Panax genus belonging to a family of Araliaceae grow in Asia(9 species)and in North America(2 species).Especially Panax ginseng was listed in Chinese medical book,Shennong's Classic of Materia Medica approximately...Panax genus belonging to a family of Araliaceae grow in Asia(9 species)and in North America(2 species).Especially Panax ginseng was listed in Chinese medical book,Shennong's Classic of Materia Medica approximately 2 thousand years ago and Panax species are now one of the most important natural medicine.Since Panax species contain approximately 260 ginsenosides,its quality control and pharmacological movement of ginsenoside in body are not enough understanding.Monoclonal antibodies against ginsenosides were prepared and set up the enzyme linked immunosorbent assay system for the quality control of natural product.Furthermore,we developed Eastern blotting system using monoclonal antibodies resulted that protopanaxadiol and protopanaxatriol group ginsenosides can be separately stained by two corresponding monoclonal antibodies,respectively.It became evident that the ginseng slice was stained by Eastern blotting system.Histochemical staining of ginseng can make clear the ginsenoside-Rb1 distribution in cells and tissues.Double Eastern blotting system facilitated by two monoclonal antibodies like anti-ginsenoside-Rb1 and anti-ginsenoside-Rg1 monoclonal antibodies gives several information such as sugar number,structure,and qualitative/quantitative evaluation.Immunoaffinity column combined with monoclonal antibodies succeeded one-step isolation of ginsenoside and make it possible to prepare knockout extract of which only antigen molecule was removed suggesting the pharmacological and biological value of antigen molecule in the crude extract.展开更多
Microfold (M) cells are a kind of intestinal epithelial cell in the follicle-associated epithelium (FAE) of Peyer’s patches. They can transport antigens and microorganisms to lymphoid tissues. Bovine spongiform encep...Microfold (M) cells are a kind of intestinal epithelial cell in the follicle-associated epithelium (FAE) of Peyer’s patches. They can transport antigens and microorganisms to lymphoid tissues. Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative disorder in cattle. It is linked to variant Creutzfeldt-Jakob disease in humans. Although it is thought that M cells transport the BSE agent, the exact mechanism by which it crosses the intestinal barrier is not clear. We have bovine intestinal epithelial cell line (BIE cells), which can differentiate into the M cell type in vitro after stimulation, and which is able to transport the BSE agent. We show here that M cells are able to incorporate large numbers of PrP coated magnetic particles into intracellular vesicles, which we collected. The results of 2-DE show a specific protein associated with the PrP-coated particles, compared with non-coated particles. This protein was identified as aldolase A, a glycolytic pathway enzyme, using LC-MS/MS analysis. Aldolase A was synthesized and secreted by BIE cells, and increased during M cell differentiation. In the villi of the bovine intestine, aldolase A was detected on the surface of the epithelium and in the mucus droplet of goblet cells. In the FAE of bovine jejunal and ileal Peyer’s patches, aldolase A was localized on the surface and the apical part of the M cells. The binding of rbPrP to aldolase A was clearly detected and inhibited by pre-treatment of anti-aldolase A antibody. Aldolase A was co-stained with incorporated PrPSc in M-BIE cells. These results suggest that bovine M cells and goblet cells synthesize aldolase A, and that aldolase A may have the ability to bind PrP and associate with PrP in cellular vesicles. Therefore, aldolase A-positive M cells may play a key role in the invasion of BSE into the body.展开更多
Different vegetation types used for the extensive green roofs have characteristic physiological and morphological traits (e.g., C3, C4, or CAM photosynthesis, deciduous or evergreen). Several Sedum species are recogni...Different vegetation types used for the extensive green roofs have characteristic physiological and morphological traits (e.g., C3, C4, or CAM photosynthesis, deciduous or evergreen). Several Sedum species are recognized as “inducible CAM” type plants. These differences in the physiological and morphological traits have a considerable effect on the carbon sequestration in the green roofs. The objective of the present study was to quantify the carbon sequestration in several green roof plants during the first year after the construction of the green roofs and to clarify the relevance of the physiological and morphological traits to each plant’s ability to sequester carbon in its body using the growth analysis method. We used Zoysia matrella , Ophiopogon japonicus , and Sedum mexicanum species for the study wherein, S. mexicanum was assigned to the wet, dry, and non-irrigation treatments, and Z. matrella and O. japonicus only received the wet treatment. During the first year after the construction, carbon sequestration in the plants and the substrate of S. mexicanum was in the range of 276 to 364 g-C/m2/year, which was similar to that of O. japonicus and the finding of a previous study. In contrast, Z. matrella exhibited the highest carbon sequestration (670 g-C/m2/year), which is also expressed as the relative plant C-sequestration rate per whole-plant C-content (RGRc), because Z. matrella is a C4 plant and exhibits the highest net assimilation rate (NARc) of all species. Significant differences were not observed in RGRc , NARc , and RMF (root mass fraction) in S. mexicanum between the wet and dry treatments. These results suggest that in countries with high rainfall, a high frequency of irrigation has an insignificant effect on the physiological and morphological characteristics, and carbon sequestration in the Sedum green roofs.展开更多
