Microbial Biotechnology最新文献

{"title":"Genetic Bioaugmentation-Mediated Bioremediation of Terephthalate in Soil Microcosms Using an Engineered Environmental Plasmid.","authors":"Alejandro Marquiegui-Alvaro, Anastasia Kottara, Micaela Chacón, Lisa Cliffe, Michael Brockhurst, Neil Dixon","doi":"10.1111/1751-7915.70071","DOIUrl":"10.1111/1751-7915.70071","url":null,"abstract":"<p><p>Harnessing in situ microbial communities to clean-up polluted natural environments is a potentially efficient means of bioremediation, but often the necessary genes to breakdown pollutants are missing. Genetic bioaugmentation, whereby the required genes are delivered to resident bacteria via horizontal gene transfer, offers a promising solution to this problem. Here, we engineered a conjugative plasmid previously isolated from soil, pQBR57, to carry a synthetic set of genes allowing bacteria to consume terephthalate, a chemical component of plastics commonly released during their manufacture and breakdown. Our engineered plasmid caused a low fitness cost and was stably maintained in terephthalate-contaminated soil by the bacterium P. putida. Plasmid carriers efficiently bioremediated contaminated soil in model soil microcosms, achieving complete breakdown of 3.2 mg/g of terephthalate within 8 days. The engineered plasmid horizontally transferred the synthetic operon to P. fluorescens in situ, and the resulting transconjugants degraded 10 mM terephthalate during a 180-h incubation. Our findings show that environmental plasmids carrying synthetic catabolic operons can be useful tools for in situ engineering of microbial communities to perform clean-up even of complex environments like soil.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 1","pages":"e70071"},"PeriodicalIF":5.7,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11725763/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142968805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Oral Immunisation With Non-GMO Surface Displayed SARS-CoV-2 Spike Epitopes on Bacteria-Like Particles Provokes Robust Humoral and Cellular Immune Responses, and Modulated the Gut Microbiome in Mice.","authors":"Robie Vasquez, Ji Hoon Song, Remilyn M Mendoza, In-Chan Hwang, Bernadette B Bagon, Lars Engstrand, Valerie Diane Valeriano, Dae-Kyung Kang","doi":"10.1111/1751-7915.70073","DOIUrl":"10.1111/1751-7915.70073","url":null,"abstract":"<p><p>The coronavirus disease 2019 (COVID-19) is a fatal disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). To date, several vaccines have been developed to combat the spread of this virus. Mucosal vaccines using food-grade bacteria, such as Lactobacillus spp., are promising strategies for developing safe and effective vaccines against SARS-CoV-2. In this study, we designed a non-GMO surface-displayed SARS-CoV-2 spike S1 epitope on Limosilactobacillus fermentum-derived bacteria-like particles (BLPs). After that, we evaluated its efficacy to induce immune responses in immunocompetent mice. Moreover, we examined the influence of oral immunisation on the gut microbiome and microbiota metabolites. Twenty-eight 6-week-old male C57BL/6 mice were orally immunised with the following: PBS (control), Lm. fermentum-derived BLPs only, BLPs displaying SARS-CoV-2 spike S1-2, or BLPs displaying SARS-CoV-2 spike S1-3 epitopes. Our results showed that mucosal immunisation of mice with surface-displayed SARS-CoV-2 spike epitopes provoked high-level secretory IgA and systemic IgG production. Moreover, the immunisation exhibited a Th1-like immune response, characterised by an elevated IgG2a-to-IgG1 ratio and high antiviral IFN-γ production. In addition, we observed gut microbiome modulation and increased butyrate production in immunised mice. Overall, the use of Lm. fermentum-derived BLPs and the anchor CshA to display SARS-CoV-2 spike S1epitopes is a promising novel strategy in developing a cost-effective, non-GMO mucosal vaccine alternative against SARS-CoV-2.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 1","pages":"e70073"},"PeriodicalIF":5.7,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11724470/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142963244","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microbes Saving Lives and Reducing Suffering.","authors":"Kenneth Timmis, Zeynep Ceren Karahan, Juan Luis Ramos, Omry Koren, Ana Elena Pérez-Cobas, Karen Steward, Victor de Lorenzo, Elisabetta Caselli, Margaret Douglas, Clarissa Schwab, Virginia Rivero, Rafael Giraldo, Junkal Garmendia, Raymond J Turner, Jessamyn Perlmutter, José M Borrero de Acuña, Pablo Ivan Nikel, Jerome Bonnet, Angela Sessitsch, James K Timmis, Carla Pruzzo, M Auxiliadora Prieto, Siavash Isazadeh, Wei E Huang, Gerard Clarke, Danilo Ercolini, Max Häggblom","doi":"10.1111/1751-7915.70068","DOIUrl":"https://doi.org/10.1111/1751-7915.70068","url":null,"abstract":"","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 1","pages":"e70068"},"PeriodicalIF":5.7,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143021449","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Easy-Curing