Methods and Protocols最新文献

{"title":"Optimized Wound Healing Assay to Study Extracellular Vesicle-Driven Glioblastoma Cell Migration.","authors":"Concetta D'Antonio, Francesca Mantile, Gabriella Pocsfalvi, Giovanna L Liguori","doi":"10.3390/mps9030081","DOIUrl":"10.3390/mps9030081","url":null,"abstract":"<p><p>Cell migration is a fundamental process in cancer progression, playing a central role in tumor invasion and metastasis. This highly coordinated behavior is regulated by dynamic interactions between cancer cells and extracellular environment. Among the different tumor types, glioblastoma (GB) represents a particularly aggressive form of cancer in which enhanced migratory capacity is a key determinant of diffuse brain infiltration, tumor recurrence, and poor prognosis. In this context, extracellular vesicles (EVs) have emerged as important mediators, regulating cell migration in several cancer types, including GB. EVs are lipid bilayer-enclosed nano- and micro-sized particles, containing various bioactive molecules that can target specific recipient cells, thereby modulating cellular properties, including the migratory behavior. Among the available methods for studying cell migration, the wound healing assay is the most widely used. Although simple, cost-effective and not requiring sophisticated equipment, its reliability and reproducibility can be affected by technical variability and the diversity of existing protocols. Here, we present an optimized protocol for executing and analyzing a cellular wound healing assay designed to assess EV-mediated migration in GB cells. The protocol incorporates the use of silicone culture inserts to enhance wound homogeneity and reproducibility, together with continuous Mitomycin C incubation to inhibit cell proliferation without inducing cytotoxicity, enabling specific assessment of cell migration. We outline a step-by-step description of the procedure, detailing all required materials and equipment and highlighting critical steps, checkpoints, and key parameters. This method provides a robust framework for reproducible wound healing assays to investigate EV effects on GB cell migration.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"9 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13305267/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148318238","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":"Regulated Cell Death in Fungi, the Role of Metacaspases and Assay Techniques.","authors":"Derek Wilkinson","doi":"10.3390/mps9030083","DOIUrl":"10.3390/mps9030083","url":null,"abstract":"<p><p>Caspases orchestrate metazoan apoptosis, regulating processes such as embryogenesis, the death of old and infected cells and immune tolerance. Structural orthologs of caspases have been identified in bacteria, plants, protists and fungi and regulated cell death has been demonstrated in these organisms. This led some researchers to conclude that fungal metacaspases might perform a similar function to caspases. This review discusses regulated cell death, beginning with an account of RCD and the central role of caspases in mammalian RCD. It goes on to give examples of RCD in fungi, compares the structure and activity of caspase orthologs and outlines examples of metacaspase-dependent and metacaspase-independent cell death in fungi, focusing on <i>S. cerevisiae</i>. Finally, it addresses the question \"are metacaspases caspases?\", identifies alternative cell death proteases and recommends future research objectives.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"9 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13304501/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148318244","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 Simple, Rapid and Reliable Protocol for Extraction of High Quality Bacterial Genomic DNA Directly from Potato Tubers for Efficient PCR-Based Surveillance and Molecular Characterization of <i>Ralstonia solanacearum</i>.","authors":"Brian Mwangi, Joshua M Njiru, Sarah A Wandili, Kennedy K Gachoka, Kenneth Mburu, Geoffrey Muriira, Henry Rotich, Elvince Ager, Evans N Nyaboga","doi":"10.3390/mps9030084","DOIUrl":"10.3390/mps9030084","url":null,"abstract":"<p><p>Potato (<i>Solanum tuberosum</i> L.) is an important staple and food security crop to many communities in the world. However, potato production and quality is greatly constrained by bacterial wilt, a disease caused by a soil-borne pathogen, <i>Ralstonia solanacearum</i>. <i>Ralstonia solanacearum</i> can be managed through clean seed systems and therefore laboratory testing is a pre-requisite for seed certification to confirm the absence of the pathogen in potato seeds before planting. Molecular diagnostics is the gold standard for detection of <i>R. solanacearum</i> in potato seeds. However, the extraction of genomic DNA from <i>R. solanacearum</i> for molecular diagnostics is complex, tedious, lengthy and/or costly procedure. A simple, rapid and reliable DNA extraction protocol is required for use in routine molecular diagnosis of <i>R. solanacearum</i>, a high-risk quarantine pathogen. In this study, we developed a simple and rapid protocol for extracting genomic DNA from symptomatic and asymptomatic potato