Evaluation of alternative substrates for determining methane-oxidizing activities and methanotrophic populations in soils

The magnitude of methane emission is a net result of methane production and the oxidation rate. The possibility of measuring oxidized products of alternative substrates of methane monooxygenase was examined to determine methane-oxidizing ability of soils, and to count methanotrophic populations in soils. Wetland rice soils were incubated under methane containing air to enirch the methanotrophs. Methane loss and oxygen uptake were inhibited by acetylene, dimethylether, and nitrapyrin (N-Serve). Acetylene was used routinely, because it inhibited methane oxidation even at a low concentration of 0.03 to 0.06 l ml^-1 in the incubation headspace. Propylene at 10 kPa was used as an alternative substrate of methane monooxygenase, and the formation of propylene oxide was measured. When soils were incubated under methane, their methane-oxidizing activity increased. Propylene oxide formation increased simultaneously. Acetylene also blocked propylene oxidation. The results of several experiments and propylene oxide formation (r=0.87 after long-transformation). These results indicate that propylene oxide formation can be used as a semiquantitative measure of the methane-oxidizing activity of soils. The colonies of soluble methane monooxygenase-forming methanotrophs were counted on Cu-deficient methanotroph agar medium by the formation of naphthol from haphthalene. The counts increased from 10⁴ (0 days) to 10⁷ (21 days) g^-1 soil during oxic incubation under methane.     

Effects of N sources and methane concentrations on methane uptake potential of a typical coniferous forest and its adjacent orchard soil

After removal of the above-ground plant debris three different soil layers were taken from a typical coniferous forest and its adjacent orchard in Numata City, Japan. The potentials of soil CH₄ uptake at two initial CH₄ concentrations were studied under aerobic conditions in the laboratory, along with inhibition of soil CH₄ oxidation by urea-N or KNO₃-N addition. Due to long-term N inputs, the CH₄ uptake of the upper mineral layer of the orchard soil was 25.4% and 87.7% lower than that of the surface forest soil at 2.4 and 12.6 l l^–1 CH₄, respectively. Methane uptake of the forest soil decreased with increasing soil depths at two CH₄ levels. However, maximal CH₄-consuming activity occurred in the 9- to 23-cm depth of the orchard soil at 12.6 l l^–1 methane. Nitrogen additions in the form of KNO₃ or urea at the rate of 200 g N g^–1 soil substantially reduced soil CH₄ uptake in the upper and sub-surface mineral layers at both sites, except that the addition of KNO₃-N had no apparent inhibitory effect on the CH₄ uptake in the 9- to 23-cm depth of the orchard soil. A strong inhibitory effect of NO₃^– addition on the CH₄ uptake, in contrast to NH₄⁺, occurred in the surface forest soil. The use of KNO₃-N, as compared to urea or urea plus a nitrification inhibitor (dicyandiamide), resulted in a lower potential to cause inhibition of CH₄ oxidation in the 0- to 23-cm depth of the orchard soil.     

Biological dinitrogen fixation and soil microbial biomass carbon as influenced by free-air carbon dioxide enrichment (FACE) at three levels of nitrogen fertilization in a paddy field

An experiment was conducted in an Andosol paddy field in Shizukuishi (Iwate Prefecture, Japan) to determine the effects of free-air CO₂ enrichment (FACE) on biological N₂-fixation activity and soil microbial biomass C at three levels of N application. Rice (Oryza sativa L. cv. Akitakomachi) plants were grown under ambient CO₂ or FACE (ambient +200 µmol mol^-1 CO₂) conditions throughout the growing season with each treatment having four replicated plots. Three levels of N fertilizer (high, standard and low; 15, 9 and 4 g N m^-2, respectively) were applied to examine the effect of different N availability under both CO₂ conditions. Soil samples were collected at four different times from upper and lower soil layers (0-1-cm and 1-10-cm soil depths, respectively) and analysed for biological N₂-fixation (BNF) activity and microbial biomass C (MBC) by the acetylene reduction and chloroform fumigation-extraction methods, respectively. The amounts of chlorophyll-type compounds (Chls), an index of algal growth, and soil available C were also determined. Compared to the ambient CO₂ treatment, the FACE treatment had significantly higher BNF activity in both the upper and lower soil layers at ripening only in low-N soil and at harvest at all three levels of N fertilization rates. MBC was significantly increased by FACE in both the upper and lower soil layers from the middle to later period of the growing season compared to the ambient CO₂ treatment. The FACE treatment increased the Chls in the upper soil layers at ripening only in low-N soil and at harvest at all three levels of N fertilization rates. The amount of soil available C was not significantly different between FACE and ambient CO₂ treatments in both the upper and lower soil layers throughout the cropping season. From these results it can be concluded that the FACE treatment had a significantly positive influence on BNF activity, MBC and Chls at different levels of N fertilization rates in paddy field during the cropping season.     

