Comparative studies of the quantification of genetically modified organisms in foods processed from maize and soy using trial producing

Seven types of processed foods, namely, cornstarch, cornmeal, corn puffs, corn chips, tofu, soy milk, and boiled beans, were trial produced from 1 and 5% (w/w) genetically modified (GM) mixed raw materials. In this report, insect resistant maize (MON810) and herbicide tolerant soy (Roundup Ready soy, 40-3-2) were used as representatives of GM maize and soy, respectively. Deoxyribonucleic acid (DNA) was extracted from the raw materials and the trial-produced processed food using two types of methods, i.e., the silica membrane method and the anion exchange method. The GM% values of these samples were quantified, and the significant differences between the raw materials and the trial-produced processed foods were statistically confirmed. There were some significant differences in the comparisons of all processed foods. However, our quantitative methods could be applied as a screening assay to tofu and soy milk because the differences in GM% between the trial-produced processed foods and their raw materials were lower than 13 and 23%, respectively. In addition, when quantitating with two primer pairs (SSIIb 3, 114 bp; SSIIb 4, 83 bp for maize and Le1n02, 118 bp; Le1n03, 89 bp for soy), which were targeted within the same taxon specific DNA sequence with different amplicon sizes, the ratios of the copy numbers of the two primer pairs (SSIIb 3/4 and Le1n02/03) decreased with time in a heat-treated processing model using an autoclave. In this report, we suggest that the degradation level of DNA in processed foods could be estimated from these ratios, and the probability of GM quantification could be experimentally predicted from the results of the trial producing.     

Uptake and translocation of bleaching herbicidal compounds in radish seedlings

The relationship between the hydrophobicity of certain herbicidal compounds and the bleaching pattern caused on radish cotyledons was investigated. Seed treatment with diphenylpyridones, as well as with established herbicides, produced three types of bleaching pattern according to their hydrophobicity. The less hydrophobic compounds caused complete bleaching of both cotyledons, but the compounds with more hydrophobicity caused only partial bleaching. The critical points for whole or partial bleaching were in the range of log K_ow 4–5 (K_ow: octanol/water partition coefficient), and these values were changed slightly with their chemical classes. Uptake of compounds into the seed coat took place rapidly; these compounds were then translocated slowly from the seed coat to the embryo, namely, radicle, abaxial surface of one cotyledon and marginal areas of both cotyledons. Application of these compounds to roots resulted in initial translocation to marginal areas of both cotyledons, with subsequent translocation to the middle area. It is believed that compounds taken up into the radicle were translocated to both cotyledons in a manner similar to that following application to roots. These effects following uptake by seeds can be used as a translaminal and lateral transport assay for bleaching herbicidal compounds in cotyledons.     

HPLC quantification of kaempferol-3-O-gentiobioside in Cassia alata

Kaempferol-3-O-gentiobioside, the major flavonoid glycoside in Indonesian Cassia alata was quantified in various parts of the plant. The mature leaf was found to contain the highest content of this metabolite. A decrease of the flavonoid content in the juvenile leaf during the period of October through December was also observed. The contents ranged from 2.0 to 5.0% and 1.0 to 4.0% in mature and juvenile leaves, respectively. The other parts studied were flower (sepal and petal), rachis, stem and seed. Kaempferol-3-O-gentiobioside was not detected in the seed.     

Seasonal changes in the xanthophyll cycle and antioxidants in sun-exposed and shaded parts of the crown of Cryptomeria japonica in relation to rhodoxanthin accumulation during cold acclimation

Xanthophyll rhodoxanthin, which is present in sun-exposed needles of certain gymnosperms in winter, may have a photoprotective role during long-term cold acclimation. To examine how cold acclimation processes vary within tree crowns and to examine putative correlations between xanthophyll cycle pigments (VAZ), rhodoxanthin and the water-water cycle in photoprotection, we monitored seasonal changes in the activities of two key antioxidant enzymes (ascorbate peroxidase (APX) and glutathione reductase (GR)), pigment composition and chlorophyll fluorescence parameters in sun and shade needles of crowns of the gymnosperm Cryptomeria japonica D. Don. Although APX and GR activities in both sun and shade needles were higher in winter than in summer when assayed at 20 degrees C, differences between seasons were less pronounced when enzymatic activities in summer and winter were assayed at 20 and 5 degrees C, respectively. These results suggest that increases in the potential activity of antioxidant enzymes in winter is an adaptation that helps counterbalance reductions in absolute enzyme activity caused by low temperature, and thus allows the photoprotective capacity of the water-water cycle in C. japonica to be maintained at a roughly constant value throughout the year. In shade needles, the concentration of VAZ increased in winter, but no rhodoxanthin accumulated. Photosynthetic activity was maintained in winter. In sun needles, however, the electron transport rate (ETR) and photochemical quenching (q(P)) decreased to their lowest values in December, just before the accumulation of rhodoxanthin, which coincided with the highest amount of VAZ. Changes in rhodoxanthin concentration mirrored changes in VAZ concentration from January to March. Winter values of ETR and q(P) were comparable with summer values after accumulation of rhodoxanthin, indicating that rhodoxanthin may play a more important role than the VAZ cycle in protecting the photosynthetic apparatus from photodamage in winter. Photosynthetic activity may be modulated, as a result of the interception of light by rhodoxanthin, to match the extent to which absorbed light energy can be utilized in winter when the VAZ cycle is unable to operate effectively because of low temperatures.     

