K. Hamana, Ryuichi Hosoya, M. Niitsu, H. Hayashi, T. Itoh
Cellular polyamines acid-extracted from fourteen archaea (archaebacteria) belonging to the phyla, Euryarchaeota and Crenarchaeota, including nine newly validated species, grown under different culture conditions were analyzed using HPLC and GC. The following archaebacteria were studied: moderately thermophilic Methanothermobacter, Ferroplasma, Thermogymnomonas, and Acidianus species; a slightly thermophilic methanogen, Methanolinea tarda, and a mesophilic methanogen, Methanosaeta contilii, contained triamines and tetra-amines. Hyperthermophilic Thermococcus, Pyrococcus, and Ignicoccus contained a quaternary branched penta-amine, N4-bis(aminopropyl)spermidine, and two linear penta-amines, in addition to triamines and tetra-amines, respectively. When Thermococcus, Pyrococcus, and Sulfurisphaera species were cultivated under different growth temperatures (60-100°C) and salt concentrations (0.15%-2.5% NaCl), temperature and salt dependent long or branched polyamine syntheses, increasing under higher temperatures and higher salt conditions, were observed.
{"title":"Temperature and salt dependent long linear and branched polyamine syntheses in the thermophiles belonging to the domain Archaea","authors":"K. Hamana, Ryuichi Hosoya, M. Niitsu, H. Hayashi, T. Itoh","doi":"10.3118/JJSE.7.1.21","DOIUrl":"https://doi.org/10.3118/JJSE.7.1.21","url":null,"abstract":"Cellular polyamines acid-extracted from fourteen archaea (archaebacteria) belonging to the phyla, Euryarchaeota and Crenarchaeota, including nine newly validated species, grown under different culture conditions were analyzed using HPLC and GC. The following archaebacteria were studied: moderately thermophilic Methanothermobacter, Ferroplasma, Thermogymnomonas, and Acidianus species; a slightly thermophilic methanogen, Methanolinea tarda, and a mesophilic methanogen, Methanosaeta contilii, contained triamines and tetra-amines. Hyperthermophilic Thermococcus, Pyrococcus, and Ignicoccus contained a quaternary branched penta-amine, N4-bis(aminopropyl)spermidine, and two linear penta-amines, in addition to triamines and tetra-amines, respectively. When Thermococcus, Pyrococcus, and Sulfurisphaera species were cultivated under different growth temperatures (60-100°C) and salt concentrations (0.15%-2.5% NaCl), temperature and salt dependent long or branched polyamine syntheses, increasing under higher temperatures and higher salt conditions, were observed.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133464805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Wataru Tsukimura, K. Watanabe, C. Morokuma, R. Yatsunami, T. Fukui, Satoshi Nakamura
Xylanase B (XynTB) from hyperthermophilic bacterium Thermotoga maritima MSB8 is a thermostable xylanase classified into glycoside hydrolase family 10. XynTB is most active at pH 6.0, and shows lower activity at alkaline pHs. Improvement of alkaliphily of XynTB was attempted by directed evolution. One mutant enzyme that showed slightly higher activity under high temperature and alkaline pH conditions was acquired from a newly constructed random mutant library. Protein engineering study of this mutant revealed that the amino acid substitution N92D (Asn92 was substituted by Asp) could contribute to the improvement of alkaliphily.
{"title":"Improvement of alkaliphily of thermostable GH family 10 xylanase from Thermotoga maritima by directed evolution","authors":"Wataru Tsukimura, K. Watanabe, C. Morokuma, R. Yatsunami, T. Fukui, Satoshi Nakamura","doi":"10.3118/JJSE.9.15","DOIUrl":"https://doi.org/10.3118/JJSE.9.15","url":null,"abstract":"Xylanase B (XynTB) from hyperthermophilic bacterium Thermotoga maritima MSB8 is a thermostable xylanase classified into glycoside hydrolase family 10. XynTB is most active at pH 6.0, and shows lower activity at alkaline pHs. Improvement of alkaliphily of XynTB was attempted by directed evolution. One mutant enzyme that showed slightly higher activity under high temperature and alkaline pH conditions was acquired from a newly constructed random mutant library. Protein engineering study of this mutant revealed that the amino acid substitution N92D (Asn92 was substituted by Asp) could contribute to the improvement of alkaliphily.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"14 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115059284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The growing body of research reveal that the programmed cell death (PCD) is ubiquitously distributed in microorganisms, plays key roles in developmental processes and inkeeping the integrity of bacterial communities. Inspired by this, bioengineers are making their original version of PCD devices for the temporal and/or spatial control of the bacteria communities. In this review, we discuss how PCD system should be designed and/or implemented into the bacteria to make a faithful and long-lasting &39;robots&39; for the real-world applications.
