Molecular characterization of avian polyomavirus isolated from psittacine birds based on the whole genome sequence analysis

Seven avian polyomaviruses (APVs) were isolated from seven psittacine birds of four species. Their whole genome sequences were genetically analyzed. Comparing with the sequence of BFDV1 strain, nucleotide substitutions in the sequences of seven APV isolates were found at 63 loci and a high level of conservation of amino acid sequence in each viral protein (VP1, VP2, VP3, VP4, and t/T antigen) was predicted. An A-to-T nucleotide substitution was observed in non-control region of all seven APV sequences in comparison with BFDV1 strain. Two C-to-T nucleotide substitutions were also detected in non-coding regions of one isolate. A phylogenetic analysis of the whole genome sequences indicated that the sequences from the same species of bird were closely related. APV has been reported to have distinct tropism for cell cultures of various avian species. The present study indicated that a single amino acid substitution at position 221 in VP2 was essential for propagating in chicken embryonic fibroblast culture and this substitution was promoted by propagation on budgerigar embryonic fibroblast culture. For two isolates, three serial amino acids appeared to be deleted in VP4. However, this deletion had little effect on virus propagation.     

Tyrosinase inhibitor from black rice bran

The inhibitor of tyrosinase activity in black rice bran was investigated. The methanol extract from black rice bran was re-extracted with hexane, chloroform, ethyl acetate, or water. The ethyl acetate extract had the most potent inhibition against tyrosinase activity by 80.5% at a concentration of 0.4 mg/mL. Inhibitory compound in the ethyl acetate fraction was isolated by silica gel column chromatography, and identified as protocatechuic acid methyl ester (compound 1) by GC, GC-MS, IR, and 1H and 13C NMR spectroscopy. Compound 1 inhibited 75.4% of tyrosinase activity at a concentration of 0.50 micromol/mL. ID(50) (50% inhibition dose) value of compound 1 was 0.28 micromol/mL. To study the structure-activity relationship, protocatechuic acid (2), vanillic acid (3), vanillic acid methyl ester (4), isovanillic acid (5), isovanillic acid methyl ester (6), veratric acid (7), and veratric acid methyl ester (8) were also assayed.     

Enlargement of the variances in amount and composition of anthocyanin pigments in sweetpotato storage roots and their effect on the differences in DPPH radical-scavenging activity

To investigate the effect of differences in anthocyanin composition on radical-scavenging activity in the purple-fleshed sweetpotato storage root, 124 sweetpotato clones with various compositions of anthocyanin were produced by crosses among superior sweetpotato genetic resources, and were analyzed for anthocyanin composition and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity (RSA). The highest content of anthocyanin among the clones was twofold higher than the content of cv. Ayamurasaki, which has been released for use in industrial pigment production in Japan. The range of variance in anthocyanin composition was very wide, as the ratio of peak areas of cyanidin-based anthocyanins to total anthocyanin peak areas ranged from 85.7% to 6.0%. The relationship between anthocyanin content and RSA was positively correlated. Correlation coefficients between RSA and the HPLC peak areas of cyanidin-based anthocyanins ranged from 0.606 to 0.687, whereas those of peonidin-based anthocyanins were 0.124–0.271. These results clearly demonstrated that cyanidin-based anthocyanins are closely related to DPPH RSA in sweetpotato storage roots.     

Physiological changes during feeding and rumination in cows

In this study, we investigated the physiological changes in cattle during feeding and rumination. We collected blood samples every 5 min by using an automated blood sampling system and simultaneously recorded feeding, ruminating, and other behaviors using a video camera. Plasma non‐esterified fatty acid (NEFA) concentrations continuously decreased during feeding and decreased temporarily during rumination. Plasma glucose concentration continuously decreased during feeding and remained stable during rumination. During feeding and rumination, there were no characteristic increases and subsequent decreases in plasma insulin and growth hormone (GH) concentrations, although insulin concentrations were positively correlated with glucose concentration. NEFA concentrations were not correlated with GH and insulin concentrations. In terms of chewing behavior, feeding and rumination are similar; therefore, the changes in metabolites such as NEFA might have been the same. Combination of behavioral observations and application of an automated blood sampling system could contribute to new findings on behavioral and physiological changes regarding the temporary decrease in plasma NEFA concentration during rumination in ruminants.     

