Engineering Microbiology最新文献

{"title":"Decoupling growth phase dependency and metal ion inhibition: A dual engineering strategy for the high-yield biosynthesis of microcin J25 in Escherichia coli","authors":"Guangxin Yang ,&nbsp;Xinchan Wang ,&nbsp;Yunting Zhou ,&nbsp;Xiuliang Ding ,&nbsp;Jinxiu Huang ,&nbsp;Shiyan Qiao ,&nbsp;Aihua Deng ,&nbsp;Haitao Yu","doi":"10.1016/j.engmic.2025.100230","DOIUrl":"10.1016/j.engmic.2025.100230","url":null,"abstract":"<div><div>Microcin J25 (MccJ25) has received substantial attention as a potential solution to the global threat of infection caused by antibiotic-resistant bacteria. However, the industrial fermentation of MccJ25 faces production bottlenecks. It is imperative to further explore the production optimization strategies for MccJ25 to formulate comprehensive approaches for its industrial-scale production and other downstream applications. Here, Fe²⁺ in tap water was identified as a critical inhibitor of MccJ25 biosynthesis, selectively repressing <em>mcjA</em> transcription, which was reversible via 2,2′-bipyridine-mediated chelation. To decouple production from growth phase dependency and Fe²⁺ interference, we engineered <em>Escherichia coli</em> BL21 cells by performing two genetic modifications. First, we replaced the native <em>mcjA</em> promoter with a constitutive promoter (P_Q) to allow its mid-log phase expression. Second, we replaced the native <em>mcjBCD</em> promoter with a medium-strength variant (P₂₂₂₃) that delayed production kinetics without affecting final yields. However, the genomic integration of <em>mcjD</em> alleviated plasmid-borne toxicity, increasing the expression timing and doubling the yield to 240 mg/L. Finally, we computationally optimized the <em>mcjA</em> ribosome-binding site (RBS) to enhance translation efficiency. RBS optimization revealed that a moderate translation initiation efficiency (550,584 arbitrary units [au]) maximized production, whereas excessive efficiency (2,019,712 au) impaired growth and output. These interventions synergistically increased the MccJ25 titer 10-fold, reaching 430 mg/L in batch culture. Our findings establish a robust platform for MccJ25 overproduction, highlighting promoter engineering and translational tuning as pivotal strategies for antimicrobial peptide biosynthesis. This study provides insights for overcoming metabolic constraints in microbial fermentation, advancing the development of peptide-based therapeutics against multidrug-resistant pathogens.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 4","pages":"Article 100230"},"PeriodicalIF":0.0,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145121026","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":"Unraveling microbial life from a high-altitude hydrothermal system in the Andes plateau and their potential for nitrogen transformations","authors":"Coral Pardo-Esté ,&nbsp;Juan Castro-Severyn ,&nbsp;Jacqueline Aldridge ,&nbsp;Diego Alvarez-Saravia ,&nbsp;Lenka Kurte ,&nbsp;Polette Aguilar-Muñoz ,&nbsp;Pablo Paquis ,&nbsp;Vilma Pérez ,&nbsp;David Medina ,&nbsp;Wade H Jeffrey ,&nbsp;Verónica Molina ,&nbsp;Martha B Hengst","doi":"10.1016/j.engmic.2025.100242","DOIUrl":"10.1016/j.engmic.2025.100242","url":null,"abstract":"<div><div>Terrestrial hydrothermal systems provide a window for studying the biogeochemical interactions that occur in hot and gas-rich ecosystems resembling the conditions found in early life on Earth. The biogeochemical dynamics of the Andean hydrothermal systems in the Atacama Desert area are still understudied. Thus, we aimed to characterize the taxonomic composition and genomic potential of nitrogen transformations in a microbial community inhabiting a high-altitude hydrothermal system on the Altiplano Plateau of the Chilean Andes. Specifically, we sampled sediment and microbial mats in three ponds with water temperatures ranging from 42 to 64 °C. We found a high prevalence of photoheterotrophs, with differences in taxonomic composition and gene abundance between the microbial communities found in the sediment and microbial mats. Changes in physicochemical conditions, such as temperature and pH, and the concentrations of CO₂, CH₄ and Mn accounted for the variability in the microbial community structure. Our results indicated an enrichment of N-related genes associated with nitrate reduction, denitrification, and ammonia assimilation, suggesting a metabolically versatile community using nitrate, nitrite, and gaseous nitrogen species to assimilate ammonia into their biomass. This study contributes to our understanding of the taxonomy and functional microbial dynamics in a high-altitude thermal system, where ammonia assimilation is potentially critical for biomass formation, and particular environmental conditions favor adaptations to maintain biogeochemical cycles.