Intranasal inoculation of chickens with encapsulated or nonencapsulated variants of Haemophilus paragallinarum: electron microscopic evaluation of the nasal mucosa

Chickens were inoculated intranasally with encapsulated or nonencapsulated strains of Haemophilus paragallinarum. The nasal mucosa of the chickens were examined, using scanning and transmission electron microscopy. In chickens given the encapsulated variant, marked loss of cilia and microvilli, infiltration of leukocytes, and deposition of a mucopurulent substance were seen on the surface of the nasal mucosa; the number of microvillous cells were markedly increased, and infiltration of mast cells into the lamina propria of the mucous membrane was observed. Numerous encapsulated organisms were found near cilia and on microvillous cells, and the capsule surrounding the organisms appeared to mediate attachment of the organisms to the cilia. In contrast, the nasal mucosa of the chickens given the nonencapsulated variants changed little morphologically. A few nonencapsulated organisms were found near cilia. Colonization of the nasal mucosa by encapsulated organisms probably was essential to induce the morphologic changes seen in the nasal mucosa.     

Generation of PDX‐1 mutant porcine blastocysts by introducing CRISPR/Cas9‐system into porcine zygotes via electroporation

Recently, we established the GEEP (“gene editing by electroporation of Cas9 protein”) method, in which the CRISPR/Cas9 system, consisting of a Cas9 protein and single guide RNA (sgRNA), is introduced into pig zygotes by electroporation and thus induces highly efficient targeted gene disruption. In this study, we examined the effects of sgRNA on the blastocyst formation of porcine embryos and evaluated their genome‐editing efficiency. To produce an animal model for diabetes, we targeted PDX‐1 (pancreas duodenum homeobox 1), a gene that is crucial for pancreas development during the fetal period and whose monoallelic disruption impairs insulin secretion. First, Cas9 protein with different sgRNAs that targeted distinct sites in the PDX‐1 exon 1 was introduced into in vitro‐fertilized zygotes by the GEEP method. Of the six sgRNAs tested, three sgRNAs (sgRNA1, 2, and 3) successfully modified PDX‐1 gene. The blastocyst formation rate of zygotes edited with sgRNA3 was significantly (p < 0.05) lower than that of control zygotes without the electroporation treatment. Our study indicates that the GEEP method can be successfully used to generate PDX‐1 mutant blastocysts, but the development and the efficiency of editing the genome of zygotes may be affected by the sgRNA used for CRISPR/Cas9 system.     

Dynamic properties of three types of wood-based composites

This study used a vibration test method to show that grain angles of face veneer have substantial effects on sound velocities and dynamic Youngs moduli of three types of wood-based composites. The sound velocity at 0° grain angle of face veneer was the highest, and it decreased with increasing grain angle in the range of 0° to 90°. This tendency was similar to that for dynamic Youngs modulus. The relationship between the grain angle of face veneer and the sound velocity of three types of wood-based composites can be expressed in the form of Hankinsons equation or a second-order parabolic equation. This study also showed that the application of orthotropic elasticity theory was valid for the three types of wood-based composites. The relationship between the grain angle of the face veneer and the Youngs modulus of three types of wood-based composites can be expressed in the form of the Jenkin equation, Hankinsons equation, or a second-order parabolic equation. Rule of mixture can also be used to predict the Youngs modulus of wood-based composite from the Youngs moduli of the two elements.Part of this study was presented at the 53rd Annual Meeting of the Japan Wood Research Society, Fukuoka, Japan, March 2003 and the 15th Annual Meeting of Chugoku Shikoku Branch of the Japan Wood Research Society, Higashi-Hiroshima, Japan, September 2003     

