DNA methylation status of bovine blastocyst embryos obtained from various procedures

DNA methylation is an important factor for the regulation of gene expression in early embryos. It is well known that the satellite I sequence is more heavily methylated in bovine somatic cell nuclear transfer (NT-SC) embryos than in embryos derived from in vitro fertilization (IVF). However, the methylation status of bovine embryos obtained by other procedures is not well known. To clarify DNA methylation levels of bovine embryos obtained from various procedures, we examined satellite I sequences in bovine blastocyst (BC) embryos derived from NT-SC, NT using embryonic blastomeres (NT-EM), in vivo (Vivo), IVF and parthenogenetic treatment (PA). Furthermore, in order to evaluate the efficacy of DNA demethylation by the NT procedure, we determined the DNA methylation levels in bovine embryos in which NT was recapitulated (Re-NT). Although the DNA methylation levels in the NT-SC embryos were higher than those in the other embryos, the NT-EM embryos exhibited lower DNA methylation levels. The satellite I sequence in the NT-SC embryos was more demethylated than that in the donor cells. Although the DNA methylation level in the individual NT-SC embryos showed variation, the full-term developmental efficacy of these embryos were not different. These findings suggest that the methylation level of the satellite I sequence at the BC stage is not related to the abnormalities of bovine embryos produced by NT-SC. There was no difference in methylation levels between Re-NT and NT-SC embryos. Our results indicated that the DNA methylation status differed among embryos produced by various methods and that at least some of the demethylation of the donor cell genome occurred in the recipient cytoplast after NT-SC, but the demethylation ability of the NT procedure was noted in the first NT but not in the second NT.     

Recombinant canine granulocyte colony-stimulating factor accelerates recovery from cyclophosphamide-induced neutropenia in dogs

In dogs injected intravenously with 400mg/m(2) cyclophosphamide (CPA), the peripheral neutrophil count decreased to less than 1000 cells/μL in 5-9 days. Treatment with purified recombinant canine granulocyte colony-stimulating factor (rcG-CSF), produced by brevibacillus expression system, at the nadir of the granulocyte count accelerated recovery from the CPA-induced neutropenia by 1-3 days. Therapeutic administration of rcG-CSF at doses of 2.5-10 μg/kg did not show any significant difference on the severity of neutropenia (the period that granulocyte counts were less than 2000 cells/μL). Administration of 2.5 μg/kg rcG-CSF 3 times per day 2-4 days or 3-5 days after CPA treatment not only accelerated recovery but also decreased the severity of neutropenia. No clinical signs of the rcG-CSF were observed. These results showed that the rcG-CSF is effective for treatment of neutropenia in dogs.     

Identification of the strong vasorelaxing substance scirpusin B, a dimer of piceatannol, from passion fruit (Passiflora edulis) seeds

Piceatannol is present in passion fruit (Passiflora edulis) seeds in high amounts. In this study, we isolated the second major polyphenolic compound of passion fruit seeds and identified it as scirpusin B, which is a dimer of piceatannol. We investigated the antioxidant activities and vasorelaxing effects of these polyphenols. Their antioxidant effects were measured using an in vitro 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, and their vasorelaxant effects were determined ex vivo in rat thoracic aorta. Both polyphenolic compounds exhibited potent antioxidant activities and significant vasorelaxant effects in endothelium-intact aortas. More specifically, scirpusin B exerted a greater antioxidant activity and vasorelaxant effect compared with that of piceatannol. Additionally, the vasorelaxation effects of the compounds were induced via the NO derived from the endothelium. This study provides the possibility that polyphenols in passion fruit seeds are effective against cardiovascular diseases (CVDs).     

Establishment and characterization of a canine sebaceous epithelial cell line derived from an eyelid mass

