Purification and characterization of a corrinoid compound from Chlorella tablets as an algal health food

Vitamin B(12) content of an algal health food, Chlorella tablets (Chlorella sp.), was determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor-chemiluminescence methods. The values of 200.9-211.6 microg/100 g dry weight determined by the chemiluminescence method were similar to the values (201.3-285.7 microg/100 g dry weight) determined by the microbiological method. A corrinoid compound was purified to homogeneity from the Chlorella tablets and characterized. The purified corrinoid compound was identified as vitamin B12, on the basis of silica gel 60 TLC, C18 reversed-phase HPLC, 1H NMR spectroscopy, and UV-Vis spectroscopy.     

A comparison of regression methods for estimating soil-atmosphere diffusion gas fluxes by a closed-chamber technique

Continuous changes in methane (CH₄) and carbon dioxide (CO₂) concentrations inside a closed chamber were measured on the forest floor at three sites: a deciduous forest and a coniferous forest in Hokkaido, Japan, and a birch forest in West Siberia, Russian Federation. Flux estimations by three types of regression methods, exponential, nonlinear, and linear, were examined using field-collected concentration data. The pattern of change with time of the gas concentration in the headspace differed, mainly according to site but also, to a lesser extent, according to the gas. This was a function of both the chamber height and surface soil property relating to soil gas diffusion and the gas concentration profile. Flux estimations did not differ statistically between the exponential and nonlinear methods for either gas at any site, because both of those regression methods were based on diffusion theory. However, the flux values estimated by linear regression were significantly different from those estimated by the other two methods for both CH₄ and CO₂ at the deciduous forest site and for CO₂ at the coniferous forest site. Shortening the chamber deployment period improved the linearity of the curve, but did not completely eliminate the error. Our results suggest that linear regression is not a good model of the change in headspace concentration with time.     

Sequential Leaching of Volcanic Soil to Determine Plant-Available Cations and the Provence of Soil Minerals Using Sr Isotopes

Sequential leaching experiments using H₂O, H₂O₂, NH₄Cl, and HCl were performed on surface soils on an andesite substrate at four sites in the Kawakami mountainous area, central Japan. The solutions extracted from the dehydrated soil by H₂O, H₂O₂, and NH₄Cl have relatively constant ratios with respect to Ca, Mg, and Sr, while they have variable ⁸⁷Sr/⁸⁶Sr ratios depending on the site. The elemental ratios and Sr isotopes in the extracted solution are different from those of the soil minerals but identical to those of the soil solution and the plants. Sr isotopic data indicate that the residues after extraction from fine-grained minerals by NH₄Cl and HCl are a mixture of acid-resistant minerals derived from bedrock and from arid areas in China. We suggest that there is a vital exchange of Sr and other cations between plants and the soil pool of exchangeable cations through the soil solution, while soil minerals, except chlorite, do not participate in the exchange reaction.     

A crossover in the mechanical response of nanocrystalline ceramics

Multimillion-atom molecular dynamics simulation of indentation of nanocrystalline silicon carbide reveals unusual deformation mechanisms in brittle nanophase materials, resulting from the coexistence of brittle grains and soft amorphous grain boundary phases. Simulations predict a crossover from intergranular continuous deformation to intragrain discrete deformation at a critical indentation depth. The crossover arises from the interplay between cooperative grain sliding, grain rotations, and intergranular dislocation formation similar to stick-slip behavior. The crossover is also manifested in switching from deformation dominated by indentation-induced crystallization to deformation dominated by disordering, leading to amorphization. This interplay between deformation mechanisms is critical for the design of ceramics with superior mechanical properties.     

Role for piRNAs and noncoding RNA in de novo DNA methylation of the imprinted mouse Rasgrf1 locus

Genomic imprinting causes parental origin-specific monoallelic gene expression through differential DNA methylation established in the parental germ line. However, the mechanisms underlying how specific sequences are selectively methylated are not fully understood. We have found that the components of the PIWI-interacting RNA (piRNA) pathway are required for de novo methylation of the differentially methylated region (DMR) of the imprinted mouse Rasgrf1 locus, but not other paternally imprinted loci. A retrotransposon sequence within a noncoding RNA spanning the DMR was targeted by piRNAs generated from a different locus. A direct repeat in the DMR, which is required for the methylation and imprinting of Rasgrf1, served as a promoter for this RNA. We propose a model in which piRNAs and a target RNA direct the sequence-specific methylation of Rasgrf1.     

General acid-base catalysis in the mechanism of a hepatitis delta virus ribozyme

Many protein enzymes use general acid-base catalysis as a way to increase reaction rates. The amino acid histidine is optimized for this function because it has a pK(a) (where K(a) is the acid dissociation constant) near physiological pH. The RNA enzyme (ribozyme) from hepatitis delta virus catalyzes self-cleavage of a phosphodiester bond. Reactivity-pH profiles in monovalent or divalent cations, as well as distance to the leaving-group oxygen, implicate cytosine 75 (C75) of the ribozyme as the general acid and ribozyme-bound hydrated metal hydroxide as the general base in the self-cleavage reaction. Moreover, C75 has a pK(a) perturbed to neutrality, making it "histidine-like." Anticooperative interaction is observed between protonated C75 and a metal ion, which serves to modulate the pK(a) of C75. General acid-base catalysis expands the catalytic repertoire of RNA and may provide improved rate acceleration.     

