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We have developed a quantitative determination method of the concentration of inorganic arsenic in pet foods using a liquid chromatograph-inductively coupled plasma-mass spectrometer (LC-ICP-MS). After adding 2 w/v% TMAH solution to a sample, inorganic arsenic was extracted by heating and the extract was collected by water. The pH of the solution was adjusted, and injected into a LC-ICP-MS to determine the concentration of inorganic arsenic. LC separation was carried out on an ODS column with 10 mmol/L sodium 1-butanesulfonate, 4 mmol/L malonic acid, 4 mmol/L TMAH and 0.05% methanol solution as a mobile phase. A collaborative study was conducted by nine laboratories using dry and wet-type pet foods, formed jerky, dried jerky and biscuit. Dry-type pet food and dried jerky was added with 2 mg/kg of As (III). Wet-type pet food was added with 0.5 mg/kg of As (III). Formed jerky was added with 1 mg/kg of As (III). Biscuit was added with 0.2 mg/kg of As (III). The mean recoveries, repeatabilities and reproducibilities in the form of relative standard deviation (RSDr and RSDR), and HorRat, were 95.4% to 98.3%, less than 2.9%, less than 9.1%, and 0.22 to 0.51, respectively.

Although variation among habitats in the ratio of gametophytes to sporophytes has been reported in various gigartinacean species, factors controlling the phase ratio remain poorly understood. Over 18 months, we examined the phase ratio of Chondrus ocellatus at three sites: a sheltered intertidal site, Hiruga A; an exposed intertidal site, Hiruga B; and a subtidal site, Shikimi. The mean proportion of gametophytes at Hiruga A (73.1%) was significantly higher than that at Shikimi (51.2%) and Hiruga B (44.7%). Due to a significantly higher water retention ability of the gametophytes, it was expected that the gametophytes would exhibit higher desiccation tolerance. After dehydration treatments, however, neither the photosynthetic rate of vegetative blades nor the survival rate of spores was significantly different between the phases. Measurements of blade strength indicated that the sporophytic blades were less stiff and more flexible, and a culture experiment revealed that the sporophytic germlings showed a significantly higher growth rate. Flexible blades and fast-growing germlings are considered advantageous for colonizing wave-swept intertidal habitats, so these properties may have caused the different fluctuation pattern of phase ratio among the sites. The present data demonstrate that biomechanical and physiological differences between the two phases of C. ocellatus make one phase advantageous in certain environmental conditions, and that these differences likely cause an unequal ratio of isomorphic phases.

The soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is a devastating pest of soybean (Glycine max (L.) Merr.) in the world. Three soybean QTLs for resistance to SCN race 1 were detected through QTL analyses using recombinant inbred lines (RILs) derived from a cross between 'Tokei 758' (susceptible) and 'To-8E' (resistant to races 1 and 3, derived from 'PI 84751' and 'Gedenshirazu'). Two of the three QTLs appear to be rhg1 and Rhg4 from their locations on the linkage map. The third QTL, detected around Satt359 on chromosome 11, was tentatively identified as rhg2. All RILs resistant to race 1 had all three QTLs. We developed lines carrying the three loci in various combinations, including all and none, from descendants of a cross between 'NIL-SCN' (with resistance derived from 'PI 84751' in the 'Natto-shoryu' background) and 'Natto-shoryu'. Evaluating these lines in a race 1-infected field in Mito, Ibaraki, showed that resistance to race 1 required all three loci. Through field evaluation of 10 recombinant fixed pairs that we developed, we located the rhg2 locus to an 821 kb-region between SSR markers Sat_123 (=WGSP11_0140) and BARCSOYSSR11_1420 on chromosome 11.