Little is known about the pathological roles of sebaceous glands in canine skin diseases, as most examinations have been conducted with cultured human sebaceous epithelial cell lines. To our knowledge, there is no available canine sebaceous epithelial cell line. The purpose of this study was to establish a canine sebaceous epithelial cell line and characterize it. An eyelid mass in a dog was surgically resected for treatment, and it was histologically diagnosed as sebaceous epithelioma. Collected tissue was conducted for culture, and the growing epithelial-like cells were passaged. The cells showed continuous proliferation for over 6 months. After 40 passages, the cells were named CMG-1. Lipid droplets in the cytoplasm of CMG-1 cells were confirmed by Oil Red O staining. As reported in studies with human sebaceous epithelial cell lines, lipogenesis in CMG-1 cells was promoted by linoleic acid, whereas transforming growth factor-β (TGF-β) suppressed it. Additionally, real-time PCR revealed that the expression levels of chemokines and cytokines, including CC chemokine ligand (CCL)-2, CCL-20, CXCL-10, Tumor necrosis factor-α (TNF-α), Interleukin (IL)-1α, IL-1β, and IL-8, were significantly increased in CMG-1 cells following treatment with lipopolysaccharide. In conclusion, we successfully established a new canine sebaceous epithelial cell line. Our data indicated that lipogenesis and inflammatory responses were quantitatively evaluable in this cell line. CMG-1 cells could be useful for the pathological analysis of sebaceous gland diseases in dogs.     

Nutrient availability and response of sago palm (Metroxylon sago Rottb.) to controlled release N fertilizer on coastal lowland peat in the tropics

Sago palms (Metroxylon sagu Rottb.) growing on peat soils were found to grow more slowly and to show a lower production than palms growing on mineral soils. This difference was related to the physical and chemical constraints of peat soils, which include low bulk density, high acidity, and low N, P, K, Ca, Zn, and Cu levels. In coastal lowland peat soils, the distance from the sea has been found to be an important determinant of soil elemental composition. We predicted that a sufficient supply of N at the rosette stage would improve sago palm growth and that the availability of N in soil to which controlled release N fertilizer was applied might be higher than that in soil treated with soluble fertilizer. To investigate the changes in the nutrient composition of peat soils at various distances from the sea and the effect on sago palm growth, we studied sago palm areas in Indonesia and Malaysia. To observe the influence of N on the growth performance, we also conducted a fertilizer experiment on coastal lowland peat soil in Indonesia. Distance from the sea had no significant effect on the cation concentration in the soil solution (with the exception of Mg) or on the levels of soil-exchangeable cations. No significant differences were observed between the concentrations of exchangeable cations in surface peat soils and those in mature leaves. However, the concentrations of K, Na, and Ca in mature leaves increased significantly with their concentrations in the soil solution. This finding implies that the concentrations of cations in sago palm leaves depend directly on the concentrations of cations in the soil solution. No significant effect of N fertilizers on plant height and leaf formation was observed. N fertilizers applied twice a year did not affect appreciably the foliar concentration of N determined in December 1998 (5 months after the initial application) and December 1999. In June 2000, we detected a significantly higher concentration of N (p < 0.01) in young leaves of the palms treated with LP-100 or urea than in control leaves. However, no significant difference was detected between the LP-100 and urea treatments in the concentration of N in both mature and young leaves. This finding indicated that the concentration of N in sago palm leaves increased with the level of soil-applied N, regardless of whether N was applied as controlled release fertilizer or in the soluble form. We anticipate that a significant difference in the effects of these N fertilizers may occur during the next rainy season, when there should be a considerable loss of soluble N.     

Influence of surface printing materials on the degradability of biodegradable plastic films in soil

Effect of surface printing on the biodegradability of plastic films was studied. Biodegradable films (polybutylene-succinate (PBS)) printed with four kinds of gravure inks were placed in soil for 1 year. The inks consisted of carbon black-pigment with four kinds of resins: poly-(ε-caprolactone) (PCL), nitrocellulose-polyamide blended resin (NT), polyvinyl chloride-vinyl acetate copolymer (V), and nitrocellulose (NC). Degradation of film specimens printed on one side and both sides as well as the control film without printing was monitored every 3 months by collecting sample specimens for the measurement of weight loss. No appreciable degradation was observed until 6 months after placement in soil for the control specimens and until 9 months for the printed specimens. And the degradation of the PCL- and NC-printed specimens with one-side printing and V-printed specimens with both-side printing was significantly slower than that of the control specimens without printing after 9-month placement at p < 0.05. Only after 12 months of placement, was the degradation significantly faster for the specimens printed on one side than for those printed on both sides except of the specimens printed with NC. There was no difference in biodegradability among PCL, NT, NC, and V resins. Specimens printed on both sides did not show any appreciable weight loss after 1 year in soil (percentage of maintenance of weight exceeding 98%). Microscopic observation indicated that the degradation mainly proceeded from the non-printed side to the printed side cross-sectionally.     