Corneal Mineralization in Wistar Hannover Rats

We have recently started using Wistar Hannover rats in Japan and are now collecting background data. We have been frequently observing corneal mineralization in Wistar Hannover rats of both the RccHan^TM:WIST and Crl:WI (Han) strains. In this study, details of corneal mineralization in Wistar Hannover rats were histopathologically and ultrastructurally investigated. According to the results, Wistar Hannover rats had a much higher incidence of corneal mineralization compared with Sprague-Dawley rats. The incidence of corneal mineralization was higher in males than females. According to the histological examination, mineral deposits were positive for calcium by von Kossa’s method. Furthermore, in response to mineralization, keratocytes probably become active to play an important role against the mineralized substance.     

L‐carnitine improves hydrogen peroxide‐induced impairment of nuclear maturation in porcine oocytes

We investigated the effect of oxidative stress induced by hydrogen peroxide (H₂O₂) on lipid peroxide (LPO) level and nuclear maturation in porcine oocytes cultured with or without cumulus cells. After 22 h of pre‐culture, oocytes with attached cumulus cells (COC group) or denuded oocytes (DO group) were cultured with H₂O₂, and intra‐oocyte H₂O₂ and LPO levels were quantitatively analyzed using immunofluorescence. This is the first report evaluating LPO levels in porcine oocytes. After H₂O₂ supplementation, the DO group showed severe accumulation of H₂O₂ and LPO in the oocytes. Similarly, while inhibition of progression of nuclear maturation was observed in both groups, the effect was more severe in the DO group. These results demonstrate that cumulus cells reduce the accumulation of H₂O₂ stress in oocytes. Furthermore, we attempted to reduce the oxidative stress by H₂O₂ with L‐carnitine, a H₂O₂ scavenger. L‐carnitine decreased H₂O₂ and LPO levels in the oocytes in both groups, and improvement in the progression of impaired nuclear maturation was observed. These effects were different by the presence of cumulus cells. Our results provide that L‐carnitine is useful for alleviating H₂O₂‐induced oxidative stress by reducing LPO levels and improving the progression of nuclear maturation.     

Malignant Mesothelioma in the Thoracic Cavity of a Crj:CD(SD) Rat Characterized by Round Hyalinous Stroma

Spontaneous malignant mesothelioma was found in a 104-week-old male Crj:CD(SD) rat. The tumor was scattered on the surface of the lung, heart, mediastinal pleura and thoracic wall and metastasized to the alveolar septa. Histopathologically, small flattened or cuboidal tumor cells proliferated with stroma, formed almost normal papillary structures and reacted positively to colloidal iron stain and immunohistochemical staining for mesothelin. Round hyalinous stromata were pronounced, which is a characteristic feature, and the possible reason for this is as follows; at first, a small amount of collagen fibers was formed in the center of the clusters of several tumor cells, and then the cell clusters expanded like balloons with an increase in the collagen fibers.     

Effect of image characteristics of edge-grain patterns on visual impressions

To demonstrate the psychophysical effect of edge-grain patterns on visual impressions received by human observers, printed images of the edge-grain patterns of 16 species were prepared as well-regulated visual stimuli, and impressions of the patterns on 32 subjects were evaluated using six categories. Image characteristics of the patterns were expressed numerically using contrast values that were derived by multi-resolution contrast analysis (MRCA) proposed by the authors, and relationships between the image characteristics and the visual impressions were examined. The difference in lightness between local areas with a certain size in the image was used as the contrast value, which could be used to detect the change in lightness due to the particular feature in the edge-grain pattern. While some impressions were almost independent of the contrast values, other impressions showed highly positive or negative linear relationships to the contrast values, derived at the optimal filter size in MRCA. Moreover, these optimal sizes were rarely affected by color information. The contrast value proposed in this paper would be a good numerical index that could be used not only to describe image characteristics of the edge-grain patterns but also to estimate some of their visual impressions.