Methods and Protocols最新文献

{"title":"The Multifunctional Catalytic Hemoglobin from <i>Amphitrite ornata</i>: Protocols on Isolation, Taxonomic Identification, Protein Extraction, Purification, and Characterization.","authors":"Anna L Husted, Victoria R Sutton, Lauren A Presnar, R Kevin Blackburn, Joseph L Staton, Stephen A Borgianini, Edward L D'Antonio","doi":"10.3390/mps7060100","DOIUrl":"10.3390/mps7060100","url":null,"abstract":"<p><p>The multifunctional catalytic hemoglobin from the terebellid polychaete <i>Amphitrite ornata</i>, also named dehaloperoxidase (<i>Ao</i>DHP), utilizes the typical oxygen transport function in addition to four observed activities involved in substrate oxidation. The multifunctional ability of <i>Ao</i>DHP is presently a rare observation, and there exists a limitation for how novel dehaloperoxidases can be identified from macrobenthic infauna. In order to discover more infaunal DHP-bearing candidates, we have devised a facilitated method for an accurate taxonomic identification that places visual and molecular taxonomic approaches in parallel. Traditional visual taxonomic species identification by the non-specialist, at least for <i>A. ornata</i> or even for other marine worms, is a very difficult and time-consuming task since a large diversity is present and the method is restricted to adult worm specimens. The work herein aimed to describe a method that simplifies the taxonomic identification of <i>A. ornata</i> in particular through the assessment of its mitochondrial cytochrome c oxidase subunit I gene by employing the DNA barcoding technique. Furthermore, whole-worm specimens of <i>A. ornata</i> were used to extract and purify <i>Ao</i>DHP followed by an H₂O₂-dependent peroxidase activity assay evaluation against substrate 2,4,6-trichlorophenol. <i>Ao</i>DHP isoenzyme A was also overexpressed as the recombinant protein in <i>Escherichia coli</i>, and its peroxidase activity parameters were compared to <i>Ao</i>DHP from the natural source. The activity assay assessment indicated a tight correlation for all Michaelis-Menten parameters evaluated. We conclude that the method described herein exhibits a streamlined approach to identify the polychaete <i>A. ornata</i>, which can be adopted by the non-specialist, and the full procedure is predicted to facilitate the discovery of novel dehaloperoxidases from other marine invertebrates.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"7 6","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11678344/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142896166","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}

{"title":"Impact of the Timed and Targeted Counselling Model on Maternal Health Continuum of Care Outcomes in Northern Uganda: Protocol of a Quasi-Experimental Study.","authors":"Douglas Zibugu, Jessica S Gubbels, Christabellah Namugenyi, John Bosco Asiimwe, Sanne Gerards","doi":"10.3390/mps7060098","DOIUrl":"10.3390/mps7060098","url":null,"abstract":"<p><strong>Background: </strong>About 287,000 women died globally during their pregnancy journey in 2020, yet most of these deaths could have been prevented. In Uganda, studies show that using Community Health Worker (CHW) visits to households with a pregnant woman can support the prevention of adverse maternal and neonatal outcomes. One such intervention is through the timed and targeted counselling (ttC) approach, where CHWs deliver tailored messages to mothers and their male caregivers at key stages of pregnancy. This study aims to evaluate the impact of the ttC approach on maternal health in Northern Uganda. The main outcomes include antenatal care attendance, advised place of delivery, and postnatal care visit.</p><p><strong>Methods: </strong>We will employ a cross-sectional quasi-experimental design, with retrospective data to compare an intervention group (where ttC is implemented) to a control group (without intervention) using the propensity score matching (PSM) technique applying a 1:1 ratio with a caliper width of 20% of the standard deviation to estimate the average treatment effects. Adjusted odds ratios after generating matched pairs will be reported with 95% confidence intervals with Rosenbaum sensitivity analysis carried out for robustness.</p><p><strong>Discussion: </strong>These findings can be used to modify the implementation of the ttC approach, thereby enhancing its efficiency and effectiveness.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"7 6","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11676282/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142896145","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}