Nutrient and bulk density characteristics of soil profiles in six land use systems along topo-sequences in inland valley watersheds of Ashanti region,Ghana

The bulk density and nutrient distribution in soil profiles as affected by various land use systems were studied. Mean bulk densities of the various soil profiles, 0-20, 20-40, and 4060 cm in the selected five land use systems along the upland / lowland topo-sequences were, 1.26, 1.73, and 1.44 Mg m^-3 for the primary forest (PF) plot in the upland area, 1.13, 1.51, and 1.50 Mg m^-3 for the cacao farm (CP) plot, 1.16, 1.63, and 1.26 Mg m^-3 for the mixed cropping (MC) plot, 1.39, 1.61, and 1.50 Mg m-a for the fallow (Fallow) plot, all three land use systems being in the upland / fringe areas, and 1.46, 1.72, and 1.60 Mg m^-3 for the lowland traditional rice farming (TR) plot, respectively. The very high bulk density of the second layer, especially in the PF plot, may be related to the hardening of the iron / quartz stone layer due to prolonged dry conditions, which may affect plant growth and hydrological cycles in the benchmark inland valley watersheds. The TC contents of the topsoil samples were 43, 32, 24, 26, 18, and 15 g kgm^-1 for the PF, CP, Fallow, MC, TR, and sawah (Sawah) plots, respectively. The C / N ratios of the topsoil samples were 10, 10, 9.8, 9.1, 8.8, and 8.8 for the CP, Fallow, MC, Sawah, TR, and PF plots, respectively, displaying a higher natural fertility, especially for the N supply in the PF plot in the upland area. The available phosphorus levels were low even for the topsoil samples, 1.8 to 3.4 mg kg^-1 for the upland soils while the lowland TR and Sawah plots recorded values of 4.4 and 4.9 mg kg^-1, respectively. The contents of exchangeable K, 0.1-0.4 cmole kg^-1, of the topsoil samples were in the range of those of normal inland valley soils in West Africa. In contrast to the content of available P, although the MC plot showed a depletion, the PF, Fallow, and CP plots showed a replenishment of available K through biological nutrient cycling. The levels of exchangeable Ca and Mg as well as eCEC, were also relatively high compared to those of mean inland valley soils in West Africa. The major component of eCEC was Ex. Ca. The levels of Ex. Ca in the topsoil samples were 11.1, 9.1, 7.8, 4.7, 5.1, and 3.9 cmol_c kg^-1 for the PF, CP, Fallow, MC, TR, and Sawah plots, respectively. The distinctive feature of Ex. Mg was its large distribution in the upper soil profiles in the CP plot, indicating that in the cacao farms, Ex. Mg was enriched, probably due to the high level of Mg in the leaf litter. The lowland soils, normally, should have been enriched in exchangeable bases such as Ca, K, and Mg, leading to a higher eCEC value and higher general fertility through the process of geological fertilization, Le., nutrient flows from upland to lowland areas, as in the case of lowland sawah in monsoon Asia. At the benchmark sites, however, the effects of the geological fertilization process were not evident. This is one of the most important characteristics of West African inland valley watersheds.     

Aluminum oxyhydroxide polymorphs and some micromorphological characteristics in sclerotium grains

The presence of aluminum oxyhydroxide and its micromorphological characteristics in sclerotium grains were examined by SEM, TEM, and EDX analyses. A spherical fraction with a diameter of 100 nm, which contained C and Al as the predominant elements, was recognized as the unit particle of the matrix of sclerotium grains. The Al polymorphs and the Si spherical structures found in the ignited grains were assumed to correspond to boehmite and opaline silica particles, respectively. SEMI analysis revealed the presence of a needle ball structure inside the hollow part of sclerotium grains. The relationship between the inner structure of sclerotium and biochemical processes of the host fungi was discussed.     

Methane production and its fate in paddy fields

Abstract

Oxidation of methane and total water soluble organic carbon (TOC) in the subsoil, which percolated from the plow layer, was investigated in a column experiment. The amounts of both methane and TOC in the leachate decreased by percolation in the subsoil.

Fe²⁺ percolated from the plow layer was nearly completely retained in the subsoil. The decomposition of methane and TOC in the subsoil was considered to result in the coupling with the formation of Fe²⁺. Methane was estimated to contribute ca. 19–21% to the total amount of Fe²⁺ formed in the subsoil by the organic materials in the leachate.