{"title":"Device genetics for programmed cell death","authors":"Y. Tashiro, M. Kobayashi","doi":"10.3118/JJSE.7.2.31","DOIUrl":"https://doi.org/10.3118/JJSE.7.2.31","url":null,"abstract":"The growing body of research reveal that the programmed cell death (PCD) is ubiquitously distributed in microorganisms, plays key roles in developmental processes and inkeeping the integrity of bacterial communities. Inspired by this, bioengineers are making their original version of PCD devices for the temporal and/or spatial control of the bacteria communities. In this review, we discuss how PCD system should be designed and/or implemented into the bacteria to make a faithful and long-lasting &39;robots&39; for the real-world applications.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"91 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123125916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
G. Takayama, T. Kosuge, S. Sunamura, I. Matsui, K. Ishikawa, A. Nakamura, T. Hoshino
Genes annotated to threonine dehydrogenase, α-mannosidase, and glutamate dehydrogenase of the hyperthermophilic archaeon Pyrococcus horikoshii OT3 were used to test the expression system of the thermophilic eubacterium Thermus thermophilus HB27. Using the previously described P215 promoter, the three genes were successfully expressed, but at significantly lower levels than in the Escherichia coli expression system with the T7 promoter. Replacement of the promoter region with P31 or Pslp_promoter improved the expression to a level comparable to or exceeding that in the E. coli system. Notably, α-mannosidase activity was clearly detected with the P31 and Pslp_promoters, which could not be detected in the E. coli system. Moreover, with 6 x His-tag fusions of these enzymes at their COOH- or NH2-termini, these proteins could be detected in the crude extracts of T. thermophilus by Western blotting, indicating that the increment of each enzyme activity was actually the result of enzyme production. These results demonstrate that the host-vector system of T. thermophilus is useful for expression of genes from hyperthermophiles.
{"title":"Use of a Thermus thermophilus host-vector system for expression of genes from the hyperthermophilic archaeon Pyrococcus horikoshii","authors":"G. Takayama, T. Kosuge, S. Sunamura, I. Matsui, K. Ishikawa, A. Nakamura, T. Hoshino","doi":"10.3118/JJSE.3.1_28","DOIUrl":"https://doi.org/10.3118/JJSE.3.1_28","url":null,"abstract":"Genes annotated to threonine dehydrogenase, α-mannosidase, and glutamate dehydrogenase of the hyperthermophilic archaeon Pyrococcus horikoshii OT3 were used to test the expression system of the thermophilic eubacterium Thermus thermophilus HB27. Using the previously described P215 promoter, the three genes were successfully expressed, but at significantly lower levels than in the Escherichia coli expression system with the T7 promoter. Replacement of the promoter region with P31 or Pslp_promoter improved the expression to a level comparable to or exceeding that in the E. coli system. Notably, α-mannosidase activity was clearly detected with the P31 and Pslp_promoters, which could not be detected in the E. coli system. Moreover, with 6 x His-tag fusions of these enzymes at their COOH- or NH2-termini, these proteins could be detected in the crude extracts of T. thermophilus by Western blotting, indicating that the increment of each enzyme activity was actually the result of enzyme production. These results demonstrate that the host-vector system of T. thermophilus is useful for expression of genes from hyperthermophiles.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"12 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124231838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
One of the widest ranging abiotic stresses in world agriculture arises from low iron (Fe) availability due to high soil pH, with 30% of arable land too alkaline for optimal crop production. To aquire Fe, Graminaceous plants use a chelation strategy and release phytosiderophores from their roots to chelate Fe (III) in the soil. Non-graminaceous plants differ from graminaceous plants in acquiring Fe, ferric reduction on root surface being the first step of iron uptake. Rice is very susceptible to low Fe availability. We enhanced the tolerance of rice to Fe deficiency in calcareous soils by introducing the barley genes participating in the phytosiderophore synthesis and tested these transgenic rice lines in a field experiment on a calcareous soil under paddy conditions. We showed that introducing barley genes involved in the synthesis of phytosiderophore into rice is an effective, practical method to improve agricultural productivity in calcareous soils. We also improved the tolerance of rice to Fe deficiency by increasing the ferric reductase activity of root cells by introducing the engineered gene refre1-372. To understand the molecular mechanisms that regulate Fe acquisition in plants, we analyzed the promoter region of the barley Fe deficiency inducible IDS2 gene and identified two novel cis-acting elements, IDE1 and IDE2. We identified two rice transcription factors, IDEF1 and IDEF2, which specifically bind to IDE1 and IDE2, respectively. The enhanced expression of transcription factors resulted in increased phytosiderophore secretion and tolerance to low Fe availability in a calcareous soil.