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 4","pages":"Article 100242"},"PeriodicalIF":0.0,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145519646","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":"The role of microbiota dysbiosis in Parkinson’s disease: Pathophysiology and therapeutic opportunities","authors":"Shabnam Santos ,&nbsp;Ivonne Salinas ,&nbsp;Nicolás Almeida ,&nbsp;Andrés Caicedo","doi":"10.1016/j.engmic.2025.100222","DOIUrl":"10.1016/j.engmic.2025.100222","url":null,"abstract":"<div><div>Parkinson’s disease (PD) is a chronic, progressive neurodegenerative disorder characterized by debilitating motor and non-motor symptoms. Its etiology is multifactorial, with no single definitive cause identified, although aging is a significant risk factor. Additional risks include genetic predisposition, family history, and environmental factors such as pesticide exposure and <em>Helicobacter pylori</em> infection. Dysbiosis of the gut microbiota, and in particular bacterial imbalances, has been implicated in the disruption of the gut-brain axis, contributing to both systemic and neuroinflammation. Environmental factors such as antibiotic exposure and toxins can precipitate microbial dysregulation, potentially accelerating PD progression. Understanding the mechanisms of the gut-brain axis and identifying strategies to preserve a healthy microbiome are essential for developing novel therapeutic approaches. This review synthesizes current therapeutic strategies and ongoing research focused on restoring gut-brain balance to combat PD. These approaches include fecal microbiota transplantation, dietary interventions, and probiotic therapies, all of which show promise in mitigating both motor and non-motor symptoms. Furthermore, we emphasize the urgent need for continued research into probiotics and innovative therapeutic approaches for gut-brain axis modulation, presenting novel opportunities for effective PD management.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 3","pages":"Article 100222"},"PeriodicalIF":0.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144703538","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":"Production of dicarboxylates from ω-amino acids using a cofactor- and co-substrate-free in vitro biosynthetic system","authors":"Jinxin Yan ,&nbsp;Hui Zhang ,&nbsp;Hongxu Zhang ,&nbsp;Hairong Yu ,&nbsp;Wenjia Tian ,&nbsp;Mingyuan Liu ,&nbsp;Weikang Sun ,&nbsp;Leilei Guo ,&nbsp;Xiaoxu Tan ,&nbsp;Kaiyu Gao ,&nbsp;Tianyi Jiang ,&nbsp;Chuanjuan Lü ,&nbsp;Qianjin Kang ,&nbsp;Wensi Meng ,&nbsp;Cuiqing Ma ,&nbsp;Chao Gao ,&nbsp;Ping Xu","doi":"10.1016/j.engmic.2025.100210","DOIUrl":"10.1016/j.engmic.2025.100210","url":null,"abstract":"<div><div>Dicarboxylates are valuable platform compounds with a broad range of applications. The <em>in vitro</em> biosynthetic system used to produce dicarboxylates from ω-amino acids via the natural pathway requires costly cofactors and co-substrates, which restricts its economic feasibility of use. In this study, we designed a cofactor- and co-substrate-free artificial pathway for the production of dicarboxylates from ω-amino acids. Only three enzymes (viz., amine oxidase from <em>Kluyveromyces marxianus</em> DMKU3-1042, xanthine oxidase from bovine milk, and catalase from <em>Aspergillus niger</em>) were required for dicarboxylate production. Succinate (0.79 g g^-1), glutarate (0.83 g g^-1), and adipate (0.77 g g^-1) were produced in high yields from the corresponding ω-amino acids through the <em>in vitro</em> biosynthetic system with the artificial pathway. Glutarate could also be produced from <span>l</span>-lysine by further introducing <span>l</span>-lysine monooxygenase and 5-aminovaleramide amidohydrolase from <em>Pseudomonas putida</em> KT2440 into the <em>in vitro</em> biosynthetic system, with the cofactor- and co-substrate-free system achieving a product yield of 0.63 g g^-1. Considering its desirable characteristics, this artificial pathway-based <em>in vitro</em> biosynthetic system may be a promising alternative for dicarboxylate production from biotechnologically produced ω-amino acids.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 3","pages":"Article 100210"},"PeriodicalIF":0.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144272099","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":"Genotypic and molecular characterization of a moderately thermophilic cyanobacterium, Gloeocapsa sp. strain BRSZ","authors":"Sasiprapa Samsri ,&nbsp;Tanwalee Deprom ,&nbsp;Chananwat Kortheerakul ,&nbsp;Sophon Sirisattha ,&nbsp;Stephen B. Pointing ,&nbsp;Hakuto Kageyama ,&nbsp;Rungaroon Waditee-Sirisattha","doi":"10.1016/j.engmic.2025.100226","DOIUrl":"10.1016/j.engmic.2025.100226","url":null,"abstract":"<div><div>A unicellular-colonial cyanobacterium, designated “BRSZ,” was isolated from a neutral-alkaline hot spring in Thailand. Morphological characterization revealed distinctive features consistent with those of the genus <em>Gloeocapsa</em>. Physiological assessments