tubers infected with <i>R. solanacearum</i> and verified its efficiency for the detection and molecular characterization of the pathogen. The protocol was developed from the evaluation of distilled water, Tris-EDTA (TE) and Tris buffer as a base solution for tissue maceration. The DNA quantity and integrity was determined using the NanoDrop 2000C spectrophotometer and agarose gel electrophoresis, respectively. Both hot and cold solutions produced intact high molecular weight genomic DNA of sufficient yield and purity for molecular-based applications. The detection and determination of phylotypes of <i>R. solanacearum</i>, based on conventional and multiplex polymerase chain reaction (PCR), amplified the expected 280 and 372 bp amplicons, respectively, confirming that the quantity and quality of the extracted pathogen genomic DNA was sufficient for molecular diagnostic applications. The sequencing of the amplified products of the <i>endoglucanase</i> gene produced good quality sequences, which confirmed the <i>R. solanacearum</i> isolates to be members of phylotype II sequevar 1. This protocol is a simple, fast and reliable tool for the extraction of sufficient genomic DNA with high quality, directly from <i>R. solancearum</i>-infected potato tubers for PCR and sequencing applications. Its simplicity and throughput make it valuable for use in routine diagnostics and can be adopted by certification programs to ensure distribution of clean potato seeds to farmers.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"9 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13304767/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148318103","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 Simple Approach to Sample Preparation for Accurate and Reproducible Gas Phase Breakthrough Analysis of Adsorbent Materials.","authors":"Daniel A Corbin, Christopher J Breshike, Michael R Papantonakis, R Andrew McGill","doi":"10.3390/mps9030080","DOIUrl":"10.3390/mps9030080","url":null,"abstract":"<p><p>A simple approach to sample preparation for gas phase breakthrough analysis of adsorbent materials such as metal-organic frameworks is reported. To circumvent issues related to particle size, MOF powders are coated onto glass beads using only the adhesion forces between the glass surface and the particles themselves. These coatings are sufficiently stable for the coated beads to be packed into columns and used for breakthrough measurements of the pure solids. Samples prepared in this manner are compared to analogous samples coated using a binder to attach the MOF to the surface of the beads. In many cases, the approach reported here achieves higher uptake capacities and longer breakthrough times than when a binder is used, presumably because the binder partially clogs the porous structure of the MOF. In addition, an example is discussed that highlights the possibility of a reaction between the sorbent and the binder, highlighting the advantage of a simplified sample preparation method that does not require additional chemical additives.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"9 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13304718/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148318084","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":"Folate Receptor Alpha Autoantibodies in Early Pregnancy: First-Trimester Reference Intervals and Proposed Clinical Thresholds.","authors":"Claudio Giorlandino, Marina Cupellaro, Katia Margiotti, Francesca Giorlandino, Francesco Pignataro, Maria Luisa Mastrandrea, Raffaella Raffio, Laura D'Emidio, Alvaro Mesoraca, Vincenzo Milite","doi":"10.3390/mps9030079","DOIUrl":"10.3390/mps9030079","url":null,"abstract":"<p><p>Maternal folate receptor alpha autoantibodies (FRAA) have been associated with impaired placental folate transport, fetal cerebral folate deficiency (CFD) and neurodevelopmental risks including autism spectrum disorder (ASD); however, first-trimester-specific reference intervals remain undefined. This prospective single-center study of 534 healthy pregnant women at 10 + 0 to 15 + 6 weeks' gestation (week 10 n = 26; week 11 n = 155; week 12 n = 203; week 13 n = 105; week 14 n = 39; week 15 n = 6) used a CE-IVDR FRAA ELISA, following CLSI EP28-A3c and IFCC C-RIDL protocols, to establish week-specific percentiles (P5, P50, P95, P99) via non-parametric estimation and log-smoothed regression with a 1-week rolling window. An internal-consistency ROC analysis was performed against the within-dataset ≥P99 designation and is therefore not interpretable as discrimination against an independent clinical outcome. Median FRAA declined from 29 ng/mL (week 10) to 25 ng/mL (week 14), with provisional clinically actionable thresholds of P95 ≈ 120 ng/mL and P99 ≈ 150 ng/mL. These data provide the first first-trimester normative percentile curves for maternal FRAA and may support prioritizing FRAA assessment before 15 weeks (onset of accelerated transplacental IgG transfer). Given the cross-sectional, single-center design, the small samples at weeks 14-15, and the absence of long-term neurodevelopmental outcome data, the proposed thresholds and any downstream clinical implications, including folinic acid intervention and ASD risk mitigation, should be considered hypothesis-generating and require external and longitudinal validation.