In vitro and in vivo consequences of VLA-2 expression on rhabdomyosarcoma cells

Cloned integrin alpha 2 subunit complementary DNA was expressed on human rhabdomyosarcoma (RD) cells to give a functional VLA-2 (alpha 2 beta 1) adhesion receptor. The VLA-2-positive RDA2 cells not only showed increased adhesion to collagen and laminin in vitro, but also formed substantially more metastatic tumor colonies in nude mice after either intravenous or subcutaneous injection. These results show that a specific adhesion receptor (VLA-2) can markedly enhance both experimental and spontaneous metastasis. In contrast to the metastasis results, there was no difference in either the in vitro growth rate or apparent in vivo tumorigenicity of RD and RDA2 cells.     

Variations in cryptic assemblages in coral-rubble interstices at a reef slope in Ishigaki Island,Japan

The interstices of coral rubble provide a good refuge for cryptic small crustaceans and molluscs, which are important prey items for reef fishes. In this study, we focused on the assemblage composition of cryptic mobile species along the depth gradient at an outer reef slope. Traps with coral rubble (22 × 18 × 6 cm) were set at three sites at different depths, and mobile animals that colonized the traps were sorted. In total, 117 species of larger animals (mainly Decapoda and Gastropoda) were recorded: 71 species at the 3 m point, 53 species at the 11 m point, and 37 species at the 22 m point. On average, 260.3 individuals were captured in the traps at 3 m, while 95.3 and 57.8 individuals were captured at 11 and 22 m, respectively. MDS ordination showed that assemblage composition differed among the three sites. The most abundant species at the 3 m was the galathean crab Galathea mauritiana, followed by the gastropod Diala albugo. At 11 and 22 m, the galathean crab Sadayoshia edwardsii was most abundant. Thus, at the reef slope, the benthic assemblage was most diverse at the shallow site, and specific communities occurred at the three sites.     

Direct genotyping of single pollen grains of a self-compatible mutant of Japanese pear (Pyrus pyrifolia) revealed inheritance of a duplicated chromosomal segment containing a second S-haplotype

Radiation mutant 415-1, which is the first known diploid pollen-part self-compatible mutant of pears (Pyrus spp.), has a decreased ability to produce pollen. To determine whether the self-compatibility trait is associated with this defect, we directly analyzed the genotypes of individual pollen grains by using polymerase chain reaction amplification of DNA from single pollen grains. We isolated single pollen grains from 415-1 and succeeded in genotyping the S-RNase gene and three simple sequence repeat (SSR) markers in linkage group 17. Out of 173 individual pollen grains, 28 (16 %) were S-heteroallelic. These pollen grains had two alleles each of the S-RNase gene and of two linked SSR loci, all on a duplicated chromosomal segment, but only one allele of a non-duplicated locus farther away on the same chromosome. The segregation ratio of each marker in the pollen from 415-1 was approximately the same as that observed in outcross progeny. This suggests that the decrease in frequency of pollen with the duplicated S-haplotype occurred during meiosis or pollen formation, but that the probability of fertilization by S-heteroallelic pollen is equal to that of single-allelic pollen. However, the partial sterility in 415-1 can also be attributed to one or more unidentified lethal mutations unlinked to the duplicated segment encompassing the S-haplotype. Single-pollen genotyping can be used in a variety of applications in genetic research because in cases where all pollen genotypes are proportionately represented in the progeny, segregation ratios can be obtained without producing the next generation.     

Identification of QTLs controlling harvest time and fruit skin color in Japanese pear (Pyrus pyrifolia Nakai)

Using an F₁ population from a cross between Japanese pear (Pyrus pyrifolia Nakai) cultivars ‘Akiakari’ and ‘Taihaku’, we performed quantitative trait locus (QTL) analysis of seven fruit traits (harvest time, fruit skin color, flesh firmness, fruit weight, acid content, total soluble solids content, and preharvest fruit drop). The constructed simple sequence repeat-based genetic linkage map of ‘Akiakari’ consisted of 208 loci and spanned 799 cM; that of ‘Taihaku’ consisted of 275 loci and spanned 1039 cM. Out of significant QTLs, two QTLs for harvest time, one for fruit skin color, and one for flesh firmness were stably detected in two successive years. The QTLs for harvest time were located at the bottom of linkage group (LG) Tai3 (nearest marker: BGA35) and at the top of LG Tai15 (nearest markers: PPACS2 and MEST050), in good accordance with results of genome-wide association study. The PPACS2 gene, a member of the ACC synthase gene family, may control harvest time, preharvest fruit drop, and fruit storage potential. One major QTL associated with fruit skin color was identified at the top of LG 8. QTLs identified in this study would be useful for marker-assisted selection in Japanese pear breeding programs.     