Some Angelica species are used for medicinal purposes. In particular, the roots of Angelica acutiloba var. acutiloba and A. acutiloba var. sugiyamae, known as “Toki” and “Hokkai Toki”, respectively, are used as im...Some Angelica species are used for medicinal purposes. In particular, the roots of Angelica acutiloba var. acutiloba and A. acutiloba var. sugiyamae, known as “Toki” and “Hokkai Toki”, respectively, are used as important medicinal materials in traditional Japanese medicine. However, since these varieties have recently outcrossed with each other, it is difficult to determine whether the Japanese Angelica Root material used as a crude drug is the “pure” variety. In this study, we developed an efficient method to authenticate A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae from each other and from other Angelica species/varieties. The random amplified polymorphic DNA (RAPD) method efficiently discriminated each Angelica variety. A. acutiloba var. sugiyamae was identified via a characteristic fragment amplified by the decamer primer OPD-15. This fragment showed polymorphisms among Angelica species/varieties. The unique fragment derived from A. acutiloba var. sugiyamae was also found in one strain of A. acutiloba var. acutiloba, implying that this strain arose from outcrossing between A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae. This RAPD marker technique will be useful for practical and accurate authentication among A. acutiloba var. acutiloba, A. acutiloba var. sugiyamae, and their adulterants.展开更多
Japanese Angelica Root prepared from Angelica acutiloba var. acutiloba and A. acutiloba var. sugiyamae, known in Japan as “Toki” and “Hokkai Toki”, is an important crude drug used in Kampo medicine (traditional Ja...Japanese Angelica Root prepared from Angelica acutiloba var. acutiloba and A. acutiloba var. sugiyamae, known in Japan as “Toki” and “Hokkai Toki”, is an important crude drug used in Kampo medicine (traditional Japanese medicine). However, since these Angelica varieties have recently outcrossed with each other, it is unclear whether Japanese Angelica Root sold for use in Kampo medicine is a pure variety. Here, we describe DNA sequence polymorphisms that can be used to distinguish between A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae. In our analyses, differences in the trnK region of chloroplast DNA distinguished among some A. acutiloba varieties and related species, but not between A. acutiloba var. acutiloba and A. acutiloba var. iwatensis. One geographical strain of A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae showed identical sequences in three regions of chloroplast DNA, but differences in the internal transcribed spacer region of nuclear ribosomal DNA. One strain of A. acutiloba var. iwatensis and A. acutiloba var. sugiyamae had identical sequences in all of the chloroplast and nuclear ribosomal DNA regions examined. These findings show that A. acutiloba var. acutiloba has hybridized with A. acutiloba var. sugiyamae and that the “Hokkai Toki” variety resulted from outcrossing with A. acutiloba var. iwatensis. Molecular authentication based on analyses of chloroplast and nuclear ribosomal DNA sequences of A. acutiloba and related species is an efficient method to authenticate Japanese Angelica Root at the variety level. Therefore, these analyses can determine whether a product is derived from A. acutiloba var. acutiloba or A. acutiloba var. sugiyamae.展开更多
Caryopteris incana is a perennial shrub distributed in the temperate zone of the East Asia. It is found in West Kyushu in Japan, where it is designated as an endangered species. Tsushima, Nagasaki, which experienced r...Caryopteris incana is a perennial shrub distributed in the temperate zone of the East Asia. It is found in West Kyushu in Japan, where it is designated as an endangered species. Tsushima, Nagasaki, which experienced repeated connection and fragmentation between the Korean Peninsula and Japan, is an island on the route along which C. incana moved to Japan from continental Asia. We conducted field work and confirmed the genetic structure of populations using DNA sequence analysis to construct a detailed distribution map and clarify the intraspecific phylogenetic relationships of C. incana in Tsushima Island. We confirmed 72 populations in Tsushima. Using the leaves of individuals cultivated from seeds collected from each natural population, we analyzed the chloroplast and nuclear DNA sequence variations. Among the populations, sequence variations were confirmed in six regions of chloroplast DNA, and six haplotypes, including base substitutions, were distinguished. Two haplotypes were mainly divided at the border of the northern part of the southern island in Tsushima. One population in the northwestern part of the north island showed a haplotype derived from the southern part. This finding revealed that the distribution of C. incana had been artificially influenced. Several haplotypes were confirmed by sequence variations in the northern populations, but only one haplotype in the southern populations, suggesting that C. incana on the north island had separated early from the south island in Tsushima.展开更多