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"9 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13304568/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148318197","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":"The ADAPT-HEAT Study: A Multi-Method Approach to Develop Recommendations for Drug Safety During Hot Weather (The CALOR List)-Study Protocol.","authors":"Maxie Bunz, Pascal Nohl-Deryk, Heike van de Sand, Katharina van Baal, Svenja Arendt, Alina Herrmann, Ingo Meyer, Adriana Poppe, Olaf Krause, Johannes Heck, Beate Sigrid Müller","doi":"10.3390/mps9030078","DOIUrl":"10.3390/mps9030078","url":null,"abstract":"<p><p>Certain medications may adversely affect health during hot days and heatwaves by altering chronic conditions, comorbidities, fluid balance, or impairing heat adaptation. This study aims to develop evidence-based cross-sectoral recommendations for the safe administration of heat-sensitive medications, compiled into a so-called 'CALOR' list (calor: Latin for 'heat'). Development of the CALOR list will follow a four-pillar process. First, a scoping review of scientific literature and best practices will identify potentially inadequate medications during heat events (heat-PIMs) and adaptation measures, resulting in a first draft. Second, an expert panel will refine this draft through a Delphi process to reach consensus on clinically relevant recommendations. Third, German statutory health insurance (SHI) claims data will be analysed to determine heat-PIMs prevalence; data from Cologne residents will additionally be linked with climate data to investigate health outcomes during heat events. Fourth, thirty health professionals (i.e., medical doctors, nurses, pharmacists) will field-test the CALOR list in summer, providing qualitative feedback on feasibility, leading to further refinement of the CALOR list. To our knowledge this study protocol presents the first study attempting to collate a comprehensive and actionable list of recommendations for drug safety management during hot days and heatwaves.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"9 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13305179/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148318237","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 Divergent TaqMan RT-qPCR Strategy for Isoform-Resolved Detection of HIV-1 Circular RNAs.","authors":"Christopher Mauer, Sean Paz, Massimo Caputi","doi":"10.3390/mps9030077","DOIUrl":"10.3390/mps9030077","url":null,"abstract":"<p><p>The HIV-1 genome is initially transcribed as a single primary RNA that undergoes extensive splicing to produce over 40 linear and 15 circular RNA (circRNA) isoforms sharing common sequences. Conventional methods for circRNA detection, such as Northern blotting and hybridization-based assays, are inadequate for distinguishing specific circRNA isoforms when multiple circular and linear species originate from the same transcript. We previously identified 15 HIV-1 circRNAs generated by backsplicing and demonstrated that some enhance viral replication by sequestering cellular miRNAs. PCR-based approaches using divergent primers (RT-qPCR) offer greater specificity for detecting individual circular RNAs under these conditions. Building on this, we have developed a TaqMan qPCR assay capable of specifically detecting 14 HIV circRNA isoforms using backsplicing junction-directed divergent primers coupled to a hydrolysis probe for signal confirmation. Compared with matched SYBR Green assays, the TaqMan platform showed lower background in non-infected controls and reduced variance across donor-derived samples. This method provides a robust platform for selective and qualitative analysis of HIV-1 circRNAs.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"9 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13214776/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148032813","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":"Task-Specific Reliability and Measurement Error of Frontal Plane Kinematics in Individuals with Patellofemoral Pain: A Preliminary Study.","authors":"Hiraku Nagahori, Isabella Keefer, Derrick Smith, Brendan Yawn, Jing Nong Liang, Kai-Yu Ho","doi":"10.3390/mps9030076","DOIUrl":"10.3390/mps9030076","url":null,"abstract":"<p><p>This study evaluated the test-retest reliability, standard error of measurement (SEM), and minimal detectable change (MDC) of frontal plane projection angles (FPPAs) across five single-leg tasks in individuals with patellofemoral pain (PFP). Two-dimensional video data was collected from ten individuals with predominantly unilateral PFP. Participants performed single-leg squat, single-leg landing, single-leg hop, forward step-down, and lateral step-down across two testing sessions. FPPAs were measured at peak knee flexion for each task, including trunk lean angle, knee FPPA, hip FPPA, and dynamic valgus index. Test-retest reliability was assessed using intraclass correlation coefficients (ICCs). Our findings indicate that test-retest reliability and measurement error for trunk and lower limb FPPA varied across tasks in individuals with PFP. The lowest ICC was observed for hip FPPA, particularly during single-leg squat and lateral step-down tasks. Among the five tasks tested, the single-leg squat appeared to be the most demanding task, demonstrating the lowest ICCs, and highest SEM and MDC values across all four outcome measures (trunk lean angle, knee and hip FPPAs, and dynamic valgus index). The dynamic valgus index consistently showed larger SEM and MDC values than isolated hip or knee FPPAs, likely reflecting compounded measurement errors across segments. These findings provide preliminary insights, though confirmation in larger samples in persons with PFP is warranted.