{"title":"Enhancing tolerance of rice to alkaline soils using genes involved in Fe acquisition in plants.","authors":"N. Nishizawa","doi":"10.3118/JJSE.7.2.15","DOIUrl":"https://doi.org/10.3118/JJSE.7.2.15","url":null,"abstract":"One of the widest ranging abiotic stresses in world agriculture arises from low iron (Fe) availability due to high soil pH, with 30% of arable land too alkaline for optimal crop production. To aquire Fe, Graminaceous plants use a chelation strategy and release phytosiderophores from their roots to chelate Fe (III) in the soil. Non-graminaceous plants differ from graminaceous plants in acquiring Fe, ferric reduction on root surface being the first step of iron uptake. Rice is very susceptible to low Fe availability. We enhanced the tolerance of rice to Fe deficiency in calcareous soils by introducing the barley genes participating in the phytosiderophore synthesis and tested these transgenic rice lines in a field experiment on a calcareous soil under paddy conditions. We showed that introducing barley genes involved in the synthesis of phytosiderophore into rice is an effective, practical method to improve agricultural productivity in calcareous soils. We also improved the tolerance of rice to Fe deficiency by increasing the ferric reductase activity of root cells by introducing the engineered gene refre1-372. To understand the molecular mechanisms that regulate Fe acquisition in plants, we analyzed the promoter region of the barley Fe deficiency inducible IDS2 gene and identified two novel cis-acting elements, IDE1 and IDE2. We identified two rice transcription factors, IDEF1 and IDEF2, which specifically bind to IDE1 and IDE2, respectively. The enhanced expression of transcription factors resulted in increased phytosiderophore secretion and tolerance to low Fe availability in a calcareous soil.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"41 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124617833","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Y. Nishiguchi, T. Miwa, S. Kubota, M. Taru, M. Okada
A new zoarcid fish, Andriashevia natsushimae (Japanese name: Natsushimachojyagenge), is described on the basis of 2 specimens collected in Sagami Bay, Japan, at the depth of 850 m. This species has no pectoral fins and no pelvic fins, indicating it to be Andriashevia. It has a round body and a moderate-sized mouth in contrast to Andriashevia aptera which has a flat body and a large mouth. Phylogenic tree based on 16S ribosomal RNA (16S rRNA) sequence indicates the species had diverted at the early stage in the evolution of eelpouts.
{"title":"Characterization of Andriashevia natsushimae, a new species of eelpout (Pisces, Perciformes: Zoarcidae) from Sagami Bay, Japan, and its phylogenic status as inferred from 16S rRNA","authors":"Y. Nishiguchi, T. Miwa, S. Kubota, M. Taru, M. Okada","doi":"10.3118/JJSE.8.20","DOIUrl":"https://doi.org/10.3118/JJSE.8.20","url":null,"abstract":"A new zoarcid fish, Andriashevia natsushimae (Japanese name: Natsushimachojyagenge), is described on the basis of 2 specimens collected in Sagami Bay, Japan, at the depth of 850 m. This species has no pectoral fins and no pelvic fins, indicating it to be Andriashevia. It has a round body and a moderate-sized mouth in contrast to Andriashevia aptera which has a flat body and a large mouth. Phylogenic tree based on 16S ribosomal RNA (16S rRNA) sequence indicates the species had diverted at the early stage in the evolution of eelpouts.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"44 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123090048","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H. Tamegai, S. Nakamura, M. Miyazaki, Y. Nogi, R. Kasahara, C. Kato, K. Horikoshi
In the present study, we investigated the physiological properties of Pseudomonas sp. strain MT-1 isolated from the mud of Mariana Trench. Strain MT-1 was closely related with members of the genus Pseudomonas, especially with Pseudomonas chloritidismutans and Pseudomonas stutzeri on the basis of 16S rDNA sequence. The DNA-DNA hybridization values between strain MT-1 and Pseudomonas reference strains were significantly lower than those accepted as the phylogenetic definition of a species. MT-1 had polar flagellum, and was facultative anaerobe. The growth occurred in an NaCl concentration of about 0-10% (optimum: 1-2%), in pH of about 6-10 (optimum: 7-8), and in temperature of about 4-45 °C (optimum: 32-35 °C). The G+C content of the DNA was 60.5% mol%. The major quinone was ubiquinone-9. The major fatty acid in strain MT-1 was C16:0 (hexadecanoic acid), C16:1 (hexadecenoic acid) and C18:1 (octadecenoic acid). The organism showed adaptational properties to deep-sea environment compared with the reference strains.