Histopathological, immunohistochemical and ultrastructural studies of a renal mesenchymal tumor in a young beagle dog

A 15-month-old male beagle dog used in a toxicity study had a primary renal mesenchymal tumor. Macroscopically, the tumor was a gray-white mass which was found in the right kidney, and extended from the capsule to a position slightly compressing the medulla. Microscopically, most of the tumor cells showed a myxoid pattern, in which the matrix was positive for alcian blue staining. In the other parts of the tumor, a fascicular and wavy pattern was observed, and the matrix was full of collagen fibrils. Immunohistochemically, tumor cells were positive for vimentin and fibronectin, and negative for cytokeratin, desmin, α-smooth muscle actin, Von Willebrand factor, cyclooxigenase-2 and myelin basic protein. As a result, we diagnosed this case to be a renal mesenchymal tumor. Based on the microscopic findings, interstitial characteristics and immunohistochemical features, the present case was classified as a congenital mesoblastic tumor.     

Characterization of lignin-derived products from various lignocellulosics as treated by semi-flow hot-compressed water

To elucidate the decomposition behaviors of lignin from different taxonomic groups, five different lignocellulosics were treated with hot-compressed water (230 °C/10 MPa/15 min) to fractionate lignins into water-soluble portions, precipitates, and insoluble residues. The lignin-derived products in each fraction were characterized and compared. The delignification of monocotyledons [nipa palm (Nypa fruticans) frond, rice (Oryza sativa) straw, and corn (Zea mays) cob] was more extensive than that achieved for Japanese cedar (Cryptomeria japonica, gymnosperm) and Japanese beech (Fagus crenata, dicotyledon angiosperm). The water-soluble portions contained lignin monomers like coniferyl alcohol and phenolic acids, while the precipitates contained higher molecular weight lignin with high content of ether-type linkages. Lignin in the insoluble residues was rich in condensed-type structures. In all five lignocellulosics, ether-type linkages were preferentially cleaved, while condensed-type lignin showed resistance to hot-compressed water. In the monocotyledons, lignin–carbohydrate complexes were cleaved and gave lignins that had higher molecular weights than those eluted from the woods. These differences would facilitate the delignification in monocotyledons. Such information provides useful information for efficient utilization of various lignocellulosics.     

Neurotropism of the 1997 Hong Kong H5N1 influenza virus in mice

The direct transmission of H5N1 influenza A viruses from chickens to humans in Hong Kong in 1997 emphasized the need to have information on the pathogenesis of avian influenza virus infection in mammals. H5N1 influenza viruses isolated from patients during the incident killed experimentally infected mice. The principal lesions of the mice were broncho-interstitial pneumonia and nonsuppurative encephalitis. Infectious viruses and/or viral antigens were detected in the brain as well as in the trigeminal and vagal ganglia but not in the blood of the mice. These findings suggest that the virus reached the brain through the vagus and/or trigeminal nerves following replication in the respiratory mucosa. The results imply that neurotropism of the H5N1 virus in mice is a novel characteristic in the pathogenesis of infection by human influenza virus isolates.     

Protective effects of intranasal vaccination with plasmid encoding pseudorabies virus glycoprotein B in mice

Intranasal administration of plasmid DNA encoding glycoprotein B of pseudorabies virus into mice induced both serum and secretory antibody responses. These mice resisted intranasal challenge with lethal dose of the virus, but did not intraperitoneal challenge. On the other hand, intramuscular injection of the plasmid induced less secretory and higher serum antibody responses than those of intranasally vaccinated mice. None of them was protected from virus challenge. The present results suggest that administration of plasmid DNA encoding glycoprotein B by respiratory mucosal route generates local secretory antibodies which serve to protect animals from pseudorabies virus infection.     

Acid phosphatase in cheese

The combined action of proteinases/peptidases and acid phosphatases on phosphorylated proteins/peptides could be important for cheese quality. It seems that acid phosphatase activity in cheese originates from milk and microorganisms. Bovine milk contains more than one acid phosphatase, which are distributed between the cream and skim milk. The properties of two enzymes isolated from butter milk and skim milk are different. In general, acid phosphatase, especially that from starter organisms is very heat stable. The dephosphorylation of casein and phosphopeptides is of interest in cheese ripening. It has been suggested that acid phosphatases from both skim milk and lactic acid bacteria contribute to dephosphorylation of phosphopeptides in cheese, which are produced from casein by coagulant, indigenous milk proteinase and microbial proteinases during cheese ripening.