and pH-Regulated CRISPR-Cas9 Plasmids for Gene Editing and Plasmid Curing in Lactococcus cremoris","authors":"Javier Nicolás Garay-Novillo,&nbsp;José Ángel Ruiz-Masó,&nbsp;Gloria del Solar,&nbsp;José Luis Barra","doi":"10.1111/1751-7915.70060","DOIUrl":"https://doi.org/10.1111/1751-7915.70060","url":null,"abstract":"<p>In this work, we developed a plasmid-based CRISPR-Cas9 strategy for editing <i>Lactococcus cremoris</i>, which allows easy generation of plasmid-free strains with the desired modification. We constructed versatile shuttle vectors based on the theta-type pAMβ1 promiscuous replicon and p15A <i>ori</i>, expressing both the Cas9 nuclease gene (under pH-regulated promoters derived from P170) and a single-guide RNA for specific targeting (under a strong constitutive promoter). The vectors designed for plasmid targeting were very effective for low- and high-copy-number plasmid curing in <i>L. cremoris</i>, and their targeting efficiency was shown to be tunable by regulating <i>cas9</i> expression. For chromosome editing, we implemented a host-independent method that enhances double-homologous recombination events using plasmids expressing the genes encoding λRed-phage Redβ recombinase and <i>Escherichia coli</i> single-stranded DNA binding protein (EcSSB). By coupling either the endogenous recombination machinery or the Redβ-EcSSB-assisted recombination system with our novel chromosome-targeting CRISPR-Cas9 plasmids, we efficiently generated and selected thousands of gene-edited cells. Examination of the impact of the constructed CRISPR-Cas9 vectors on host fitness revealed no Cas9-associated toxicity, and, remarkably, these vectors exhibited a very high loss rate when growing the bacterial host cells in the absence of selective pressure.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 12","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70060","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142868877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Easy-Curing and pH-Regulated CRISPR-Cas9 Plasmids for Gene Editing and Plasmid Curing in Lactococcus cremoris","authors":"Javier Nicolás Garay-Novillo,&nbsp;José Ángel Ruiz-Masó,&nbsp;Gloria del Solar,&nbsp;José Luis Barra","doi":"10.1111/1751-7915.70060","DOIUrl":"https://doi.org/10.1111/1751-7915.70060","url":null,"abstract":"<p>In this work, we developed a plasmid-based CRISPR-Cas9 strategy for editing <i>Lactococcus cremoris</i>, which allows easy generation of plasmid-free strains with the desired modification. We constructed versatile shuttle vectors based on the theta-type pAMβ1 promiscuous replicon and p15A <i>ori</i>, expressing both the Cas9 nuclease gene (under pH-regulated promoters derived from P170) and a single-guide RNA for specific targeting (under a strong constitutive promoter). The vectors designed for plasmid targeting were very effective for low- and high-copy-number plasmid curing in <i>L. cremoris</i>, and their targeting efficiency was shown to be tunable by regulating <i>cas9</i> expression. For chromosome editing, we implemented a host-independent method that enhances double-homologous recombination events using plasmids expressing the genes encoding λRed-phage Redβ recombinase and <i>Escherichia coli</i> single-stranded DNA binding protein (EcSSB). By coupling either the endogenous recombination machinery or the Redβ-EcSSB-assisted recombination system with our novel chromosome-targeting CRISPR-Cas9 plasmids, we efficiently generated and selected thousands of gene-edited cells. Examination of the impact of the constructed CRISPR-Cas9 vectors on host fitness revealed no Cas9-associated toxicity, and, remarkably, these vectors exhibited a very high loss rate when growing the bacterial host cells in the absence of selective pressure.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 12","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70060","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142868929","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Penitentiaries: Bringing microbiological literacy to the fringes of society","authors":"Víctor de Lorenzo","doi":"10.1111/1751-7915.70052","DOIUrl":"10.1111/1751-7915.70052","url":null,"abstract":"<p>This report highlights a science outreach effort for prisons launched by the Spanish National Research Council (CSIC) in collaboration with the NGO Solidarios para el Desarrollo. The Microbiology-focused part of the initiative aims at educating inmates on some basic facts, in order to raise awareness about microorganisms and their impact on daily life. The outline of the talks, inspired by the International Initiative for Microbial Literacy, aims to encourage this collective to move from passive listeners into active participants, helping them understand that Earth is a microbial planet, and that their bodies harbour vast microbiomes that affect their health and social interactions. The talks introduce Microbiology using simple metaphors and emphasize the role of beneficial microorganisms. By explaining the power of microscopes, inmates are shown the hidden microbial world that surrounds them, sparking interest and curiosity. The talks also cover microbial biotechnology, using examples such as bioplastics, anti-cavity bacteria, and skin microorganisms designed for acne prevention. Overall, this outreach initiative seeks to provide inmates with valuable scientific knowledge, fostering curiosity and critical thinking. Despite the challenges of delivering such content in a prison setting, the initiative demonstrates that even marginalized groups can benefit from microbiological literacy, helping them to both endure their terms and eventually reintegrate into society.