation within 13–19 days. The plasma concentrations of inhibin decreased and follicle-stimulating hormone increased rapidly after the aspiration procedure, and subsequently increased and decreased, respectively, as a new follicle grew. No luteal structures developed after the aspiration procedure, and the animals’ plasma progesterone levels remained low. The present study indicates that the cystic follicles are never luteinized by the aspiration of follicular fluid, and consequently, new follicular cysts are observed to repeatedly develop.     

Effectiveness of tori-lines for further reduction of incidental catch of seabirds in pelagic longline fisheries

To improve the performance of tori-lines (bird-scaring lines) in reducing incidental catch of large seabirds (albatrosses and giant petrels) in the pelagic longline fishery, we analyzed factors affecting the performance of tori-lines based on data collected by Japanese scientific observers in the southern bluefin tuna Thunnus maccoyii fishery from 2002 to 2005. We classified the variety of tori-lines by the main tori-line length and the streamer type. Two types of streamers were identified: type A, several long streamers made of nylon cord attached to the main tori-line by metal swivels; type B, many short streamers made of polypropylene packing bands braided into the main tori-line. In a model analysis, we found that the main tori-line length was an important factor affecting tori-line performance, and the two types of streamers had similar effectiveness. Since the light type B streamer has the advantage of practical feasibility, this type can be another option for use in tori-lines if used with a main tori-line of sufficient length.