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"9 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13214613/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148032970","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":"Comparative Evaluation of Methods of DNA Extraction from Cryopreserved Bovine Semen for Molecular Diagnostic Applications.","authors":"Carlos A Ramos-Jonapá, Lily X Zelaya-Molina, Luis Felipe Guzmán, Edgar I González-Jiménez, David Urbán-Duarte, Horacio Álvarez-Gallardo, Francisco J Padilla-Ramírez","doi":"10.3390/mps9030075","DOIUrl":"10.3390/mps9030075","url":null,"abstract":"<p><p>Cryopreserved bovine semen represents an accessible source of genetic material due to its widespread use in assisted reproductive technologies and the conservation of genetically valuable animals. However, DNA extraction from spermatozoa within this type of sample remains challenging due to the high protein content and the complex structure of the ejaculate, which can affect DNA yield and quality. The aim of this study was to identify and validate an efficient method for obtaining high-quality DNA from spermatozoa present in cryopreserved bovine semen for molecular diagnostic applications. Five DNA extraction protocols were evaluated: TRIzol™, MagMax™ Nucleic Acid Purification Kit, Rapid DNA™ Fecal/Soil Microbe Kit, a conventional phenol-chloroform protocol, and a modified phenol-chloroform-isoamyl alcohol protocol. All extracted genetic material was assessed by spectrophotometry (concentration and purity), and DNA integrity was evaluated by agarose gel electrophoresis. Statistical analysis revealed significant differences in DNA concentration among extraction methods (Friedman test, χ² = 22.0, df = 4, <i>p</i> = 0.0002). Post hoc comparisons indicated that the modified phenol-chloroform-isoamyl alcohol protocol yielded significantly higher DNA concentrations compared to selected methods. This protocol showed the highest DNA concentration (1006.2 ± 829.4 ng/μL) and favorable purity values, and enabled consistent amplification in both conventional PCR and qPCR assays targeting the β-actin gene and <i>Tritrichomonas foetus</i>, respectively. These findings suggest that the modified protocol represents a suitable and promising approach for extracting genomic DNA from spermatozoa in cryopreserved bovine semen, with potential applications in molecular diagnostics and reproductive biotechnology.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"9 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13214769/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148032848","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":"Evaluating the Efficacy of Monoclonal Antibodies Against a Bioactive Peptide Involved in Alzheimer's Disease: A Methodological Approach.","authors":"Georgina Bonny, Kashif Mahfooz, Sara Garcia-Rates, Sibah Hasan, Susan Adele Greenfield","doi":"10.3390/mps9030074","DOIUrl":"10.3390/mps9030074","url":null,"abstract":"<p><p>Antibody treatment for Alzheimer's disease is an evolving therapeutic strategy that ensures high affinity and specificity to the target antigen; however, current approaches have proven only partially successful. A 14-mer peptide, T14, is twice as high in Alzheimer's brains and has been identified as a primary driver in the neurodegenerative process. Previously, the polyclonal antibody Ab-19 was shown to be as effective as the T14 receptor blocker (NBP-14) in reducing the toxic calcium influx in PC12 cells. The aim of this study was to establish a thorough validation process in order to evaluate the efficacy of respective anti-T14 monoclonal antibodies in T14 detection and rescuing potential from T14-induced toxicity in PC12 cells. Subsequently, we assessed the binding affinity of the most promising antibody, THK-117, via quantitative indirect conjugated T14 ELISA assays. The level of efficacy shown proved to be comparable to the polyclonal antibody, yet with the additional advantage of robust manufacturing reproducibility and high binding specificity toward the T14 epitope. With a notably low EC50, THK-117 can be viewed as a promising candidate for humanization, offering a strong potential as a therapeutic monoclonal antibody for the treatment and prevention of Alzheimer's disease.</p>","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":"9 3","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13214972/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148032854","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}