Caryopteris incana is a continental plant, transferred to Japan from continental Asia via a land bridge between the Korean Peninsula and Tsushima Islands during a glacial period. It currently grows wild in West Kyushu...Caryopteris incana is a continental plant, transferred to Japan from continental Asia via a land bridge between the Korean Peninsula and Tsushima Islands during a glacial period. It currently grows wild in West Kyushu, Japan. In a previous study, we investigated the distribution of C. incana in the Tsushima Islands and confirmed the genetic structure of populations by using chloroplast DNA sequence analysis, suggesting that different haplotypes were distributed in the same area. Thus, it seemed that populations of C. incana throughout the Tsushima Islands colonized at different times;each haplotype had remained within its population without mixing. In this study, we conducted fieldwork to construct a detailed distribution map in West Kyushu excluding the Tsushima Islands. Additionally, we confirmed genetic structure of the C. incana population in these areas by using chloroplast DNA sequence analysis to study the intraspecific phylogenetic relationship of C. incana in Japan. We confirmed 37 natural populations in 257 locations throughout West Kyushu excluding the 72 natural populations in the Tsushima Islands. We also confirmed a recent decreasing trend in the number of natural populations in the Nagasaki Mainland. Using the leaves of individuals cultivated from seeds collected from each natural population, we analyzed the chloroplast DNA sequence variations. Among the investigated populations, sequence variations were confirmed in six regions of chloroplast DNA, and those haplotypes were mainly classified into two groups distributed in different areas on the phylogenetic tree. This finding revealed that the common ancestor of C. incana in Japan diverged early into two groups, followed by a fragmentation in population distribution for each area. The haplotype network almost reflected the geographical distribution on haplotypes. However, several haplotypes that were distributed in other areas were confirmed in the Nagasaki Mainland, suggesting a complicated distribution formation in the past.展开更多
Half centuries of follow-up survey has enabled the architects and urban planners to design rationally by the aid of planning Nonetheless, limitation has occurred at planning because city has been changing its utility ...Half centuries of follow-up survey has enabled the architects and urban planners to design rationally by the aid of planning Nonetheless, limitation has occurred at planning because city has been changing its utility in accordance with its users＇ demand. In this paper, the authors proposed a method to analyze trait of users in market areas near stations by analyzing location based social network. After the datum collection from geotagged tweets, these GPS （global positioning system） datum were plotted to map attained from yahoo open location platform. Then the morphological analysis and terminology extraction system extracted the keywords and their scores. After calculating the distance from stations and users＇ GPS coordination, the authors extracted the array of keywords and corresponding scores in some station market area. Lastly, ratios of all users＇ scores and city＇s scores were calculated to examine the locality. Full combination of data collection, natural language processing and visualization enabled the authors to envisage distribution of collective background in city.展开更多
文摘Panax genus belonging to a family of Araliaceae grow in Asia(9 species)and in North America(2 species).Especially Panax ginseng was listed in Chinese medical book,Shennong's Classic of Materia Medica approximately 2 thousand years ago and Panax species are now one of the most important natural medicine.Since Panax species contain approximately 260 ginsenosides,its quality control and pharmacological movement of ginsenoside in body are not enough understanding.Monoclonal antibodies against ginsenosides were prepared and set up the enzyme linked immunosorbent assay system for the quality control of natural product.Furthermore,we developed Eastern blotting system using monoclonal antibodies resulted that protopanaxadiol and protopanaxatriol group ginsenosides can be separately stained by two corresponding monoclonal antibodies,respectively.It became evident that the ginseng slice was stained by Eastern blotting system.Histochemical staining of ginseng can make clear the ginsenoside-Rb1 distribution in cells and tissues.Double Eastern blotting system facilitated by two monoclonal antibodies like anti-ginsenoside-Rb1 and anti-ginsenoside-Rg1 monoclonal antibodies gives several information such as sugar number,structure,and qualitative/quantitative evaluation.Immunoaffinity column combined with monoclonal antibodies succeeded one-step isolation of ginsenoside and make it possible to prepare knockout extract of which only antigen molecule was removed suggesting the pharmacological and biological value of antigen molecule in the crude extract.
