Balenine,imidazole dipeptide,induces activation of superoxide dismutase in myotubes

Balenine is one of the endogenous imidazole dipeptides. It is composed of beta-alanine and 3-methyl-l-histidine, which exist mainly in the muscles and the brain. The exact biological properties of balenine are still not well known, although the antioxidant activity of carnosine, another imidazole dipeptide, is known. In this study we investigated whether balenine exhibits antioxidant activity. It was found to decrease the superoxide anion (O₂⁻) and increased hydrogen peroxide (H₂O₂) generation. We found that SOD activity increased in balenine-treated C2C12 myotubes, although balenine did not increase expression of SOD mRNA. On the other hand, there were no changes in other antioxidant enzymes, CAT and GPX activity, and mRNA levels. In an in vitro assay, the direct activation of SOD treated by balenine was significantly higher than with carnosine treatment. Moreover, balenine constituent amino acids did not have the ability to activate SOD. Our results suggest that balenine contributes to antioxidant effects through activation of SOD.

     

Purification of anaphase promoting complex/cyclosome from goldfish oocytes

Destruction of cyclin B is required for exit from mitosis and meiosis. A cyclin-specific ubiquitinating system, including anaphase-promoting complex/cyclosome (APC/C) is thought to be responsible for cyclin B destruction. To learn more about the molecular mechanism of cyclin B degradation, a molecular study of the ubiquitinating system in goldfish has been undertaken. For biochemical preparation of APC/C, we first conducted the cloning, sequencing and expression analysis of goldfish, Carassius auratus, cdc27 that encodes a subunit of APC/C from goldfish ovary. The deduced amino acid sequence is highly homologous to cdc27 from other species. Then recombinant goldfish Cdc27CT (C-terminal half of Cdc27) was expressed in Escherichia coli, and an antibody was raised against purified recombinant protein. Polyclonal antiserum cross-reactive with Cdc27 was obtained. By the assay using the antibody, APC/C was purified by column chromatographs.     

Nondestructive detection of cracks near the surface of wooden boards by dynamic heat dissipation

A method to detect cracks near the surface of wooden boards has been developed where the change in surface temperature of the wood sample is monitored using an infrared camera following momentary heating by a flash. Cylindrical holes simulating cracks were drilled into the wood samples, and blackbody paint was painted onto the surface of the samples to assist flash absorption. This method uses the dynamic heat transport process from the blackbody paint to the surface of the wood sample to the cracks over a short timescale. The theoretical foundations of the observation method were outlined, and the technique was verified in experiments with samples of Cercidiphyllum japonicum, Western red cedar, Japanese cedar, Balsa, and medium-density fiberboard. The developed technique is able to detect the presence of holes located near the surface of some samples. However, this method could not detect the presence of holes in Balsa nor holes located deeper than 1 mm from the surface of the sample. A theoretical analysis of these phenomena was provided to help interpreting them.     

Cytoplasmic relatedness of apple landraces and cultivars: a molecular analysis

Summary Mitochondrial DNA (mtDNA) diversity has been determined in apple landraces and cultivars, based upon restriction fragment length polymorphisms (RFLPs) for several mitochondrial genes. Our analysis includes three cultivars, Golden Delicious, McIntosh and Delicious, which represent the various patterns of mtDNA polymorphisms previously described (Ishikawa et al., 1992). A total of five RFLPs were detected, allowing classification of the apple genotypes into four cytoplasmic groups: GroupI, Golden Delicious-type; GroupII, McIntosh-type; GroupIII, Delicious-type; and Group IV, Dolgo Crab-type. European landraces and cultivars were assigned to Groups I, II, and III, while chinese crab apples were placed in either Group III or IV.     

Evaluation of genetic variation among wild rice populations in Cambodia

A total of 448 samples in five natural populations of wild rice (Oryza rufipogon) were collected in Cambodia. They were examined using 12 SSR and two chloroplast markers to evaluate the degree of variation among populations and the genetic structure within populations. In the two annual populations, the number of plants with homozygous alleles at all 12 SSR loci were high (66.3% and 79.5%), suggesting that these plants propagate mainly through self-pollination. In the three perennial populations, no individuals had all homozygous genotypes, but redundant genotypes resulted from clonal propagation were observed. Percentages of the redundant genotypes were highly varied (3.6%, 29.2% and 86.0%). This may be due to the different stable levels of environmental conditions. As for chloroplast genome, most of the wild plants showed the same chloroplast types as most Indica-type cultivars have. However, plants with different chloroplast types were maintained, even in the same population. In tropical Asian countries, many wild rice populations were observed under similar ecological conditions examined in this study. Therefore, the present results concerning population structure will be important to further elucidate genetic features of wild rice, and will also give strong clues to utilize and conserve wild natural genetic resources.     

