Isolation and structural elucidation of some glycosides from the rhizomes of smaller galanga (Alpinia officinarum Hance)

Glycosidically bound compounds were isolated from the methanol extract of fresh rhizomes of smaller galanga (Alpinia officinarum Hance). Nine glycosides (1-9) were finally obtained by reversed-phase HPLC and their structures were elucidated by MS and NMR analyses. They were the three known glycosides, (1R,3S,4S)-trans-3-hydroxy-1,8-cineole beta-D-glucopyranoside (1), benzyl beta-D-glucopyranoside (3), and 1-O-beta-D-glucopyranosyl-4-allylbenzene (chavicol beta-D-glucopyranoside, 4); and the six novel glycosides, 3-methyl-but-2-en-1-yl beta-D-glucopyranoside (2), 1-hydroxy-2-O-beta-D-glucopyranosyl-4-allylbenzene (5), 1-O-beta-D-glucopyranosyl-2-hydroxy-4-allylbenzene (demethyleugenol beta-D-glucopyranoside, 6), 1-O-(6-O-alpha-L-rhamnopyranosyl-beta-D-glucopyranosyl)-2-hydroxy-4-allylbenzene (demethyleugenol beta-rutinoside, 7), 1-O-(6-O-alpha-L-rhamnopyranosyl-beta-D-glucopyranosyl)-4-allylbenzene (chavicol beta-rutinoside, 8), and 1,2-di-O-beta-D-glucopyranosyl-4-allylbenzene (9). Compounds 2-9 were detected for the first time as constituents of galanga rhizomes.     

Stimulatory effect of plasma samples from fattening cattle on adipogenesis‐related gene expression in preadipocyte cells

It is desirable to produce beef with high levels of monounsaturated fatty acids (MUFA), as this is related to fat softness and palatability. However, the physiology of MUFA synthesis in bovine fat during the fattening process remains to be established. In this study, in order to elucidate the relationship between plasma components and the fatty acid composition of intramuscular fat, we investigated the effect of plasma obtained from fattening cattle on the messenger RNA (mRNA) expressions of the adipogenesis‐related gene in a clonal bovine intramuscular preadipocyte line (BIP cells). The mRNA expressions of stearoyl‐CoA desaturase, adipocyte Protein 2, peroxisome proliferator‐activated receptor gamma and sterol regulatory element‐binding protein 1 in BIP cells were significantly higher following treatment with those plasma samples collected from the cattle with the highest diaphragmatic unsaturated fatty acids to saturated fatty acids ratio (US/S). Furthermore, the concentration of nonesterified fatty acid (NEFA) in the plasma samples had an inverse correlation with carcass diaphragmatic US/S. These results indicate that cattle with a low ratio of US/S in fat may be discriminated from the population of fattening cattle before slaughter by measuring the effect of their plasma on gene expression in BIP cells as well as their plasma concentration and composition of NEFA.     

Genetic alterations of KIT during clonal expansion and subsequent acquisition of resistance to toceranib in a canine mast cell tumor cell line

One of the potential mechanisms underlying acquired resistance to toceranib in canine mast cell tumor (MCT) is the emergence of a secondary mutation in the KIT gene. Here, genetic alterations of KIT during clonal expansion and subsequent acquisition of resistance to toceranib were investigated in the toceranib‐susceptible canine MCT cell line VI‐MC, which carries a KIT‐activating mutation resulting in a predicted p.(Asn508Ile) amino acid change in the receptor tyrosine kinase protein KIT. Two sublines were cloned from VI‐MC and toceranib‐resistant sublines then were established by continuous exposure to toceranib. The mutation status of KIT in parental VI‐MC and its sublines was investigated using next‐generation sequencing (NGS). Additionally, effects of secondary mutations on toceranib sensitivity in p.(Asn508Ile)‐mutant KIT were examined. KIT secondary mutations, including those encoding p.(Asn679Lys)‐, p.(Asp819Val)‐, and p.(Asp819Gly)‐mutant KIT, that confer toceranib insensitivity to p.(Asn508Ile)‐mutant KIT emerged only in toceranib‐resistant VI‐MCs. These mutations were not detected by NGS in the parental VI‐MC line or in the toceranib‐naive cloned VI‐MCs, although the parental line and sublines exhibited genetic heterogeneity in KIT that may have been caused by genetic evolution during clonal expansion. VI‐MC clones with these secondary mutations in KIT appear to have arisen from subclones during treatment with toceranib rather than being pre‐existing. However, further study using a higher resolution technique will be needed to confirm the developmental mechanism of KIT secondary mutation in canine MCT cells with acquired resistance to toceranib.     