文摘Microfold (M) cells are a kind of intestinal epithelial cell in the follicle-associated epithelium (FAE) of Peyer’s patches. They can transport antigens and microorganisms to lymphoid tissues. Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative disorder in cattle. It is linked to variant Creutzfeldt-Jakob disease in humans. Although it is thought that M cells transport the BSE agent, the exact mechanism by which it crosses the intestinal barrier is not clear. We have bovine intestinal epithelial cell line (BIE cells), which can differentiate into the M cell type in vitro after stimulation, and which is able to transport the BSE agent. We show here that M cells are able to incorporate large numbers of PrP coated magnetic particles into intracellular vesicles, which we collected. The results of 2-DE show a specific protein associated with the PrP-coated particles, compared with non-coated particles. This protein was identified as aldolase A, a glycolytic pathway enzyme, using LC-MS/MS analysis. Aldolase A was synthesized and secreted by BIE cells, and increased during M cell differentiation. In the villi of the bovine intestine, aldolase A was detected on the surface of the epithelium and in the mucus droplet of goblet cells. In the FAE of bovine jejunal and ileal Peyer’s patches, aldolase A was localized on the surface and the apical part of the M cells. The binding of rbPrP to aldolase A was clearly detected and inhibited by pre-treatment of anti-aldolase A antibody. Aldolase A was co-stained with incorporated PrPSc in M-BIE cells. These results suggest that bovine M cells and goblet cells synthesize aldolase A, and that aldolase A may have the ability to bind PrP and associate with PrP in cellular vesicles. Therefore, aldolase A-positive M cells may play a key role in the invasion of BSE into the body.
文摘Different vegetation types used for the extensive green roofs have characteristic physiological and morphological traits (e.g., C3, C4, or CAM photosynthesis, deciduous or evergreen). Several Sedum species are recognized as “inducible CAM” type plants. These differences in the physiological and morphological traits have a considerable effect on the carbon sequestration in the green roofs. The objective of the present study was to quantify the carbon sequestration in several green roof plants during the first year after the construction of the green roofs and to clarify the relevance of the physiological and morphological traits to each plant’s ability to sequester carbon in its body using the growth analysis method. We used Zoysia matrella , Ophiopogon japonicus , and Sedum mexicanum species for the study wherein, S. mexicanum was assigned to the wet, dry, and non-irrigation treatments, and Z. matrella and O. japonicus only received the wet treatment. During the first year after the construction, carbon sequestration in the plants and the substrate of S. mexicanum was in the range of 276 to 364 g-C/m2/year, which was similar to that of O. japonicus and the finding of a previous study. In contrast, Z. matrella exhibited the highest carbon sequestration (670 g-C/m2/year), which is also expressed as the relative plant C-sequestration rate per whole-plant C-content (RGRc), because Z. matrella is a C4 plant and exhibits the highest net assimilation rate (NARc) of all species. Significant differences were not observed in RGRc , NARc , and RMF (root mass fraction) in S. mexicanum between the wet and dry treatments. These results suggest that in countries with high rainfall, a high frequency of irrigation has an insignificant effect on the physiological and morphological characteristics, and carbon sequestration in the Sedum green roofs.
文摘Some Angelica species are used for medicinal purposes. In particular, the roots of Angelica acutiloba var. acutiloba and A. acutiloba var. sugiyamae, known as “Toki” and “Hokkai Toki”, respectively, are used as important medicinal materials in traditional Japanese medicine. However, since these varieties have recently outcrossed with each other, it is difficult to determine whether the Japanese Angelica Root material used as a crude drug is the “pure” variety. In this study, we developed an efficient method to authenticate A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae from each other and from other Angelica species/varieties. The random amplified polymorphic DNA (RAPD) method efficiently discriminated each Angelica variety. A. acutiloba var. sugiyamae was identified via a characteristic fragment amplified by the decamer primer OPD-15. This fragment showed polymorphisms among Angelica species/varieties. The unique fragment derived from A. acutiloba var. sugiyamae was also found in one strain of A. acutiloba var. acutiloba, implying that this strain arose from outcrossing between A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae. This RAPD marker technique will be useful for practical and accurate authentication among A. acutiloba var. acutiloba, A. acutiloba var. sugiyamae, and their adulterants.