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 12","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70052","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142862709","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Desert-adapted plant growth-promoting pseudomonads modulate plant auxin homeostasis and mitigate salinity stress","authors":"Ramona Marasco,&nbsp;Maria J. Mosqueira,&nbsp;Kholoud A. Seferji,&nbsp;Sarah M. Al Romaih,&nbsp;Grégoire Michoud,&nbsp;Jian Xu,&nbsp;Cristina Bez,&nbsp;Tatiana Castillo Hernandez,&nbsp;Vittorio Venturi,&nbsp;Ikram Blilou,&nbsp;Daniele Daffonchio","doi":"10.1111/1751-7915.70043","DOIUrl":"10.1111/1751-7915.70043","url":null,"abstract":"<p>By providing adaptive advantages to plants, desert microorganisms are emerging as promising solutions to mitigate the negative and abrupt effects of climate change in agriculture. Among these, pseudomonads, commonly found in soil and in association with plants' root system, have been shown to enhance plant tolerance to salinity and drought, primarily affecting root system architecture in various hosts. However, a comprehensive understanding of how these bacteria affect plant responses at the cellular, physiological and molecular levels is still lacking. In this study, we investigated the effects of two <i>Pseudomonas</i> spp. strains, E102 and E141, which were previously isolated from date palm roots and have demonstrated efficacy in promoting drought tolerance in their hosts. These strains colonize plant roots, influencing root architecture by inhibiting primary root growth while promoting root hair elongation and lateral root formation. Strains E102 and E141 increased auxin levels in <i>Arabidopsis</i>, whereas this effect was diminished in IAA-defective mutant strains, which exhibited reduced IAA production. In all cases, the effectiveness of the bacteria relies on the functioning of the plant auxin response and transport machinery. Notably, such physiological and morphological changes provide an adaptive advantage to the plant, specifically under stress conditions such as salinity. Collectively, this study demonstrates that by leveraging the host's auxin signalling machinery, strains E102 and E141 significantly improve plant resilience to abiotic stresses, positioning them as potential biopromoters/bioprotectors for crop production and ecosystem restoration in alignment with Nature-based Solution approaches.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 12","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70043","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142845477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploration of In Situ Extraction for Enhanced Triterpenoid Production by Saccharomyces cerevisiae","authors":"Mariam Dianat,&nbsp;Sarah Straaten,&nbsp;Aldo Maritato,&nbsp;Daniel Wibberg,&nbsp;Tobias Busche,&nbsp;Lars M. Blank,&nbsp;Birgitta E. Ebert","doi":"10.1111/1751-7915.70061","DOIUrl":"10.1111/1751-7915.70061","url":null,"abstract":"<p>Plant-derived triterpenoids are in high demand due to their valuable applications in cosmetic, nutraceutical, and pharmaceutical industries. To meet this demand, microbial production of triterpenoids is being developed for large-scale production. However, a prominent limitation of microbial synthesis is the intracellular accumulation, requiring cell disruption during downstream processing. Destroying the whole-cell catalyst drives up production costs and limits productivity and product yield per cell. Here, in situ product extraction of triterpenoids into a second organic phase was researched to address this limitation. An organic solvent screening identified water-immiscible isopropyl myristate as a suitable in situ extractant, enabling extraction of up to 90% of total triterpenoids from engineered <i>Saccharomyces cerevisiae</i>. Combining isopropyl myristate and β-cyclodextrins improved extraction efficiency. In a first configuration, repeated batch fermentation with sequential product extraction and cell recycling resulted in 1.8 times higher production than a reference fermentation without in situ product extraction. In the second configuration, yeast cells were in contact with the second organic phase throughout a fed-batch fermentation to continuously extract triterpenoids. This resulted in 90% product extraction and an extended production phase. Further improvement of triterpenoid production was not achieved due to microbial host limitations uncovered through omics analyses.