{"title":"Energy-Resolved Mass Spectrometry and Mid-Infrared Spectroscopy for Purity Assessment of a Synthetic Peptide Cyclised by Intramolecular Huisgen Click Chemistry.","authors":"Alicia Maroto, Ricard Boqué, Dany Jeanne Dit Fouque, Antony Memboeuf","doi":"10.3390/mps7060097","DOIUrl":"10.3390/mps7060097","url":null,"abstract":"<p><p>Cyclic peptides have higher stability and better properties as therapeutic agents than their linear peptide analogues. Consequently, intramolecular click chemistry is becoming an increasingly popular method for the synthesis of cyclic peptides from their isomeric linear peptides. However, assessing the purity of these cyclic peptides by mass spectrometry is a significant challenge, as the linear and cyclic peptides have identical masses. In this paper, we have evaluated the analytical capabilities of energy-resolved mass spectrometry (ER MS) and mid-infrared microscopy (IR) to address this challenge. On the one hand, mixtures of both peptides were subjected to collision-induced dissociation tandem mass spectrometry (CID MS/MS) experiments in an ion trap mass spectrometer at several excitation energies. Two different calibration models were used: a univariate model (at a single excitation voltage) and a multivariate model (using multiple excitation voltages). The multivariate model demonstrated slightly enhanced analytical performance, which can be attributed to more effective signal averaging when multiple excitation voltages are considered. On the other hand, IR microscopy was used for the quantification of the relative amount of linear peptide. This was achieved through univariate calibration, based on the absorbance of an alkyne band specific to the linear peptide, and through Partial Least Squares (PLS) multivariate calibration. The PLS calibration model demonstrated superior performance in comparison to univariate calibration, indicating that consideration of the full IR spectrum is preferable to focusing on the specific peak of the linear peptide. The advantage of IR microscopy is that it is linear across the entire working interval, from linear peptide molar ratios of 0 (equivalent to pure cyclic peptide) up to 1 (pure linear peptide). In contrast, the ER MS calibration models exhibited linearity only up to 0.3 linear peptide molar ratio. However, ER MS showed better performances in terms of the limit of detection, intermediate precision and the root-mean-square-error of calibration. Therefore, ER MS is the optimal choice for the detection and quantification of the lowest relative amounts of linear peptides.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"7 6","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11676744/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142896141","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}

{"title":"AI-Assisted High-Throughput Tissue Microarray Workflow.","authors":"Konrad Kurowski, Sylvia Timme, Melanie Christine Föll, Clara Backhaus, Philipp Anton Holzner, Bertram Bengsch, Oliver Schilling, Martin Werner, Peter Bronsert","doi":"10.3390/mps7060096","DOIUrl":"10.3390/mps7060096","url":null,"abstract":"<p><p>Immunohistochemical (IHC) studies of formalin-fixed paraffin-embedded (FFPE) samples are a gold standard in oncology for tumor characterization, and the identification of prognostic and predictive markers. However, despite the abundance of archived FFPE samples, their research use is limited due to the labor-intensive nature of IHC on large cohorts. This study aimed to create a high-throughput workflow using modern technologies to facilitate IHC biomarker studies on large patient groups. Semiautomatic constructed tissue microarrays (TMAs) were created for two tumor patient cohorts and IHC stained for seven antibodies (ABs). AB expression in the tumor and surrounding stroma was quantified using the AI-supported image analysis software QuPath. The data were correlated with clinicopathological information using an R-script, all results were automatically compiled into formatted reports. By minimizing labor time to 7.7%-compared to whole-slide studies-the established workflow significantly reduced human and material resource consumption. It successfully correlated AB expression with overall patient survival and additional clinicopathological data, providing publication-ready figures and tables. The AI-assisted high-throughput TMA workflow, validated on two patient cohorts, streamlines modern histopathological research by offering cost and time efficiency compared to traditional whole-slide studies. It maintains research quality and preserves patient tissue while significantly reducing material and human resources, making it ideal for high-throughput research centers and collaborations.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"7 6","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11678066/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142896140","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}

{"title":"A Critical Exploration of the Total Flavonoid Content Assay for Honey.","authors":"Sharmin Sultana, Ivan Lozada Lawag, Lee Yong Lim, Kevin J Foster, Cornelia Locher","doi":"10.3390/mps7060095","DOIUrl":"10.3390/mps7060095","url":null,"abstract":"<p><p>This study critically investigates the aluminium chloride-based colorimetric determination of the total flavonoid content (TFC) of honey. Following a comprehensive review of the recent literature reporting the use of the assay in the determination of TFC in honey, 10 honeys of different botanical origins were investigated using the colorimetric method alongside an artificial honey that was used as a control. Using spiking experiments, this study demonstrates that the flavonoid concentrations commonly found in honey are too low for a direct measurement and thus some of the TFC data reported in the literature might more likely be a reflection of the honey's inherent colour rather than a product of the coordination complex formed specifically between flavonoids and Al³⁺ ions. This paper highlights the importance of correct blanking and suggests alternative approaches to the traditional TFC assay for honey to ensure analysis results that are truly reflective of honey's TFC.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"7 6","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11586951/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710658","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}