demonstrated that BRSZ is a moderately thermophilic and halotolerant cyanobacterium with the potential for chemoheterotrophic growth in dark conditions. Molecular phylogenetic analysis based on 16S ribosomal RNA (rRNA) gene sequences placed BRSZ within a well-defined <em>Gloeocapsa</em> clade, a finding corroborated by 16S–23S internal transcribed spacer (ITS) rRNA secondary structure analyses. Genome comparisons, including average nucleotide identity (ANI), genome-to-genome distance (GGD), and digital DNA-DNA hybridization (dDDH), between strain BRSZ and closely related taxa showed an ANI value of 95.45 %, near the lower boundary of the species delineation threshold (95–96 %). A GGD of 0.0374 (&gt;0.0258) and dDDH of 69 % (&lt;70 %) further supported genomic differentiation. Genome-based analysis revealed a mycosporine-like amino acid biosynthetic gene cluster likely involved in sunscreen compound production. Cultivation-based production of a UV-absorbing compound confirmed the functional relevance of this gene cluster. These findings expand the described diversity within the <em>Gloeocapsa</em> complex and enhance our understanding of the taxonomy of this group. In addition, they underscored the importance of hot spring environments as sources of novel extremophiles.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 3","pages":"Article 100226"},"PeriodicalIF":0.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144866338","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":"A novel therapeutic strategy of methicillin-resistant Staphylococcus aureus","authors":"Ying Wang ,&nbsp;Mengyan Xu ,&nbsp;Hanne Ingmer","doi":"10.1016/j.engmic.2025.100231","DOIUrl":"10.1016/j.engmic.2025.100231","url":null,"abstract":"<div><div><em>Staphylococcus aureus</em> is a major public health threat, largely due to its remarkable capacity to develop antimicrobial resistance. Zhang <em>et al.</em> recently demonstrated a highly innovative approach to eradicate chronic methicillin-resistant <em>S. aureus</em> infections by inducing bacterial calcification with antibody-polysialic acid conjugates targeting wall teichoic acids, while simultaneously modulating host immune responses via enhanced calprotectin expression and macrophage activation. Despite limitations, this strategy represents a promising and unconventional therapy to combat resistant <em>S. aureus</em> infections.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 3","pages":"Article 100231"},"PeriodicalIF":0.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144885416","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":"Screening of molecular elements and improvement of heat resistance in a thermophilic bacterium","authors":"Jie Cui ,&nbsp;Caifeng Li ,&nbsp;Gongze Cao ,&nbsp;Yuxia Wu ,&nbsp;Shouying Xu ,&nbsp;Youming Zhang ,&nbsp;Xiaoying Bian ,&nbsp;Qiang Tu ,&nbsp;Wentao Zheng","doi":"10.1016/j.engmic.2025.100225","DOIUrl":"10.1016/j.engmic.2025.100225","url":null,"abstract":"<div><div>Engineering microorganisms to withstand extreme temperatures (&gt;80 °C) remains a critical challenge in industrial biotechnology owing to limited genetic tools and poor mechanistic understanding of microbial thermoadaptation. We aimed to develop a novel <em>Geobacillus stearothermophilus</em> strain with remarkable thermal resilience through an integrated approach combining adaptive laboratory evolution and rational genetic engineering. Progressive thermal adaptation (70–80 °C) followed by genome reduction generated a mutant (SL-1–80) with enhanced stability at 80 °C. Subsequent combinatorial overexpression of eight heat-associated genes (<em>murD, cysM, grpE, groES, hsp33, hslO, hrcA, clpE</em>) synergistically extended its survival to 85 °C. Genomic and transcriptomic analyses revealed a triple mechanism: (1) strategic deletion of transposable elements (IS5377/IS4/IS110) reduced genomic instability, (2) co-activation of chaperone systems (GroES-GrpE) and redox homeostasis enzymes (HslO<img>Hsp33) enhanced protein folding and oxidative stress resistance, and (3) metabolic plasticity (BglG and HTH-domain transcriptional repressor), motility optimization (FliY), and transcriptional reprogramming (Sigma-D, DUF47-family chaperone and HTH-domain transcriptional repressor) facilitated nutrient acquisition and motility-based environmental navigation under stress. Furthermore, we established the first high-efficiency electroporation protocol (10⁴ transformants/µg DNA) for this genus, enabling ATP-enhanced heterologous protein expression under heat stress. This study provided a robust platform organism for high-temperature bioprocessing and a mechanistic blueprint for engineering microbial thermotolerance, addressing key limitations in applications such as microbial-enhanced oil recovery and industrial enzyme production.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 3","pages":"Article 100225"},"PeriodicalIF":0.