Analysis of wheat-Thinopyrum intermedium derivatives with BYDV-resistance

Chromosome compositions of seven lines, derived from hybrids between a wheat cultivar and the wheat-Thinopyrum intermedium addition line Z6, with barley yellow dwarf virus (BYDV) resistance, were determined by genomic in situ hybridization, cytogenetic and SSR assays. The results showed that line N522 was a disomic addition line, lines N420 and N439 were 2Ai-2(2B) chromosome substitution lines, lines N431 and N452 were 2Ai-2(2D) chromosome substitution lines, line N523 was a 2Ai-2S(2D) ditelosomic substitution line, and line N530 was a double ditelosomic line with the mitotic chromosome number of 2n = 40 + 4t. One pair of telosomes in line N530 lacked several proximal SSR markers of chromosome 2AS, but possessed certain terminal markers, which were consistent with an acrocentric structure, and the other pair of chromosome arms were presumably 2Ai-2S telosomes with BYDV-resistance. These wheat-Th. intermedium lines provide useful genetic resources for developing alien chromosome translocation lines.     

Mitochondrial DNA Restriction Fragment Length Polymorphisms in Malus Species

Three apple (Malus×domestica) cultivars and 11 Malus accessions have been investigated by the probe hybridization method on their mitochondrial DNA (mtDNA). The gene probes used were: coxI, coxII, atpA, atp6, and atp9. Our results revealed enough variation to characterize ten mtDNA haplotypes among the Malus genotypes examined. The taxonomic and phylogenetic implications of mtDNA polymorphism are also discussed.     

Evaluation of biological activities of extracts from the fruiting body of <Emphasis Type="Italic">Pleurotus citrinopileatus</Emphasis> for skin cosmetics

Pleurotus citrinopileatus Singer has recently become a popular delicacy in East Asian countries. We prepared a methanol extract, soluble fractions from the methanol extract, and a hot water extract of the fruiting bodies of P. citrinopileatus. The biological activities such as melanin biosynthesis inhibition, tyrosinase inhibition, antioxidant activities [1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, oxygen radical absorbance capacity (ORAC), and superoxide dismutase (SOD)-like activity], antibacterial activities, and antihyaluronidase activities of these extracts were evaluated. We found that the n-hexanesoluble, diethyl ether-soluble, and ethyl acetate-soluble fractions exhibited melanin biosynthesis inhibition in B16 melanoma cells, as well as antioxidant (ORAC) and antibacterial activities. However, the n-butanol-soluble and water-soluble fractions and the methanol and hot water extracts exhibited antioxidant (DPPH radical scavenging, SOD-like activity) and antihyaluronidase activities. These results indicate that the fruiting bodies of P. citrinopileatus have the potential to be used as an ingredient in skin cosmetics.     

Identification of a novel cytotoxic protein, Cry45Aa, from Bacillus thuringiensis A1470 and its selective cytotoxic activity against various mammalian cell lines

Parasporal inclusion proteins produced by Bacillus thuringiensis strain A1470 exhibit strong cytotoxicity against human leukemic T cells when activated by protease treatment. One of the cytotoxic proteins was separated by anion exchange chromatography and gel filtration chromatography and designated Cry45Aa. Its gene was then expressed in recombinant Escherichia coli, in which the Cry45Aa precursor was accumulated in an inclusion body. It was solubilized in sodium carbonate buffer and processed with proteinase K, and cytotoxic activities of the protein against various mammalian cell lines were evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide assay. The protein exhibited high cytotoxic activity against CACO-2, Sawano, MOLT-4, TCS, and HL60 cells and moderate activity against U-937 DE-4, PC12, and HepG2 cells. On the other hand, the EC50 values against Jurkat, K562, HeLa, A549, Vero, COS-7, NIH3T3, CHO, and four normal tissue cells (human primary hepatocyte cells, UtSMC, MRC-5, and normal T cells) were >2 microg/mL.