Growth characteristic and sink strength of fruit at different CO2 concentrations in a Japanese and a Dutch tomato cultivar

The aim of the present study was to elucidate how fruit growth was limited by the source and sink capacities in a Japanese (‘Momotaro York’) and a Dutch (‘Dundee’) tomato cultivar. The two cultivars were grown hydroponically with a high-wire system in greenhouses for 25 weeks, and the growth characteristics and sink strength of fruit were determined. Fruits were pruned to four (4F) or one (1F) per truss. The latter were used to determine potential fruit growth, an indicator of fruit sink strength. Growth was also determined under normal (LC) and enriched (HC, 700 μmol mol⁻¹) CO₂ concentrations to examine the effect of source enhancement on fruit production. In both cultivars under normal CO₂, the growth rate of fruit pruned to 4F per truss was lower than that in 1F, indicating that maximum potential fruit growth was not achieved. Under HC conditions, fruit growth rate of ‘Dundee’ achieved in 4F trusses was lower than that in 1F. In ‘Momotaro York’ in HC, fruit growth in 4F trusses was close to potential. This implies that fruit growth was source-limited irrespective of CO₂ concentrations in ‘Dundee’ cultivar while fruit growth in ‘Momotaro York’ under normal and enriched CO₂ conditions was limited by source and sink strengths, respectively. Adjustments of cultural practices including increasing fruit number per truss and/or genetic approaches to enhancing fruit sink strength by breeding may improve fruit yields of Japanese cultivars under high source/sink conditions.     

Characterization of corrinoid compounds from a Japanese black tea (Batabata-cha) fermented by bacteria

A Japanese fermented black tea (Batabata-cha) contained a considerable amount of vitamin B(12) (456 +/- 39 ng per 100 g dry tea leaves and 2.0 +/- 0.3 ng per 100 mL of tea drink). A corrinoid compound was partially purified and characterized from the tea leaves. The patterns of the purified compound by the silica gel 60 thin-layer chromatography and C18 reversed phased high-performance liquid chromatography were identical to those of authentic vitamin B(12). When 20 week old vitamin B(12) deficient rats, which excreted substantial amounts (about 250 mg/day) of methylmalonic acid in urine as an index of vitamin B(12) deficiency, were fed the tea drink (50 mL/day, 1 ng of vitamin B(12)) for 6 weeks, urinary methylmalonic acid excretion (169 +/- 29 mg/day) of the tea drink-supplemented 26 week old rats decreased significantly relative to that (250 +/- 32 mg/day) of the deficient rats. The results indicate that the vitamin B(12) found in the fermented black tea is bioavailable in mammals.     

Effect of the number of intercrosses on grain yield in basic rice synthetic populations

The effect of 0, 1, 2, 3 and 4 intercross cycles using genetic male sterility on the mean yield and genetic variability of the CNA 5 irrigated rice synthetic population was assessed studying 300 S_0:2 (1998/99) and S_0:3 (1999/00) families grown in two locations (Lambari, MG and Goianira, GO). Each intercross was investigated using a 8 × 8 triple lattice design experiment containing 60 families and four control cultivars. Five experiments were carried out per location and generation, totaling 20 experiments. Individual, combined over locations, combined over generations and combined over locations and generations analyses of variance were performed. Linear regression equations were fitted to data of each location and generation as well as, genetic and phenotypic parameters were estimated. An average increase of 3.91% in grain yield was obtained with the increase in the number of intercrosses. However, when genetic gains obtained from selection of the five and ten highest yielding families were considered, negative b estimates were always obtained, suggesting a tendency of gain increase reduction with the increase in the number of intercrosses. This was confirmed by the analysis of the means of the best families, since increases in the number of intercrosses did not result in selection of families with greater mean grain yield. The estimates of the genetic variances and heritabilities did not suggest an increase in genetic variability with the increase in intercross cycles. Therefore, carrying out intercrosses on the base population did not improve the results of the irrigated rice recurrent selection program. This revised version was published online in August 2006 with corrections to the Cover Date.     

Morphology and water and lipid contents of stifle menisci of growing swine.

Morphology and distribution of water and lipid were examined in menisci of growing swine. The broadness of menisci and the size of protruded tissue in the inner anterior portion of menisci varied among animals. Small tears of tissues were frequently observed. Both the thickness and tissue weight were greater in the lateral than in the medial meniscus. Water content and intensity of glycosaminoglycan staining with safranin-O were found to be greatest in the inner one-third of the meniscus, and greater in the middle than in the outer one-third of the meniscus. Lipid content was greater in the outer one-third than in the inner two-thirds of the meniscus.