Glycophorin A-knockout mice, which lost sialoglycoproteins from the red blood cell membrane, are resistant to lethal infection of Babesia rodhaini

Recent in vitro-based studies using several Babesia spp. have suggested that sialic acids and/or sialoglycoproteins on host red blood cells (RBCs) play an important role in their invasion of RBCs. In the present study, we analyzed the RBC characteristics of glycophorin A (GPA)-knockout mice and studied their in vivo susceptibility to lethal infection of Babesia rodhaini for the first time. In immunoblot and lectin blot analyses, glycoproteins containing O-linked oligosaccharides terminated with alpha2-3-linked sialic acids disappeared from the RBCs of GPA homozygous ((-/-)) mice. Flow cytometric analysis showed a remarkable reduction of Maackia amurensis lectin II binding to the surface of GPA(-/-) RBCs relative to control RBCs, indicating an appreciable loss of alpha2-3-linked sialic acids on the RBC surface of GPA(-/-) mice. Importantly, while B. rodhaini caused lethal infection in wild-type mice, the infected GPA(-/-) mice showed inhibition of parasite growth and eventually survived. These results indicate that RBC sialoglycoproteins lost in GPA(-/-) mice are involved in the in vivo growth of B. rodhaini, probably functioning as essential molecule(s) for the parasite invasion of host RBCs in the blood circulation.     

Radial variations of wood properties in <Emphasis Type="Italic">Casuarina equisetifolia</Emphasis> growing in Bangladesh

Radial variations of wood properties (basic density, fiber length, vessel element length, and compression strength) in plantation-grown Casuarina equisetifolia in Bangladesh were investigated for effective utilization of the wood. Samples disks at breast height were randomly collected from trees in a 10-year-old plantation in Cox’s Bazar Forest Division, Bangladesh. The basic density showed a near-constant value up to 30 mm from the pith and then rapidly increased up to 60 mm from the pith. The fiber length and vessel element length gradually increased from the pith to bark. When radial variation of wood properties was determined according to relative distance from the pith, similar radial patterns were observed among the sample trees, indicating that the wood properties in C. equisetifolia may be related to the growth rate. The compression strength parallel to the grain (CS) increased from the pith to bark. A significant positive correlation was found between the air-dried density and the CS. The results obtained indicated that wood around the pith has a relatively low density, and wood outside the pith area has a relatively high density, suggesting that it could be used as structural lumber. Part of this report was presented at the 58th Annual Meeting of Japan Wood Research Society, Tsukuba, March 2008     

Growth characteristics, stress-wave velocity, and Pilodyn penetration of 15 clones of 12-year-old Tectona grandis trees planted at two different sites in Indonesia

Tree improvement programs for teak (Tectona grandis) have mainly focused on breeding of trees with superior growth characteristics. However, improvement in wood quality should be included in breeding programs for high yield and high quality timber. In the present study, growth characteristics [stem diameter (D), tree height (H), and bole volume (V)], stress-wave velocity (SWV), and Pilodyn penetration (Py) were measured for 15 clones of 12-year-old teak trees planted at two different sites in Indonesia to clarify the variations in tree growth characteristics, SWV, and Py among clones, their repeatability, interaction between genotype and environment, and correlations between measured characteristics. Significant differences of all measured characteristics were found among 15 clones at both sites. Their repeatability showed relatively moderate to high values in both sites. These results indicate that these characteristics are closely related to genetic factors. Significant interaction between genotype and environment was found in all measured characteristics. In addition, SWV and Py showed lower interaction between genotype and environment than growth characteristics. No significant correlation was found between growth characteristics and SWV. These results suggest that wood properties and growth characteristics of teak trees can be improved by application of an appropriate tree breeding program.     

Sporiolides A and B,New Cytotoxic Twelve-Membered Macrolides from a Marine-Derived Fungus Cladosporium Species

Two new cytotoxic twelve-membered macrolides, sporiolides A (1) and B (2), were isolated from the cultured broth of a fungus Cladosporium sp., which was separated from an Okinawan marine brown alga Actinotrichia fragilis, and the structures were elucidated by spectroscopic data. Sporiolides A (1) and B (2) exhibited cytotoxicity against murine lymphoma L1210 cells. Spoliolide A (1) showed antifungal activity against Cryptococcus neoformans and Neurospora crassa.