{"title":"Physiological properties of Pseudomonas sp. strain MT-1, denitrifier from the 11,000 m-depth of Mariana Trench","authors":"H. Tamegai, S. Nakamura, M. Miyazaki, Y. Nogi, R. Kasahara, C. Kato, K. Horikoshi","doi":"10.3118/JJSE.4.25","DOIUrl":"https://doi.org/10.3118/JJSE.4.25","url":null,"abstract":"In the present study, we investigated the physiological properties of Pseudomonas sp. strain MT-1 isolated from the mud of Mariana Trench. Strain MT-1 was closely related with members of the genus Pseudomonas, especially with Pseudomonas chloritidismutans and Pseudomonas stutzeri on the basis of 16S rDNA sequence. The DNA-DNA hybridization values between strain MT-1 and Pseudomonas reference strains were significantly lower than those accepted as the phylogenetic definition of a species. MT-1 had polar flagellum, and was facultative anaerobe. The growth occurred in an NaCl concentration of about 0-10% (optimum: 1-2%), in pH of about 6-10 (optimum: 7-8), and in temperature of about 4-45 °C (optimum: 32-35 °C). The G+C content of the DNA was 60.5% mol%. The major quinone was ubiquinone-9. The major fatty acid in strain MT-1 was C16:0 (hexadecanoic acid), C16:1 (hexadecenoic acid) and C18:1 (octadecenoic acid). The organism showed adaptational properties to deep-sea environment compared with the reference strains.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"9 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116164808","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Dense animal communities at hydrothermal vents and cold seeps rely on symbioses with chemoautotrophic bacteria. To reveal the symbiotic mechanism, we sequenced the genome of chemoautotrophic intracellular symbiont in deep-sea clam, Calyptogena okutanii. On the other hand, C. magnifica symbiont genome sequenced in USA. The genomes appear to have been reduced. To understand their reductive genome evolution (RGE), we compared their genomes. They have small genomes containing chemoautotrophic and intracellular symbiotic features, and lack most genes for DNA recombination and repair e.g. recA and mutY. Their genome structures were highly conserved excepting one inversion. Many deletions from small (1 kbp up to 11 kbp) sizes were detected and deletion numbers decreased exponentially with size. Densities of deletions, short-repeat density and A+T content were higher in non-coding regions than in coding regions. Because Calyptogena symbiont genomes lack recA, we deduced that deletions and the inversion occurred by RecA-independent recombination (RIR) at short-repeats with simultaneous consumption of repeats, and that short-repeats were regenerated by high rate of mutation with enhanced A+T bias in the absence of mutY. We proposed that an active RGE is ongoing by short-repeats dependent RIR with regeneration of short-repeats in extant Calyptogena symbionts.
{"title":"The symbiotic mechanism between the chemoautotrophic symbiotic bacteria and its host deep-sea Calyptogena clams, and the reductive genome evolution of these symbionts","authors":"H. Kuwahara","doi":"10.3118/JJSE.7.2.29","DOIUrl":"https://doi.org/10.3118/JJSE.7.2.29","url":null,"abstract":"Dense animal communities at hydrothermal vents and cold seeps rely on symbioses with chemoautotrophic bacteria. To reveal the symbiotic mechanism, we sequenced the genome of chemoautotrophic intracellular symbiont in deep-sea clam, Calyptogena okutanii. On the other hand, C. magnifica symbiont genome sequenced in USA. The genomes appear to have been reduced. To understand their reductive genome evolution (RGE), we compared their genomes. They have small genomes containing chemoautotrophic and intracellular symbiotic features, and lack most genes for DNA recombination and repair e.g. recA and mutY. Their genome structures were highly conserved excepting one inversion. Many deletions from small (1 kbp up to 11 kbp) sizes were detected and deletion numbers decreased exponentially with size. Densities of deletions, short-repeat density and A+T content were higher in non-coding regions than in coding regions. Because Calyptogena symbiont genomes lack recA, we deduced that deletions and the inversion occurred by RecA-independent recombination (RIR) at short-repeats with simultaneous consumption of repeats, and that short-repeats were regenerated by high rate of mutation with enhanced A+T bias in the absence of mutY. We proposed that an active RGE is ongoing by short-repeats dependent RIR with regeneration of short-repeats in extant Calyptogena symbionts.","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"69 6 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129139524","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Microbial tetraether lipids as an environmental tracer","authors":"T. Kitajima","doi":"10.3118/JJSE.6.45","DOIUrl":"https://doi.org/10.3118/JJSE.6.45","url":null,"abstract":"","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"16 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128067655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Identification and characterization of the stator-force generator, MotPS, for motility of alkaliphilic Bacillus pseudofirumus OF4","authors":"政博 伊藤","doi":"10.3118/JJSE.3.1_42","DOIUrl":"https://doi.org/10.3118/JJSE.3.1_42","url":null,"abstract":"","PeriodicalId":204480,"journal":{"name":"Journal of Japanese Society for Extremophiles","volume":"344 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124250785","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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