Estimating the equilibrium position by measuring growth stress in weeping branches ofPrunus spachiana Kitamura f.spachiana cv.Plenarosea

The equilibrium position of a current-year branch of the weeping Japanese cherry,Prunus spachiana Kitamura f.spachiana cv.Plenarosea, was estimated by measuring the released strains of growth stresses. In current-year branches that were supported with wires to prevent weeping as soon as the branches first budded, tensile growth stresses on the upper side were smaller than those of the control branches. Gelatinous fibers were rarely found on the upper part of the cross section of the supported branches, whereas the control branches had many gelatinous fibers on the upper part. The upright orientation of the supported branches was closer to the equilibrium position than the weeping orientation of the control branches. The equilibrium position of the branches was thought to be in the initial bud direction, above the horizontal plane; and the weeping style of branch was not the preferred angular orientation forP. spachiana.     

Effect of Fucoidan Extracted from Mozuku on Experimental Cartilaginous Tissue Injury

We investigated the effect of fucoidan, a sulfated polysaccharide, on acceleration of healing of experimental cartilage injury in a rabbit model. An injured cartilage model was surgically created by introduction of three holes, one in the articular cartilage of the medial trochlea and two in the trochlear sulcus of the distal femur. Rabbits in three experimental groups (F groups) were orally administered fucoidan of seven different molecular weights (8, 50, 146, 239, 330, 400, or 1000 kD) for 3 weeks by screening. Control (C group) rabbits were provided water ad libitum. After the experimental period, macroscopic examination showed that the degree of filling in the fucoidan group was higher than that in the C group. Histologically, the holes were filled by collagen fiber and fibroblasts in the C group, and by chondroblasts and fibroblasts in the F groups. Image analysis of Alcian blue- and safranin O-stained F-group specimens showed increased production of glycosaminoglycans (GAGs) and proteoglycans (PGs), respectively. Some injured holes were well repaired both macroscopically and microscopically and were filled with cartilage tissues; cartilage matrices such as PGs and GAGs were produced in groups F 50, F 146, and F 239. Thus, fucoidan administration enhanced morphologically healing of cartilage injury.     

Homologue gene of bile acid transporters ntcp,asbt, and ost-alpha in rainbow trout Oncorhynchus mykiss: tissue expression,effect of fasting,and response to bile acid administration

Bile acid transporters belonging to the SLC10A protein family, Na+ taurocholate cotransporting polypeptide (NTCP or SLC10A1), apical sodium-dependent bile salt transporter (ASBT or SLC10A2), and organic solute transporter alpha (Ost-alpha) have been known to play critical roles in the enterohepatic circulation of bile acids in mammals. In this study, ntcp, asbt, and ost-alpha-1/-2 cDNA were cloned, their tissue distributions were characterized, and the effects of fasting and bile acid administration on their expression were examined in rainbow trout Oncorhynchus mykiss. The structural characteristics of Ntcp, Asbt, and Ost-alpha were well conserved in trout, and three-dimensional structure analysis showed that Ntcp and Asbt were similar to each other. Tissue distribution analysis revealed that trout asbt was primarily expressed in the hindgut, while ntcp expression occurred in the brain, and ost-alpha-1/-2 was mainly expressed in the liver or ovary. Although asbt and ost-alpha-1 mRNA levels in the gut increased in response to fasting for 4 days, ost-alpha-1 expression in the liver decreased. Similarly, bile acid administration increased asbt and ost-alpha-1 expression levels in the gut, while those of ntcp and ost-alpha-2 in the liver decreased. These results suggested that the genes asbt, ntcp, and ost-alpha are involved in bile acid transport in rainbow trout.     

Rapid genotyping with DNA micro-arrays for high-density linkage mapping and QTL mapping in common buckwheat (Fagopyrum esculentum Moench)

For genetic studies and genomics-assisted breeding, particularly of minor crops, a genotyping system that does not require a priori genomic information is preferable. Here, we demonstrated the potential of a novel array-based genotyping system for the rapid construction of high-density linkage map and quantitative trait loci (QTL) mapping. By using the system, we successfully constructed an accurate, high-density linkage map for common buckwheat (Fagopyrum esculentum Moench); the map was composed of 756 loci and included 8,884 markers. The number of linkage groups converged to eight, which is the basic number of chromosomes in common buckwheat. The sizes of the linkage groups of the P1 and P2 maps were 773.8 and 800.4 cM, respectively. The average interval between adjacent loci was 2.13 cM. The linkage map constructed here will be useful for the analysis of other common buckwheat populations. We also performed QTL mapping for main stem length and detected four QTL. It took 37 days to process 178 samples from DNA extraction to genotyping, indicating the system enables genotyping of genome-wide markers for a few hundred buckwheat plants before the plants mature. The novel system will be useful for genomics-assisted breeding in minor crops without a priori genomic information.     

Histological analysis of implanted embryos in large Japanese field mouse (Apodemus speciosus) and estimation of developmental stage

The large Japanese field mouse (Apodemus speciosus) is a small rodent species endemic to Japan. The genetic characteristics of A. speciosus include different chromosome numbers within the same species. Furthermore, A. speciosus has been used in radiation and genetic research. In the present study, a pregnant A. speciosus was obtained, and histochemical analysis of the implanted embryos was performed and compared with the developmental stages of the mouse (Mus musculus). Although there were some differences, the structures of the implanted embryos, including the primitive streak and placenta of A. speciosus were similar to those of mouse. Our study will be important for the construction of a developmental atlas of A. speciosus.     

