Viral RNA extraction using an automatic nucleic acid extractor with magnetic particles and genetic characterization of bovine viral diarrhea virus in Tokachi Province,Japan, in 2016–2017

In this study, the viral genome extraction performance of automatic nucleic acid extractors and manual nucleic acid extraction kits was compared. We showed that compared with manual kits, the automatic extractors showed superior genome extraction performance using bovine viral diarrhea virus (BVDV) genome-positive cattle sera and bovine coronavirus/infectious bovine rhinotracheitis virus-spiked cattle nasal swabs. In addition, the subgenotyping of BVDV strains detected in Tokachi Province in Japan during 2016–2017 was performed. Results showed that most of these BVDV strains belonged to subgenotype 1b, while few strains belonged to subgenotypes 1a and 2a. This study showed the high applicability of automatic nucleic acid extractors in extracting multiple viral genomes and the dominant subgenotype of BVDV in Tokachi.     

Lipid-rich bovine serum albumin improves the viability and hatching ability of porcine blastocysts produced in vitro

The effects of lipid-rich bovine serum albumin (LR-BSA) on the development of porcine blastocysts producedin vitro were examined. Addition of 0.5 to 5 mg/ml LR-BSA to porcine blastocyst medium(PBM) from Day 5 (Day 0 = in vitro fertilization) significantly increased the hatching ratesof blastocysts on Day 7 and the total cell numbers in Day-7 blastocysts. When Day-5 blastocysts were culturedwith PBM alone, PBM containing LR-BSA, recombinant human serum albumin or fatty acid-free BSA, addition ofLR-BSA significantly enhanced hatching rates and the cell number in blastocysts that survived compared withother treatments. The diameter, ATP content and numbers of both inner cell mass and total cells in Day-6 andDay-7 blastocysts cultured with PBM containing LR-BSA were significantly higher than in blastocysts culturedwith PBM alone, whereas LR-BSA had no effect on mitochondrial membrane potential. The mRNA levels of enzymesinvolved in fatty acid metabolism and β-oxidation (ACSL1, ACSL3,CPT1, CPT2 and KAT) in Day-7 blastocysts weresignificantly upregulated by the addition of LR-BSA. The results indicated that LR-BSA enhanced hatchingability and quality of porcine blastocysts produced in vitro, as determined by ATP content,blastocyst diameter and expression levels of the specific genes, suggesting that the stimulatory effects ofLR-BSA arise from lipids bound to albumin.     

Background level of indium and gallium in soil with special reference to the pollution of the soils from zinc and lead smelters

The mean and range of indium content in unpolluted soils (Humic Andosols, Eutric Fluvisols, and Mollic Gleysols) was found to be 0.037 (0.016–0.078) μg/g dry wt. In soils polluted by common heavy metals, elevated values of indium up to 1.92 μg/g dry wt were observed together with an increase of cadmium, zinc, and lead. In contrast to indium, gallium content in polluted soils was nearly the same as that in unpolluted soils throughout the profiles. The mean and range of gallium content in all unpolluted and polluted samples was 14.2 (12.8–16.3) μg/g dry wt. The indium and gallium contents in 4 Canadian reference soils were also analyzed.     

生物质资源高压连续输送的研究

建立了近中试规模的生物质资源高压连续输送装置，通过将生物质资源与水混成浆料的方法实现了生物质资源的高压连续输送、浆料的固含量可达10％（wt)以上，压力可达40MPa，最大浆料供应量为2．4L／h。     

Study on the glass transition for several processed fish muscles and its protein fractions using differential scanning calorimetry

ABSTRACT:   The glass transition behavior of processed fish muscles (bonito, tuna, mackerel, sea bream, cod) and its muscle protein fractions (sarcoplasmic and myofibrillar proteins) were studied using differential scanning calorimetry. Each dried processed fish muscle and the extracted protein fractions showed clear glass transition phenomenon. The T _g values of muscles and myofibrillar proteins from red muscle fishes tended to be lower than those from white muscle fishes though there was no difference on T _g of sarcoplasmic proteins. The T _g value of whole muscle was considerably lower than that of extracted protein fractions because of the plasticizing effects of low molecular weight materials contained in the muscle.     

Ligand-binding characteristics of feline insulin-binding immunoglobulin G

Polyclonal immunoglobulin (Ig) G autoantibodies against insulin have been identified in sera of healthy cats. We purified and fractionated insulin-binding IgGs from cat sera by affinity chromatography and analyzed affinity of insulin-binding IgGs for insulin and their epitopes. Following the passing of fraction A, which did not bind to insulin, insulin-binding IgGs were eluted into two fractions, B and C, by affinity chromatography using a column fixed with bovine insulin. Dissociation constant (KD) values between insulin-binding IgGs and insulin, determined by surface plasmon resonance analysis (Biacore™system), were 1.64e⁻⁴ M for fraction B (low affinity IgGs) and 2e⁻⁵ M for fraction C (high affinity IgGs). Epitope analysis was conducted using 16 peptide fragments synthesized in concord with the amino acid sequence of feline insulin by an enzyme-linked immunosorbent assay. Fractions B and C showed higher absorbance (affinity) of the peptide fragment of 10 amino acid residues at the carboxyl-terminal of the B chain (peptide No. 19), followed by peptide fragments of 6 to 15 amino acid residues of the B chain (peptide No. 8). Fraction C showed a higher absorbance to 7 to 16 amino acid residues of the B chain (peptide No. 5) compared with the absorbance of fraction B. Polyclonal insulin-binding IgGs may form a macromolecule complex with insulin through the multiple affinity sites of IgG molecules. Feline insulin-binding IgGs are multifocal and may be composed of multiple IgG components and insulin.     

Daily incremental lines in sika deer (Cervus nippon) dentine

This work was designed to observe the dentine incremental lines of the sika deer (Cervus nippon) fawns and to investigate their periodicity using the chronological labeling method with fluorochromes. The incremental lines were observed in decalcified specimens stained by Bodian's silver technique, and the fluorescence-labeled lines were observed in undecalcified and ground specimens. In the silver stained specimens, there were two types of lines, deeply stained thick lines and faintly stained minute regular incremental lines. The intervals and staining intensities of the deeply stained thick lines were very similar to those of the fluorescence-labeled lines in the ground specimens obtained from the same tooth, and hence, it appeared that the both lines were identical. The number of minute incremental lines between the deeply stained thick lines was the same as that of days between the time when each fluorescent labeling injection was made. Therefore, it seemed that each minute incremental line was formed each day. The possibility of age estimation in days using diurnal dentine increments was discussed.