文摘Japanese Angelica Root prepared from Angelica acutiloba var. acutiloba and A. acutiloba var. sugiyamae, known in Japan as “Toki” and “Hokkai Toki”, is an important crude drug used in Kampo medicine (traditional Japanese medicine). However, since these Angelica varieties have recently outcrossed with each other, it is unclear whether Japanese Angelica Root sold for use in Kampo medicine is a pure variety. Here, we describe DNA sequence polymorphisms that can be used to distinguish between A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae. In our analyses, differences in the trnK region of chloroplast DNA distinguished among some A. acutiloba varieties and related species, but not between A. acutiloba var. acutiloba and A. acutiloba var. iwatensis. One geographical strain of A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae showed identical sequences in three regions of chloroplast DNA, but differences in the internal transcribed spacer region of nuclear ribosomal DNA. One strain of A. acutiloba var. iwatensis and A. acutiloba var. sugiyamae had identical sequences in all of the chloroplast and nuclear ribosomal DNA regions examined. These findings show that A. acutiloba var. acutiloba has hybridized with A. acutiloba var. sugiyamae and that the “Hokkai Toki” variety resulted from outcrossing with A. acutiloba var. iwatensis. Molecular authentication based on analyses of chloroplast and nuclear ribosomal DNA sequences of A. acutiloba and related species is an efficient method to authenticate Japanese Angelica Root at the variety level. Therefore, these analyses can determine whether a product is derived from A. acutiloba var. acutiloba or A. acutiloba var. sugiyamae.
文摘Caryopteris incana is a perennial shrub distributed in the temperate zone of the East Asia. It is found in West Kyushu in Japan, where it is designated as an endangered species. Tsushima, Nagasaki, which experienced repeated connection and fragmentation between the Korean Peninsula and Japan, is an island on the route along which C. incana moved to Japan from continental Asia. We conducted field work and confirmed the genetic structure of populations using DNA sequence analysis to construct a detailed distribution map and clarify the intraspecific phylogenetic relationships of C. incana in Tsushima Island. We confirmed 72 populations in Tsushima. Using the leaves of individuals cultivated from seeds collected from each natural population, we analyzed the chloroplast and nuclear DNA sequence variations. Among the populations, sequence variations were confirmed in six regions of chloroplast DNA, and six haplotypes, including base substitutions, were distinguished. Two haplotypes were mainly divided at the border of the northern part of the southern island in Tsushima. One population in the northwestern part of the north island showed a haplotype derived from the southern part. This finding revealed that the distribution of C. incana had been artificially influenced. Several haplotypes were confirmed by sequence variations in the northern populations, but only one haplotype in the southern populations, suggesting that C. incana on the north island had separated early from the south island in Tsushima.
文摘Caryopteris incana is a continental plant, transferred to Japan from continental Asia via a land bridge between the Korean Peninsula and Tsushima Islands during a glacial period. It currently grows wild in West Kyushu, Japan. In a previous study, we investigated the distribution of C. incana in the Tsushima Islands and confirmed the genetic structure of populations by using chloroplast DNA sequence analysis, suggesting that different haplotypes were distributed in the same area. Thus, it seemed that populations of C. incana throughout the Tsushima Islands colonized at different times;each haplotype had remained within its population without mixing. In this study, we conducted fieldwork to construct a detailed distribution map in West Kyushu excluding the Tsushima Islands. Additionally, we confirmed genetic structure of the C. incana population in these areas by using chloroplast DNA sequence analysis to study the intraspecific phylogenetic relationship of C. incana in Japan. We confirmed 37 natural populations in 257 locations throughout West Kyushu excluding the 72 natural populations in the Tsushima Islands. We also confirmed a recent decreasing trend in the number of natural populations in the Nagasaki Mainland. Using the leaves of individuals cultivated from seeds collected from each natural population, we analyzed the chloroplast DNA sequence variations. Among the investigated populations, sequence variations were confirmed in six regions of chloroplast DNA, and those haplotypes were mainly classified into two groups distributed in different areas on the phylogenetic tree. This finding revealed that the common ancestor of C. incana in Japan diverged early into two groups, followed by a fragmentation in population distribution for each area. The haplotype network almost reflected the geographical distribution on haplotypes. However, several haplotypes that were distributed in other areas were confirmed in the Nagasaki Mainland, suggesting a complicated distribution formation in the past.
文摘Half centuries of follow-up survey has enabled the architects and urban planners to design rationally by the aid of planning Nonetheless, limitation has occurred at planning because city has been changing its utility in accordance with its users＇ demand. In this paper, the authors proposed a method to analyze trait of users in market areas near stations by analyzing location based social network. After the datum collection from geotagged tweets, these GPS （global positioning system） datum were plotted to map attained from yahoo open location platform. Then the morphological analysis and terminology extraction system extracted the keywords and their scores. After calculating the distance from stations and users＇ GPS coordination, the authors extracted the array of keywords and corresponding scores in some station market area. Lastly, ratios of all users＇ scores and city＇s scores were calculated to examine the locality. Full combination of data collection, natural language processing and visualization enabled the authors to envisage distribution of collective background in city.