Manufacture and properties of low-density binderless particleboard from kenaf core

Low-density binderless particleboards from kenaf core were successfully developed using steam injection pressing. The target board density ranged from 0.10 to 0.30g/cm³, the steam pressure used was 1.0MPa, and the steam treatment times were 7 and 10min. The mechanical properties, dimensional stability, and thermal and sound insulation performances of the boards were investigated. The results showed that the low-density kenaf binderless particleboards had good mechanical properties and dimensional stability relative to their low board densities. The board of 0.20g/cm³ density with a 10-min treatment time produced the following values: modulus of rupture 1.1MPa, modulus of elasticity 0.3GPa, internal bond strength 0.10MPa, thickness swelling in 24h water immersion 6.6%, and water absorption 355%. The thermal conductivity of the low-density kenaf binderless particleboards showed values similar to those of insulation material (i.e., rock wool), and the sound absorption coefficient was high. In addition, the boards are free from formaldehyde emission. Kenaf core appears to be a potential raw material for low-density binderless panels suitable for sound absorption and thermally resistant interior products.Part of this report was presented at the 52th Annual Meeting of the Japan Wood Research Society, Gifu, Japan, April 2002     

Establishment and characterization of primary canine hepatocellular carcinoma cell lines producing alpha-fetoprotein

A primary cultured cell line named CHKS was established from a hepatocellular carcinoma (HCC) of a dog showing a high level of serum alpha-fetoprotein (AFP). CHKS secreted a 66 KDD AFP into the growth medium regardless of the presence or absence of fetal bovine serum (FBS). Cloning CHKS with limiting dilution produced 4 clones, CHKS-1, -2, -3, and -4, which secreted 826, 471, 70, and less than 10 ng/ml, respectively, of AFP into the culture medium. In culture, these cell lines were similar in morphology and proliferation pattern to epithelial cells and positive to periodic acid-Schiff (PAS) staining. The presence of mRNA for canine albumin was demonstrated by nested PCR. The doubling times of the clone cell lines were 21, 45, 36, and 35 h, saturation densities 34, 18, 22, and 24 x 10(4)/cm(2), and plating efficiencies 18, 45, 46, and 45%, respectively. Chromosome analysis of these cell lines showed near triploidy. These results show that CHKS and its clones have hepatic cell functions and are useful for carcinogenetic and clinical studies of canine HCC.     

Pathogenic diversity of Phytophthora sojae and breeding strategies to develop Phytophthora-resistant soybeans

Phytophthora stem and root rot, caused by Phytophthora sojae, is one of the most destructive diseases of soybean [Glycine max (L.) Merr.], and the incidence of this disease has been increasing in several soybean-producing areas around the world. This presents serious limitations for soybean production, with yield losses from 4 to 100%. The most effective method to reduce damage would be to grow Phytophthora-resistant soybean cultivars, and two types of host resistance have been described. Race-specific resistance conditioned by single dominant Rps (“resistance to Phytophthora sojae”) genes and quantitatively inherited partial resistance conferred by multiple genes could both provide protection from the pathogen. Molecular markers linked to Rps genes or quantitative trait loci (QTLs) underlying partial resistance have been identified on several molecular linkage groups corresponding to chromosomes. These markers can be used to screen for Phytophthora-resistant plants rapidly and efficiently, and to combine multiple resistance genes in the same background. This paper reviews what is currently known about pathogenic races of P. sojae in the USA and Japan, selection of sources of Rps genes or minor genes providing partial resistance, and the current state and future scope of breeding Phytophthora-resistant soybean cultivars.     