Temporal water deficit and wood formation in Cryptomeria japonica

Cell behavior in the cambium and developing xylem of 3-year-old Japanese cedar (Cryptomeria japonica D. Don.) trees, during and after an 11-day suspension of irrigation, was analyzed. Leaf xylem pressure potential and tangential strain of the stem surface were monitored throughout the experiment. Anatomical features and numbers of developing tracheids and cambial cells were observed in four trees, sampled on Days 0, 4, 8 and 11 after irrigation was suspended. Daytime xylem pressure potential decreased to -1.9 MPa on Day 7 and remained the same until irrigation was resumed on Day 11. The transverse dimensions of the tracheids, which began to form secondary walls, began to decrease on Day 4. The number of cells in the cambial zone and cell expansion zone decreased abruptly on Day 8. Tangentially aligned developing tracheids with collapsed cell walls were observed in samples harvested on Days 8 and 11. Secondary wall formation was recognized in these tracheids. After the resumption of irrigation, xylem pressure potential recovered rapidly to the same value as before the suspension of irrigation. Tangential strain increased within 30 min after the resumption of irrigation, and continued to increase until the onset of light the next day. Eighteen days after the resumption of irrigation, anatomical features of cells in the cambium and cell-expansion zone were similar to those observed before suspension of irrigation.     

Piezoelectric behavior of wood under combined compression and vibration stresses I: Relation between piezoelectric voltage and microscopic deformation of a Sitka spruce (Picea sitchensis Carr.)

This study investigated the relation between piezoelectric behavior and the deformation of trachieds in real time under combined compression and vibration stresses. Scanning electron microscope images were recorded directly into a video recorder. Two types of microscopic destruction were observed in the specimens. With the first type, although a small uprush around the boundary of the annual ring was observed, the specimens were broken only by shearing fracture in the 45° direction. With the second type, the specimens were finally broken by shearing fracture after repeated buckling. In these cases the piezoelectric voltage increased almost linearly in the elastic region, proceeded to the maximal point, and then decreased gradually. Finally a clear peak appeared in the buckling and shearing fracture. There is a curved relation between the specific gravity and the piezoelectric parameter when the influence of voids is considered, and there is a linear relation between the dynamic Young's modulus and the piezoelectric parameter when the stress is considered.Part of this paper was presented at the 46th annual meeting of the Japan Wood Research Society in Kumamoto, April 1996     

Effects of the lateral growth rate on wood quality parameters of Eucalyptus grandis from different latitudes in Brazil and Argentina

Climate change resulting from increased atmospheric carbon dioxide (CO₂) and shortages of fossil fuels such as petroleum are major problems worldwide. Under these conditions, demand for woody biomass resources is increasing. We investigated the feasibility of using fast-growing Eucalyptus grandis for material production. Samples of E. grandis were collected from four plantations in different latitude divisions, including tropical and subtropical Brazil and subtropical Argentina. Various xylem qualities were measured and related to the lateral growth rate. Lateral growth rate did not significantly affect the longitudinal released strain of the surface growth stresses or the xylem density at any of the sampling sites. Higher lateral growth rate, higher values of xylem density, and lower absolute values of the released strain were observed in plantations closer to the equator. Higher growth rates in tropical climate promote longer fiber length. In subtropical plantations, smaller diameter trees will produce tension wood with smaller microfibril angles. Planting E. grandis closer to the equator thus produces higher quality wood than in plantations at lower latitudes.     

Characterization of monoclonal antibodies against Actinobacillus pleuropneumoniae serotype 5.

Two monoclonal antibodies against Actinobacillus pleuropneumoniae serotype 5, designated as 5MAb-1 and 5MAb-6, were characterized. Enzyme-linked immunosorbent assay-inhibition tests with whole-cell antigens obtained from strains of serotype 1 through 12 of A pleuropneumoniae revealed that 5 MAb-1 bound to only serotype-5 strains. The epitope recognized by 5MAb-1 was a carbohydrate that was sensitive to periodate oxidation and resided on the structure of beta-1,6-linked D-galactose in an O-antigen polysaccharide of serotype-5 lipopolysaccharide. Analysis of these results revealed that the O-antigen polysaccharide of lipopolysaccharide was 1 of the antigenic determinants responsible for the serotype specificity of A pleuropneumoniae. On the other hand, 5MAb-6 reacted with strains of serotype 1 through 10 in varying degrees and its epitope was located on polypeptides sensitive to proteinase K. In an immunoblotting analysis, 5MAb-6 reacted with 2 polypeptide bands, with molecular weights of approximately 41,500 and 28,000, in the outer membrane protein-rich fraction obtained from strains of serotype 1 through 10. These results indicated that outer membrane proteins from several serotype strains of A pleuropneumoniae possessed common antigenic determinants.