Evaluation of table grape genetic resources for sugar,organic acid,and amino acid composition of berries

Variations in the sugar, organic acid, and amino acid composition of table grape berries were determined in three successive years in 129 grape cultivars. Three regional groups were represented among these cultivars; European, North American, and Japanese. All cultivars are conserved at the Fukuoka Agricultural Research Center in Japan. Three representative ripe average-sized bunches were selected from one vine per cultivar. Ripeness was judged by visual observation of seed color change to dark brown, without senescence of berry tissue. Soluble solids content (SSC) was weakly correlated with other berry traits, but strongly correlated with total sugar content. Genotypic variance (σ _g ²) was significant for all berry traits, especially for α ratio (sugar composition) and γ ratio (amino acid composition). The yearly effect (σ _y ²) was significant for SSC, total sugar content, and total amino acid content but not for α ratio, titratable acidity, β ratio (organic acid composition) and γ ratio. Repeatabilities over the 3 years were very high for α ratio and γ ratio, while they were low for SSC, total sugar content, titratable acidity, β ratio, and amino acid content. Correlation coefficients between the values of each individual year and the 3-year mean were relatively high for the α ratio and γ ratio. We highlight the varietal differences in the α ratio and γ ratio because of the low environmental variation. Two types of grape were distinguished on the basis of sugar composition: hexose accumulators (α ratio ≥ 0.81) and sucrose accumulators (α ratio ≤ 0.80). The hexose- and sucrose-accumulating traits were related to genetic or regional differences. Cultivars were classified into five types on the basis of the γ ratio. Like the α ratio, variation in the γ ratio is closely related to cultivar parentage and regional differences. Vitis vinifera cultivars exhibited a skewed distribution with a γ ratio below 1.0 because they contain low levels of sweet amino acids. In contrast, the γ ratio frequently exceeded 1.5 in American hybrid cultivars because they contain high levels of sweet amino acids.     

Myeloma-related disorder with leukaemic progression in a cat

A 10-year-old American Shorthair cat with nasal discharge, anorexia, and weight loss was found to have pancytopenia and hyperproteinaemia. Bone marrow aspiration revealed atypical plasma cells that totalled 50% of the nucleated bone marrow cells. The number of atypical plasma cells progressively increased in the peripheral blood during the observation period of 64 days. The cat did not respond to treatments with melphalan, chlorambucil, and prednisolone, and died 71 days after the initial presentation. Clinical, cytological, histopathological, and immunohistochemical findings in this case supported the diagnosis of myeloma-related disorder (MRD) with leukaemic progression.     

New Cyclic Lipopeptides of the Iturin Class Produced by Saltern-Derived Bacillus sp. KCB14S006

Salterns, one of the most extreme natural hypersaline environments, are a rich source of halophilic and halotolerant microorganisms, but they remain largely underexplored ecological niches in the discovery of bioactive secondary metabolites. In continued efforts to investigate the metabolic potential of microbial populations from chemically underexplored sites, three new lipopeptides named iturin F₁, iturin F₂ and iturin A₉ (1–3), along with iturin A₈ (4), were isolated from Bacillus sp. KCB14S006 derived from a saltern. The structures of the isolated compounds were established by 1D-, 2D-NMR and HR-ESIMS, and their absolute configurations were determined by applying advanced Marfey’s method and CD spectroscopy. All isolates exhibited significant antifungal activities against various pathogenic fungi and moderate cytotoxic activities toward HeLa and src^ts-NRK cell lines. Moreover, in an in vitro enzymatic assay, compound 4 showed a significant inhibitory activity against indoleamine 2,3-dioxygenase.     

Mortality of planted <Emphasis Type="Italic">Pinus thunbergii</Emphasis> Parlat. saplings subject to coldness during winter and soil types in region of seasonal soil frost

To assess and improve methods for the development of coastal forests in Hokkaido, northern Japan, this study examined the factors affecting mortality of planted Pinus thunbergii Parlat. saplings in a region with seasonal soil frost. The needles of pine saplings at the study site turn red in late spring, suggesting xylem embolism, which can lead to death. Sapling mortality was strongly correlated with the degree of cold just after planting and with the occurrence of repeated severe winters. Volcanic ash soil had been supplied to improve the soil nutrients, and the volcanic ash soil always thawed later than the original coastal sand. Saplings in volcanic ash were significantly more damaged than those in original coastal sand. This suggests that hydraulic stress caused by cuticular transpiration under frozen soil conditions during spring may also accelerate damage due to embolisms. Hence, volcanic ash soil should not be applied to soils in regions with seasonal soil frost. A covering of woodchips can help prevent frost from reaching greater soil depths, and, indeed, fewer saplings at the study site died in areas with such coverage. Planting location can also affect pine sapling mortality. For example, shading caused by wood fencing may affect the soil freezing profile during winter; therefore, planting close to a fence should also be avoided in regions with seasonal soil frost. These findings should be used to improve the development of forests in regions with seasonal soil frost.     

Enzyme-linked immunosorbent assay for total sennosides using anti-sennside A and anti-sennoside B monoclonal antibodies

Total sennosides concentration is a very important factor when rhubarb and senna will be used as crude drugs. However, one-step analytical technique for total sennosides has not been reported except HPLC. An enzyme-linked immunosorbent assay (ELISA) for total sennosides concentration by using the combination of anti-sennoside A (SA) and anti-sennoside B (SB) monoclonal antibodies (MAbs) in a single assay has been investigated. Total sennosides concentration in rhubarb and senna samples determined by newly developed assay system showed good agreement with those analyzed by ELISA using anti-SA MAb and anti-SB MAb, respectively.