To clarify the effect of collagen addition to transplanted adipose tissue on angiogenesis, cell proliferation and tissue remodelling process and reveal whether collagen addition contributes to improving transplanted adipose tissue engraftment in rats. Adipose tissue was harvested from the inguinal and injected into the back of the rat, in addition to collagen. Engraftment tissue was harvested, semi-quantitatively evaluated and underwent haematoxylin and eosin or Perilipin staining. Moreover, we evaluated viable adipocyte counts and neovascularisation. Macrophages were evaluated using flow cytometry, and the adiponectin or vascular endothelial growth factor (VEGF) mRNA was detected using real-time polymerase chain reaction. By collagen addition to transplanted adipose tissue, higher engraftment rate semi-quantitatively and a greater number of new blood vessels histologically were identified. Perilipin staining revealed a higher adipocyte number. The total cell, M1 macrophage and M2 macrophage count were higher. There was increased adiponectin mRNA significantly at week 4 compared to that at week 1 after transplantation. Note that the expression levels of VEGF mRNA increased. In rats, adding collagen enhanced cell proliferation, induced M2 macrophages, which are involved in wound healing, and promoted adipocytes and neovascularisation. Therefore, collagen addition to transplanted adipose tissue could increase the engraftment rate of adipose tissue.

Objectives: To clarify the effect of impregnating transplanted adipose tissue with collagen on angiogenesis, cell proliferation, and tissue remodeling process and to reveal whether collagen impregnation contributes to improving the engraftment of transplanted adipose tissue in rats.Methods: Adipose tissue was harvested from the inguinal and injected into the back of the rat, in addition to collagen. Engraftment tissue was harvested, semi-quantitatively evaluated and underwent HE or Perilipin staining. Moreover, we evaluated viable adipocyte counts and neovascularization. Macrophages were evaluated using flow cytometry, and the adiponectin or VEGF mRNA was detected using real-time PCR. Results: By impregnating transplanted adipose tissue with collagen, higher engraftment rate semi-quantitatively and a greater number of new blood vessels histologically were identified. Perilipin staining revealed a higher adipocyte number. The total cell, M1 macrophage, and M2 macrophage count were higher. There was increased adiponectin mRNA significantly at week 4 compared to that at week 1 after transplantation. Note that the expression levels of VEGF mRNA increased.Discussion: In rats, collagen impregnation enhanced cell proliferation, induced M2 macrophages, which are involved in wound healing, and promoted adipocytes and neovascularization. Therefore, impregnating transplanted adipose tissue with collagen could increase the engraftment rate of adipose tissue.

Although variation among habitats in the ratio of gametophytes to sporophytes has been reported in various gigartinacean species, factors controlling the phase ratio remain poorly understood. Over 18 months, we examined the phase ratio of Chondrus ocellatus at three sites: a sheltered intertidal site, Hiruga A; an exposed intertidal site, Hiruga B; and a subtidal site, Shikimi. The mean proportion of gametophytes at Hiruga A (73.1%) was significantly higher than that at Shikimi (51.2%) and Hiruga B (44.7%). Due to a significantly higher water retention ability of the gametophytes, it was expected that the gametophytes would exhibit higher desiccation tolerance. After dehydration treatments, however, neither the photosynthetic rate of vegetative blades nor the survival rate of spores was significantly different between the phases. Measurements of blade strength indicated that the sporophytic blades were less stiff and more flexible, and a culture experiment revealed that the sporophytic germlings showed a significantly higher growth rate. Flexible blades and fast-growing germlings are considered advantageous for colonizing wave-swept intertidal habitats, so these properties may have caused the different fluctuation pattern of phase ratio among the sites. The present data demonstrate that biomechanical and physiological differences between the two phases of C. ocellatus make one phase advantageous in certain environmental conditions, and that these differences likely cause an unequal ratio of isomorphic phases.

The soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is a devastating pest of soybean (Glycine max (L.) Merr.) in the world. Three soybean QTLs for resistance to SCN race 1 were detected through QTL analyses using recombinant inbred lines (RILs) derived from a cross between ‘Tokei 758’ (susceptible) and ‘To-8E’ (resistant to races 1 and 3, derived from ‘PI 84751’ and ‘Gedenshirazu’). Two of the three QTLs appear to be rhg1 and Rhg4 from their locations on the linkage map. The third QTL, detected around Satt359 on chromosome 11, was tentatively identified as rhg2. All RILs resistant to race 1 had all three QTLs. We developed lines carrying the three loci in various combinations, including all and none, from descendants of a cross between ‘NIL-SCN’ (with resistance derived from ‘PI 84751’ in the ‘Natto-shoryu’ background) and ‘Natto-shoryu’. Evaluating these lines in a race 1-infected field in Mito, Ibaraki, showed that resistance to race 1 required all three loci. Through field evaluation of 10 recombinant fixed pairs that we developed, we located the rhg2 locus to an 821 kb-region between SSR markers Sat_123 (=WGSP11_0140) and BARCSOYSSR11_1420 on chromosome 11.