{"title":"Digital PCR Validation for Characterization of Quantitative Reference Material of <i>Escherichia coli</i> O157<i>:H7</i> Genomic DNA.","authors":"Claudia Patricia Tere-Peña, Martha Nancy Calderon-Ozuna, John Emerson Leguizamón Guerrero","doi":"10.3390/mps7060094","DOIUrl":"10.3390/mps7060094","url":null,"abstract":"<p><p><i>Escherichia coli O157:H7</i>, a Shiga-toxin-producing <i>E. coli</i> (STEC), is an important pathogen related to foodborne disease that is responsible for a growing number of outbreaks worldwide and has been detected in processed meats, dairy, and fresh vegetables. Although culturing is the gold standard method for detection of this bacterium, molecular methods based on nucleic acid amplification techniques such as PCR are becoming more common because of their rapidity, sensitivity, and specificity. However, to ensure reliable results among the several alternative PCR protocols (e.g., commercial kits and reference methods), different measurement assurance tools, including validated methods, reference materials, and proficiency tests, among others, are required. Herein, we present a digital PCR method validation for <i>E. coli O157:H7</i> detection and quantification using seven specific gene sequences; this method quantified nucleic acids from different <i>E. coli</i> serotypes, with a detection range of 6.6 to 7900 copies/µL and a repeatability standard deviation over the concentration range of 1% to 13.6%. The relative standard uncertainty was 3.5-14.6%, and the detection limit was 0.27 copies/µL. Subsequently, two batches of a candidate reference material based on <i>E. coli</i> O157:H7 genomic DNA were then produced and characterized for evaluation of copy number concentration with the validated ddPCR method, with assigned values of 164,770 ± 9251 and 172 ± 9 copies/μL. Thus, this study demonstrated the development of a validated method and reference material for dPCR and qPCR detection of <i>E. coli</i> O157:H7, a key STEC responsible for food poisoning.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"7 6","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11587158/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710763","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}

{"title":"A Reproducible Protocol for the Isolation of Malaria-Derived Extracellular Vesicles by Differential Centrifugation.","authors":"Tosin Opadokun, Petra Rohrbach","doi":"10.3390/mps7060092","DOIUrl":"10.3390/mps7060092","url":null,"abstract":"<p><p>Over the last few decades, malaria-derived extracellular vesicles (EVs) have gained increasing interest due to their role in disease pathophysiology and parasite biology. Unlike other EV research fields, the isolation of malaria EVs is not standardized, hampering inter-study comparisons. Most malaria EV studies isolate vesicles by the \"gold-standard\" technique of differential (ultra)centrifugation (DC). Here, we describe in detail an optimized and reproducible protocol for the isolation of malaria-derived EVs by DC. The protocol begins with a description of cultivating high-parasitemia, synchronous <i>P. falciparum</i> cultures that are the source of EV-containing conditioned culture media. The isolation protocol generates two EV subtypes, and we provide details of characterizing these distinct subtypes by analyzing human and parasite proteins by Western blot analysis. We identify some of these proteins as suitable markers for malaria EV subpopulations and subtypes.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"7 6","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11587005/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710662","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}

{"title":"Identifying and Removing Fraudulent Attempts to Enroll in a Human Health Improvement Intervention Trial in Rural Communities.","authors":"Karla L Hanson, Grace A Marshall, Meredith L Graham, Deyaun L Villarreal, Leah C Volpe, Rebecca A Seguin-Fowler","doi":"10.3390/mps7060093","DOIUrl":"10.3390/mps7060093","url":null,"abstract":"<p><p>Using the internet to recruit participants into research trials is effective but can attract high numbers of fraudulent attempts, particularly via social media. We drew upon the previous literature to rigorously identify and remove fraudulent attempts when recruiting rural residents into a community-based health improvement intervention trial. Our objectives herein were to describe our dynamic process for identifying fraudulent attempts, quantify the fraudulent attempts identified by each action, and make recommendations for minimizing fraudulent responses. The analysis was descriptive. Validation methods occurred in four phases: (1) recruitment and screening for eligibility and validation; (2) investigative periods requiring greater scrutiny; (3) baseline data cleaning; and (4) validation during the first annual follow-up survey. A total of 19,665 attempts to enroll were recorded, 74.4% of which were considered fraudulent. Automated checks for IP addresses outside study areas (22.1%) and reCAPTCHA screening (10.1%) efficiently identified many fraudulent attempts. Active investigative procedures identified the most fraudulent cases (33.7%) but required time-consuming interaction between researchers and individuals attempting to enroll. Some automated validation was overly zealous: 32.1% of all consented individuals who provided an invalid birthdate at follow-up were actively contacted by researchers and could verify or correct their birthdate. We anticipate fraudulent responses will grow increasingly nuanced and adaptive given recent advances in generative artificial intelligence. Researchers will need to balance automated and active validation techniques adapted to the topic of interest, population being recruited, and acceptable participant burden.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"7 6","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11587125/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710773","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}