The National BioResource Project (NBRP) Lotus and Glycine in Japan

The objective of the National BioResource Project (NBRP) in Japan is to collect, conserve and distribute biological materials for life sciences research. The project consists of twenty-eight bioresources, including animal, plant, microorganism and DNA resources. NBRP Lotus and Glycine aims to support the development of legume research through the collection, conservation, and distribution of these bioresources. Lotus japonicus is a perennial legume that grows naturally throughout Japan and is widely used as a model plant for legumes because of such advantages as its small genome size and short life cycle. Soybean (Glycine max) has been cultivated as an important crop since ancient times, and numerous research programs have generated a large amount of basic research information and valuable bioresources for this crop. We have also developed a “LegumeBase” a specialized database for the genera Lotus and Glycine, and are maintaining this database as a part of the NBRP. In this paper we will provide an overview of the resources available from the NBRP Lotus and Glycine database site, called “LegumeBase”.     

Therapeutic potential of canine bone marrow stromal cells (BMSCs) in the carbon tetrachloride (CCl4) induced chronic liver dysfunction mouse model

Regenerative medicine using bone marrow cells is an attractive therapy for the cure of patients with severe liver disease. Here, we show the therapeutic potential of canine bone marrow stromal cells (BMSCs) in mouse models of CCl(4)-induced chronic liver dysfunction. We used two different models for xenotransplantation, nude mice and cyclosporine A (CSA) immunosuppressed mice. Serum parameters from a standard liver panel were not improved following transplantation. However, fibrotic liver lesions with severe inflammation were decreased in CCl(4)-treated CSA mice following BMSC transplantation. Effective migration of transplanted canine BMSCs was limited to persistently injured liver in CCl(4)-treated CSA mice, where they may be effective in resolving inflammatory fibrotic lesions. These results suggest that canine BMSCs are an effective cell source for liver regeneration.     

Effect of steam treatment for the enzymic saccharification of waste mushroom medium after cultivation of shiitake mushroom (Lentinula edodes) and enokitake mushroom (Flammulina velutipes)

Waste mushroom medium of shiitake and enokitake (WS and WE, respectively) was steamed (StmWS and StmWE, respectively) and saccharized with cellulase to obtain glucose which can be feedstock of bioethanol or other bioproducts. Mushroom medium before cultivation of shiitake and enokitake (MS and ME, respectively) was also steamed (StmMS and StmME, respectively) and saccharized to compare with WS and WE. WS was stored at 15 °C for 1 month (SS), and SS was steamed (StmSS) and saccharized. The amount of glucose from StmSS was more than that from StmWS. The cellulose contained in StmSS and StmWE was saccharified earlier than that in StmMS and StmME, respectively. When StmSS and StmWE-added Meicelase of 5 and 3 FPU/g substrate (9.7 and 5.8 mg Meicelase/g substrate), respectively, was saccharized for 120 h, the saccharification ratio (the amount of glucose saccharized from cellulose/theoretical amount of glucose from cellulose) of them exceeded 80 %. The amount of glucose obtained from StmSS and StmWE was 311.0 and 207.7 mg/g substrate under the condition. It was found that the cellulose contained in mushroom medium was saccharized with less amount of cellulase by the influence of cultivation of shiitake or enokitake, storage of shiitake, and steaming.     

Plant regeneration by somatic embryogenesis from mature seeds of <Emphasis Type="Italic">Magnolia obovata</Emphasis>

We attempted to develop a method for the regeneration of plantlets from mature seeds of medically important Magnolia obovata via the induction of somatic embryogenesis in vitro. We initially cultured halves of mature seeds on either Murashige and Skoog (MS) medium or B₅ medium that contained 0, 1, 5 or 10 μM gibberellic acid (GA₃) for 1 month and then transferred the half-seeds to half-strength MS basal medium or B₅ basal medium for further culture in the absence of GA₃. Proembryogenic masses (PEMs) were observed 1 month after the transfer of the halved mature seeds to the medium without GA₃. The frequency of formation of PEMs was higher (28%) after initial culture in MS basal medium plus 1 μM GA₃ than in other tested media (0 or 4%). Somatic embryos that had been developed from PEMs were cultured on half-strength MS basal medium or B₅ basal medium for completion of maturation and then transferred to fresh aliquots of the same medium for initiation of germination. The frequency of germination, with the formation of normal primary leaves and roots, was above 80%. We transferred the somatic embryo-derived plantlets to soil for acclimatization and the plantlets continued to thrive.