Radiocesium concentration in the crops cultivated in contaminated and decontaminated soils in the evacuation area were measured. The radiocesium concentration in the decontaminated soil decreased approximately one-quarter to that in the contaminated soil. The radiocesium concentration in the rice and vegetables cultivated in the decontaminated soil decreased to one-half to tenth of that in the contaminated soil. Except for eggplant and pumpkin cultivated in the decontaminated field, the radiocesium concentration in crops was higher than the standard limit in foods in Japan. The obtained transfer factors of most crops were within the reported values.

In Hokkaido, northern Japan, soybean [Glycine max (L.) Merr.] crops are damaged by cold weather. Chilling temperatures result in the appearance of cracking seeds (CS) in soybean crops, especially those grown in eastern and northern Hokkaido. Seed coats of CS are severely split on the dorsal side, and the cotyledons are exposed and frequently separated. CS occurrence causes unstable production because these seeds have no commodity value. However, little is known about the CS phenomenon. The aims of this study were to identify the cold-sensitive stage associated with CS occurrence and to develop a method to select CS-tolerant lines. First, we examined the relationship between chilling temperatures after flowering and CS occurrence in field tests. The average temperature 14 to 21 days after flowering was negatively correlated with the rate of CS. Second, we evaluated differences in CS tolerance among soybean cultivars and breeding lines in field tests. ‘Toyohomare’ and ‘Toiku-238’ were more CS-tolerant than ‘Yukihomare’ and ‘Toyomusume’. Third, we developed a selection method in which plants were subjected to 21-day chilling-temperature treatment from 10 days after flowering in a phytotron. This enabled comparisons of CS tolerance among cultivars. This selection method will be useful for breeding CS-tolerant soybeans.

In this study, we used an improved and more complex productmeter than previously designed for research and education. We performed daytime measurements on photosynthesis of Ulva pertusa on equipment to support one day, in order to examine its potential use as a teaching tool in university (higher education) programs to study environmental maintenance, reproduction, and creation along coastal areas. The productmeter is a tool used to measure the volume change in gas while maintaining a fixed concentration of gas in a container, which in turn, compresses a sample. In this study, we conducted an all day measurement using a complex productmeter; we intend to introduce this apparatus as a tool for higher education.

A better understanding of the relationships among different cropping systems, their effects on soil microbial ecology, and their effects on crop health and productivity is necessary for the development of more efficient, sustainable crop production systems. We used denaturing gradient gel electrophoresis (DGGE) to determine the impacts of crop rotations and crop types on bacterial and fungal communities in the soil. The communities of bacterial 16S rRNA genes and fungal 18S rRNA genes were analyzed in experimental field plots that were kept under 4 different crop rotation systems from 1999 to 2008 (continuous cabbage (Brassica oleracea var. capitata L.), cabbage–lettuce (Lactuca sativa L.) rotation, cabbage–radish (Raphanus sativus L. var. longipinnatus L.H. Bailey) rotation, and a 3-year crop rotation). A principal component analysis (PCA) and a canonical correspondence analysis (CCA) revealed that both the bacterial and fungal communities in bulk soils were influenced by the crop rotation systems. Howev...

Soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is one of the most damaging pests of soybean (Glycine max (L.) Merr.). Host plant resistance has been the most effective control method. Because of the spread of multiple SCN races in Hokkaido, the Tokachi Agricultural Experiment Station has bred soybeans for SCN resistance since 1953 by using 2 main resistance resources PI84751 (resistant to races 1 and 3) and Gedenshirazu (resistant to race 3). In this study, we investigated the genetic relationships of SCN resistance originating from major SCN resistance genes in Gedenshirazu and PI84751 by using SSR markers. We confirmed that race 1 resistance in PI84751 was independently controlled by 4 genes, 2 of which were rhg1 and Rhg4. We classified the PI84751- type allele of Rhg1 as rhg1-s and the Gedenshirazu-type allele of Rhg1 as rhg1-g. In the cross of the Gedenshirazu-derived race 3-resistant lines and the PI84751-derived races 1- and 3-resistant lines, the presence of rhg1-s and Rhg4 was responsible for race 1-resistance. These results indicated that it was possible to select race 1 resistant plants by using marker-assisted selection for the rhg1-s and Rhg4 alleles through a PI84751 origin × Gedenshirazu origin cross.

We simultaneously examined the bacteria, fungi and nematode communities in Andosols from four agro-geographical sites in Japan using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and statistical analyses to test the effects of environmental factors including soil properties on these communities depending on geographical sites. Statistical analyses such as Principal component analysis (PCA) and Redundancy analysis (RDA) revealed that the compositions of the three soil biota communities were strongly affected by geographical sites, which were in turn strongly associated with soil characteristics such as total C (TC), total N (TN), C/N ratio and annual mean soil temperature (ST). In particular, the TC, TN and C/N ratio had stronger effects on bacterial and fungal communities than on the nematode community. Additionally, two-way cluster analysis using the combined DGGE profile also indicated that all soil samples were classified into four clusters corresponding to the four sites, showing high site specificity of soil samples, and all DNA bands were classified into four clusters, showing the coexistence of specific DGGE bands of bacteria, fungi and nematodes in Andosol fields. The results of this study suggest that geography relative to soil properties has a simultaneous impact on soil microbial and nematode community compositions. This is the first combined profile analysis of bacteria, fungi and nematodes at different sites with agricultural Andosols.

1件の見逃しが大きなリスクを負う特許検索においては，検索結果の網羅性が重要である。皮肉なことに，問題特許ほど，検索の網に入りにくい言葉で書かれているため，検索技術がいくら向上しても，全文検索やテキスト検索には限界がある。この限界を超えるのは，特許分類の活用である。欧州特許分類は，欧州各国の特許を分類しているだけでなく，世界の特許を分類するツールとして使用されている。商用のデータベースに収録されたのは，まだ最近のことであるから，日本ではなじみが少ないが，欧州各国の特許検索に限らず，特に米国特許検索においても大きな威力を発揮する。欧州特許分類の創設の経緯から仕組み，国際特許分類との関係や今後の動向を明らかにし，実務における欧州特許分類の活用法を紹介する。

The number and distribution of branches in soybean plants influence seed yield through effects on the efficiency of light utilization as well as on tolerance to lodging. We have developed recombinant inbred lines (RILs) from a cross between two experimental determinant lines, which differ in branching number. The 172 RILs were divided into four maturity groups according to their alleles for two maturity loci, E1 and E3, and were planted separately to avoid confounding effects of competition. The late-maturity RIL groups with the E1 genotype were grown in two different locations, whereas the early-maturity RIL groups with the e1 genotype were planted at one location. Analysis of all lines resulted in the identification of five quantitative trait loci (QTLs) for branching number, designated qBr1 to qBr5. Among these QTLs, qBr1 and qBr2 were mapped to the proximal regions of the E1 and E3 loci, respectively. The other three QTLs were mapped to regions distant from any known maturity loci and were detected only in the presence of the E1 genotype, indicating that they interact with qBr1. Our results suggest that branching number might be controlled genetically by the identified QTLs, even though the maturity loci substantially affect branching phenotype.

The soil microbial community is strongly influenced by a wide variety of factors, such as soil characteristics and field management systems. In order to use biological indicators based on microbial community structure, it is very important to know whether or not these factors can be controlled. The present study aimed to determine whether soil type or fertilization has a greater influence on the soil microbial community based on denaturing gradient gel electrophoresis (DGGE) analysis of 12 experimental field plots containing four different soil types, Cumulic Andosol, Low-humic Andosol, Yellow Soil and Gray Lowland Soil, kept under three different fertilizer management systems since 2001 (the application of chemical fertilizer, the application of rice husk and cow manure, and the application of pig manure). Bacterial DGGE analysis using 16S rRNA genes and fungal DGGE analysis using 18S rRNA genes revealed that the bacterial community was related to the soil type more than the fertilization; howev...