{"title":"Near-Infrared Spectroscopy for Growth Estimation of <i>Spirulina platensis</i> Cultures.","authors":"Lamprini Malletzidou, Eleni Kyratzopoulou, Nikoletta Kyzaki, Evangelos Nerantzis, Nikolaos A Kazakis","doi":"10.3390/mps7060091","DOIUrl":"10.3390/mps7060091","url":null,"abstract":"<p><p>The present study proposes the use of Near-Infrared (NIR) spectroscopy as a rapid method for estimating the growth of <i>Spirulina platensis</i> cultures, avoiding any sample manipulation or pretreatment. NIR spectroscopy in diffuse reflectance mode was used on culture volumes as received, with Principal Component Analysis (PCA) and Partial Least Squares (PLS) linear regression, for developing the calibration model in the wavelength range of 1000-2500 nm, in order to choose the appropriate wavelength to estimate the growth of the microalga. The local reflectance maximum at 1062.6 nm, connected with reduced water absorption and scattering effects by the microalga, was identified from PCA as the positive peak in the first loading plot, correlating diffuse reflectance with dilution levels. The calibration curve of diffuse reflectance at 1062.6 nm in response to dilution presented strong linearity, supported by a coefficient of determination (R²) of 0.995. Cross-validation of NIR spectra with a <i>S. platensis</i> culture confirmed the method's reliability, showing that the growth follows an exponential pattern. The study shows that diffuse reflectance NIR spectroscopy can be used for the rapid monitoring of <i>Spirulina platensis</i> growth.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"7 6","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11586962/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710794","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}

{"title":"Optimizing Arterial Tissue Thickness Measurement Protocols: Digital Vernier Caliper Versus Digital Thickness Gauge.","authors":"Alexandru Petru Ion, Alexandra Asztalos, Claudiu Constantin Ciucanu, Eliza Russu, Adrian Vasile Mureșan, Eliza-Mihaela Arbănași, Traian V Chirilă, Gabriela Strnad, Emil-Marian Arbănași","doi":"10.3390/mps7060090","DOIUrl":"10.3390/mps7060090","url":null,"abstract":"<p><strong>Background: </strong>The aim of this study is to analyze the reproducibility of sample thickness measurements taken by a non-experienced user by comparing a standard digital vernier caliper, with four different protocols, to a specialized thickness gauge.</p><p><strong>Methods: </strong>The current study is a methodological study where we examined the thickness of the porcine arterial wall in the thoracic aorta of six pigs. Two adjacent samples of 10 × 10 mm from each aorta were excised longitudinally from the anterior wall, resulting in twelve specimens. Five protocols were employed to measure the thickness of each sample. In four of these protocols, digital vernier calipers (Multicomp PRO MP012475) were utilized, while the fifth protocol utilized a specialized digital thickness gauge (Mitutoyo 547-500S, Mitutoyo Corp., Kawasaki, Japan).</p><p><strong>Results: </strong>We observed a higher average thickness of the samples during the initial measurement compared to the second measurement (1.11 ± 0.16 vs. 0.94 ± 0.17, <i>p</i> = 0.0319) with the first protocol and smaller values than those determined at the last measurement (0.93 ± 0.15 vs. 1.10 ± 0.15, <i>p</i> = 0.0135) for the third protocol. Further, with the digital vernier calipers, we recorded lower values for all four protocols than for the digital thickness gauge determinations. In addition, we computed the ratio of the thicknesses measured during the first, second, and third measurements to analyze how consistent the values were across the three consecutive measurements, with no difference regarding the third, fourth, and control protocols.</p><p><strong>Conclusions: </strong>The digital thickness gauge offers dependable measurements, regardless of the user's expertise in assessing tissue thickness, and demonstrates a substantially higher reproducibility when compared to the digital vernier. We also found that taking an average of the thickness measurements from four specific points on each half of the sides or on each diagonal of each corner yielded consistently reliable results over time when using a standard digital vernier caliper instead of a specialized one.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"7 6","pages":""},"PeriodicalIF":2.3,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11587071/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710813","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}