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144866337","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":"Analysis of single-cell RNA sequencing data to examine the gastric inflammation-to-cancer transition and evaluation of the effect of probiotic on precancerous lesions","authors":"Minmin Hu ,&nbsp;Shiyang Xu ,&nbsp;Ruofei Xu ,&nbsp;Xiangjie Qi ,&nbsp;Xiaofeng Yu ,&nbsp;Jinqi Wang ,&nbsp;Yige Li ,&nbsp;Yangyang Liu ,&nbsp;Guiran Xi ,&nbsp;Junbao Yu ,&nbsp;Mei Shi","doi":"10.1016/j.engmic.2025.100208","DOIUrl":"10.1016/j.engmic.2025.100208","url":null,"abstract":"<div><div>Gastric cancer (GC) is the fifth most prevalent malignancy globally. However, its heterogeneity and asymptomatic early-stage development hinder timely diagnosis and effective treatment. Here, we employed single-cell RNA sequencing to delineate the transitional features of pit mucous cells (PMCs) during the gastritis-to-cancer transition and identified 100 core genes. Characterization of the gene set revealed the role of ribosomal protein small subunit and ribosomal protein large subunit in inflammation-to-cancer transition, which promoted ribonucleoprotein complex biogenesis and cytoplasmic translation. External validation using independent cohorts confirmed that this core gene set discriminated disease progression (AUC &gt; 0.7) and was significantly enriched in GC tissues (<em>p</em> &lt; 0.01). Moreover, we evaluated the therapeutic intervention effects of <em>C. butyricum</em> and synbiotics (Weichanghao®) using a rat model of gastritis and demonstrated the targeted suppression of inflammation-to-cancer transition genes. Our findings establish the basis for early diagnosis of GC through PMC-driven molecular dynamics. Additionally, we propose microbiota-based strategies to prevent the inflammation-to-cancer transition in preneoplastic stages. Furthermore, our results highlight that dysbiosis of the gastric microbiome can be addressed using probiotic supplementations and the core gene set may provide labeling for the evaluation of probiotics-based treatment.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 3","pages":"Article 100208"},"PeriodicalIF":0.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144205430","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":"Production of cycloastragenol in metabolically engineered yeast","authors":"Jingxian Zhang ,&nbsp;Peng Xu ,&nbsp;Yongjun Wei","doi":"10.1016/j.engmic.2025.100227","DOIUrl":"10.1016/j.engmic.2025.100227","url":null,"abstract":"<div><div>Cycloastragenol is a bioactive, high-value triterpenoid derived from <em>Astragalus membranaceus</em>. Conventional plant-based extraction and chemical synthesis methods are expensive. To our knowledge, this is the first report on the <em>de novo</em> biosynthesis of cycloastragenol in yeast. The mevalonate pathway was reconstituted in yeast peroxisomes, and the engineered yeast produced 656.55 mg/L squalene. Further introduction of heterologous enzymes led the engineered yeast to produce 1.04 mg/L cycloastragenol, which demonstrated the yeast production of value-added medicinal molecules.</div></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 3","pages":"Article 100227"},"PeriodicalIF":0.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144831441","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":"Tumor-targeting bioluminescent bacteria for <i>in vivo</i> imaging.","authors":"Chenghao Ma, Jingxi Liu, Hongjing Liu, Xiaohan Zhao, Geng Li, Youming Zhang, Tianyu Jiang","doi":"10.1016/j.engmic.2025.100224","DOIUrl":"https://doi.org/10.1016/j.engmic.2025.100224","url":null,"abstract":"<p><p>As the understanding of bacteria-mediated cancer therapies deepens, bacteria such as <i>Escherichia coli</i> Nissle 1917 (EcN) have become a promising platform for cancer therapy. However, their potential role in real-time monitoring and visualization tools still needs to be explored and enhanced. In this study, we aimed to screen and optimize EcN visualization systems for non-invasive <i>in vivo</i> bioluminescence imaging in live mice. To this end, we developed three series of recombinant EcN strains expressing <i>Gaussia</i> luciferase (Gluc), <i>Renilla</i> luciferase (Rluc), and NanoLuc (Nluc), along with their respective mutants. These strains exhibited bioluminescence when different coelenterazine (CTZ) substrates were present. As a result, multiple bioluminescent EcN strain-substrate pairs were identified with stronger, longer, or red-shifted bioluminescence, offering multiple effective optical tumor-targeting systems for <i>in vivo</i> studies investigating bacteria-mediated cancer therapy and intestinal diseases.</p>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"5 3","pages":"100224"},"PeriodicalIF":0.0,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12967832/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147694558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}