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 12","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70061","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142851624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Actinomycetota From Macroalgae as Rich Source for Natural Products Discovery Revealed Through Culture-Dependent and -Independent Approaches","authors":"Mariana Girão,&nbsp;Adriana Rego,&nbsp;Ana C. Fonseca,&nbsp;Weiwei Cao,&nbsp;Zhongjun Jia,&nbsp;Ralph Urbatzka,&nbsp;Pedro N. Leão,&nbsp;Maria F. Carvalho","doi":"10.1111/1751-7915.70058","DOIUrl":"10.1111/1751-7915.70058","url":null,"abstract":"<p>Actinomycetota are unrivalled producers of bioactive natural products, with strains living in association with macroalgae representing a prolific—yet largely unexplored—source of specialised chemicals. In this work, we have investigated the bioactive potential of Actinomycetota from macroalgae through culture-dependent and -independent approaches. A bioprospecting pipeline was applied to a collection of 380 actinobacterial strains, recovered from two macroalgae species collected in the Portuguese northern shore—<i>Codium tomentosum</i> and <i>Chondrus crispus—</i>in order to explore their ability to produce antibacterial, antifungal, anticancer and lipid-reducing compounds. Around 43% of the crude extracts showed activity in at least one of the screenings performed: 111 presented antimicrobial activity at 1 mg/mL, 83 significantly decreased cancer cells viability at 15 μg/mL and 5 reduced lipid content in zebrafish &gt; 60% at 15 ug/mL. Dereplication of active extracts unveiled the presence of compounds that could explain most of the recorded results, but also unknown molecules in the metabolome of several strains, highlighting the opportunity for discovery. The bioactive potential of the actinobacterial community from the same macroalgae specimens, which served as the source for the aforementioned Actinomycetota collection, was also explored through metagenomics analysis, allowing to obtain a broader picture of its functional diversity and novelty. A total of 133 biosynthetic gene clusters recovered from metagenomic contigs and metagenome assembled genomes (MAGs). These were grouped into 91 gene cluster families, 83 of which shared less than 30% of similarity to database entries. Our findings provided by culture-dependent and -independent approaches underscore the potential held by actinomycetes from macroalgae as reservoirs for novel bioactive natural products.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 12","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70058","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142845475","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Heterologous Gene Expression in Chlamydomonas reinhardtii Chloroplast by Heterologous Promoters and Terminators, Intercistronic Expression Elements and Minichromosome","authors":"Yunling Guo,&nbsp;Hui Xiong,&nbsp;Qiuling Fan,&nbsp;Deqiang Duanmu","doi":"10.1111/1751-7915.70069","DOIUrl":"10.1111/1751-7915.70069","url":null,"abstract":"<p><i>Chlamydomonas reinhardtii</i>, a model green alga for expressing foreign proteins, faces challenges in multigene expression and enhancing protein expression level in the chloroplast. To address these challenges, we compared heterologous promoters, terminators and intercistronic expression elements (IEEs). We transformed Chlamydomonas chloroplast with a biolistic approach to introduce vectors containing the NanoLuc expression unit regulated by Chlamydomonas or tobacco promoters and terminators. We observed that tobacco promoters P<i>rbcL</i> and P<i>psbA</i> could not effectively regulate protein expression, whereas tobacco terminators T<i>rbcL</i> and T<i>rps16</i> did not affect the expression of Nluc protein. Further exploration of IEEs specific to Chlamydomonas revealed that Cr-IEE2 had a minor effect on both upstream and downstream protein expression, whereas Cr-IEE5 significantly influenced downstream protein expression. In contrast, tobacco IEE was found to be unsuitable for driving protein expression in Chlamydomonas. Additionally, VOR element and Rep protein derived from beet curly top geminivirus were able to form a minichromosome in Chlamydomonas chloroplast, and this system could enhance protein expression level compared to the traditional method of site-specific integration in the plastome. This study highlights the potential of IEEs and minichromosome in facilitating heterologous protein expression in Chlamydomonas chloroplast.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"17 12","pages":""},"PeriodicalIF":5.7,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70069","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142833328","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}