北海道内全域で栽培されたダイズのイソフラボン含量を調査したところ, 同一品種でも栽培地によって含量は変動し, 登熟期間の平均気温(以下, 登熟気温)との間に有意な負の相関が認められた. また, 同一栽培地での明らかな品種間差も確認され, 「ゆきぴりか」および「音更大袖」の含量が最も高かった. ゆきぴりかは, 各適応地帯において標準品種(トヨコマチ)比1.3～1.5倍の高含量を示す一方, 栽培地間の変動係数は年次間変動を含めても標準品種より低かった. また, ゆきぴりかの高イソフラボンは, 登熟後半におけるダイズイン類の蓄積が一般品種より顕著であることに起因しており, ダイズイン類のゲニスチン類に対する比率(D/DG率)も高まっていたが, 品種間および交雑後代系統間にイソフラボン含量とD/DG率の有意な相関は認められず, これらは, 独立した遺伝形質であると推察された. 一方, D/DG率は同一品種でも栽培地によって変動し, イソフラボン含量と同様に登熟気温と有意な負の相関が見られたことから, イソフラボン蓄積を促進する条件は, D/DG率の決定にも何らかの影響を及ぼしていると考えられた.

In general, red tide indicates a visual and customary name for the "phenomenon that microalgae in seawater proliferate abnormally, causing a sudden change in sea surface color." However, this is not sufficient to quantitatively understand red tide, and the definition of red tide is not clear. On the other hand, the Ise Bay is considered to have eutrophication and red tide is regularly observed in the Bay. Although the load of nutrient salts has been decreasing, the Bay is one of the areas where determining the relation between red tide and water quality control is difficult. In this study, the author established the criteria to define red tide (red tide index), applied a stepwise method to a multiple regression model, and determined a stable model that has Chl-a (red tide index) as its explanatory variable. Environmental influence was captured by the change in the coefficient of determination of the red tide index model, which implicitly indicated the influence of the Fifth Regulation of Total Pollutant Load Control.

CO2 dissolving experiments in marine areas may cause some negative impact including changes in pH levels in the surrounding environment. Based on a CO2 dissolving experiment in both human waste water as well as artificial seawater using microbubbles, we conducted a basic study on a seaweed bed in Eutrophic Regions assuming CO2 fixation. if we check CO2 levels and operating time to avoid rapid elevation of pH levels in sewage treatment facilities, thus having the advantages of both techniques.

Tokyo Bay, Ise Bay, and the Seto Inland Sea are the total pollutant load control target areas in Japan. A significant correlation between the incidence of red tides and water quality has been observed in the Seto Inland Sea (Honjo, 1991). However, while red tides also occur in Ise Bay and Tokyo Bay, similar correlations have not been observed. Hence, it is necessary to understand what factors cause red tides to effectively manage these semi-closed systems. This study aims to investigate the relationship between the dynamics of the Red Tide Index and nitrogen regulation as well as phosphorus regulation, even in Ise Bay where, unlike Tokyo Bay, there are few observation items, by selecting a suitable objective variable. The introduction of a new technique that uses the Red Tide Index has revealed a possibility that the total pollution load control has influenced the dynamics of red tide blooms in Ise Bay.

A tissue kallikrein was purified over 1500-fold from the postmicrosomal supernatant of human submaxillary glands. The purified enzyme gave a single band, corresponding to an apparent molecular weight of 42,000 on SDS-polyacrylamide gel electrophoresis. This enzyme cross-reacted with the anti-human urinary kallikrein antiserum. The purified enzyme was characterized in comparison with the purest human urinary kallikrein preparation. Both enzymes hydrolyzed the synthetic substrate, Ac-Phe-Arg-OMe, most effectively. Aprotinin, TLCK, and PMSF suppressed the enzyme activities, while SBTI, LBTI, and alpha 1-antitrypsin had no effect at all. The purified enzyme generated kinin from the natural substrate, kininogen. It was concluded therefore that the purified enzyme is a typical tissue kallikrein.