Development of a multiplex PCR method for simultaneous detection of diagnostic DNA markers of five disease and pest resistance genes in potato

Multiplex PCR is practically a reasonable choice for molecular marker-assisted selection in potato breeding. We had developed and were using a multiplex PCR method for selection of resistance genes to cyst nematode (H1), Potato virus X (Rx1) and late blight (R1 and R2). Since then, more reliable and tightly linked markers for H1 and R2, and a new marker for resistance to Potato virus Y (Ry _chc ) were developed. In this article, all these superior markers, including a positive marker to eliminate PCR-failed samples, were incorporated into one multiplex PCR assay. Using the newly developed multiplex PCR technique, five plants potentially harboring all five resistance genes were selected from 96 hybrid plants approximately 5 h after DNA extraction, which is a third of the operation time compared with separate PCR reactions for each marker.     

Protostellar feedback halts the growth of the first stars in the universe

The first stars fundamentally transformed the early universe by emitting the first light and by producing the first heavy elements. These effects were predetermined by the mass distribution of the first stars, which is thought to have been fixed by a complex interplay of gas accretion and protostellar radiation. We performed radiation-hydrodynamics simulations that followed the growth of a primordial protostar through to the early stages as a star with thermonuclear burning. The circumstellar accretion disk was evaporated by ultraviolet radiation from the star when its mass was 43 times that of the Sun. Such massive primordial stars, in contrast to the often-postulated extremely massive stars, may help explain the fact that there are no signatures of the pair-instability supernovae in abundance patterns of metal-poor stars in our galaxy.     

Oxidative activation of O,S-dimethyl phosphoramidothiolate

O,S-Dimethyl phosphoramidothiolate, the insecticide methamidophos, is converted oxidatively into a potent anti-cholinesterase agent by the action of m-chloroperbenzoic acid. The active intermediate is not stable enough to be isolated. From some chemical evidence, the sulfoxide, i.e., dimethyl phosphoramidothiolate S-oxide, is most probably the active intermediate. A similar activation may occur by biological oxidation.     

Growth characteristics of canine distemper virus in a new cell line CCT cells originated from canine malignant histiocytosis

Canine distemper virus (CDV) growth and the morphological characterization were examined in a cell line established from a canine malignant histiocytosis (CCT cell line). The susceptibility of the CCT cells to 3 CDV strains, FXNO, YSA-TC and MD-77 was shown by detection of the antigen in the indirect fluorescent assay. After passaging 4 and 9 times through the CCT cells, only FXNO strain could produce the syncytia where demonstrated the antigens. Titers of 9 passaged viruses through the CCT cells showed slightly higher in the CCT cells than those in Vero cells. Morphological characterization of karyorrhexis and specific DNA ladder by extracted DNA electrophoresis indicated apoptosis in the CDV infected CCT cells.     

稻季施肥管理措施对后续麦季N2O排放的影响

2003─2004年选用江苏省宜兴市稻-麦轮作试验田,研究了水稻生长季秸杆(0和3.75×103kg/hm2两个水平)和N肥施用(N0、200和270kg/hm23个水平)对后续麦季N2O排放的影响。结果表明:稻季秸杆施用显著减少了后续麦季N2O的排放,这些减少量主要体现在小麦播种-返青期,方差分析达显著水平(P<0.05)。稻季施用N肥,后续麦季N2O排放减少,但N200和270kg/hm2N肥施用水平的处理间无显著差异。麦季土壤水分情况与N2O排放通量存在显著正相关(P<0.05)。     

Significance of tumor-infiltrating immune cells in spontaneous canine mammary gland tumor: 140 cases

The numbers of tumor infiltrating T lymphocytes, B lymphocytes and antigen presenting cells were evaluated in an immunohistochemical manner in 140 canine spontaneous mammary gland tumor (MGT) tissues. As a result, we found a statistically significant increase in the number of intratumoral T lymphocytes (23.2 ± 23.8) in the malignant MGT group (n=51) compared with the benign MGT group (14.0 ± 16.0, n=89; P<0.05). Moreover, the high T lymphocyte infiltration in the malignant group correlated with poor prognosis in multivariate analysis (P<0.05). This study indicated the relationship between increased infiltrating T lymphocytes and canine MGT malignancy.