Biosensors and Bioelectronics: X最新文献

{"title":"Fluorescent probes to track complex membrane blebbing","authors":"Juan L. Cortes-Muñoz ,&nbsp;Johan-Moritz Kux ,&nbsp;Pablo J. Sáez ,&nbsp;Arturo Jiménez-Sánchez","doi":"10.1016/j.biosx.2025.100584","DOIUrl":"10.1016/j.biosx.2025.100584","url":null,"abstract":"<div><div>Cellular blebbing, pivotal in processes such as apoptosis, cytokinesis, and migration, involves dynamic interactions between the actomyosin network and microtubules. However, existing probes inadequately capture these simultaneous interactions, limiting our ability to study blebbing mechanisms. We present <strong>AztecBleb</strong> probes (<strong>AztecBleb</strong>^{<strong>DAPI</strong>}, <strong>AztecBleb</strong>^{<strong>GFP</strong>}, <strong>AztecBleb</strong>^{<strong>TxR</strong>}, <strong>AztecBleb</strong>^{<strong>Cy5</strong>}), novel fluorescent reporters designed to selectively target and monitor blebbing in real-time. These probes incorporate a pregnenolone-based scaffold as a hydrophobic core derived from abiraterone acetate, facilitating precise localization to blebs and microtubules without disrupting cellular function. Through persistent staining of cell blebs, these photostable and biocompatible probes enable continuous monitoring of blebbing and microtubule dynamics during cellular migration. Our approach provides new insights into the coordination of bleb formation and cytoskeletal remodeling, offering a unique tool for studying motility-driven cellular behaviors.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100584"},"PeriodicalIF":10.61,"publicationDate":"2025-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143161324","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":"Enhanced glucose sensing via nanoparticles-modified extended gates: A novel approach to electric double layer modulation and signal amplification in field effect transistors for improved detection sensitivity","authors":"Sheng-Chun Hung,&nbsp;Yi-Hua Lee","doi":"10.1016/j.biosx.2025.100576","DOIUrl":"10.1016/j.biosx.2025.100576","url":null,"abstract":"<div><div>This study presents an innovative glucose sensing platform that harnesses the enhanced electrocatalytic properties of planar electric double layer (EDL) structures in conjunction with extended gate field effect transistors (EGFETs). By integrating specific nanoparticles onto the sensor surface, this platform achieves substantial improvements in glucose detection sensitivity and overall performance. The materials employed in this research include Ni nanowires combined with graphene films, CuO nanoparticles incorporated into carbon nanostructures, and gold nanoparticles affixed to ZnO nanostructures. These nanomaterials exhibit remarkable catalytic activity, while the localized electric field effect generated by the electric double layer significantly amplifies the signal, thereby enhancing sensitivity. Experimental findings reveal notable enhancements in both sensitivity and detection limits compared to conventional glucose sensors, underscoring the potential of this platform for effective glucose monitoring. Specifically, the Ni nanowire-graphene film sensor recorded a sensitivity of 3102.7 μA mM⁻¹ cm⁻² with a detection limit of 51 nM. The CuO nanoparticle-carbon nanostructure sensor achieved a sensitivity of 2206.25 μA mM⁻¹ cm⁻² and a detection limit of 39 nM, while the gold nanoparticle-ZnO nanostructure sensor demonstrated a sensitivity of 811.8 μA mM⁻¹ cm⁻² with a detection limit of 59 nM.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100576"},"PeriodicalIF":10.61,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143161323","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 comprehensive review of Interferometric Reflectance Imaging Sensor as a sensitive detection platform and its application areas","authors":"Monireh Bakhshpour-Yucel ,&nbsp;Nese Lortlar Unlu ,&nbsp;Elif Seymour ,&nbsp;Adil Denizli","doi":"10.1016/j.biosx.2025.100574","DOIUrl":"10.1016/j.biosx.2025.100574","url":null,"abstract":"<div><div>The Interferometric Reflectance Imaging Sensor (IRIS) technology represents a significant advancement in biosensing, providing a label-free, selective, sensitive, and high-throughput platform for detecting molecular interactions. This review explores the underlying principles, instrumentation, and diverse applications of IRIS, with a focus on its efficacy for real-time monitoring of DNA-protein and protein-protein interactions, as well as virus detection. IRIS can measure DNA hybridization kinetics and identify pathogens without labeling, highlighting its versatility and reliability in biomedical research and diagnostics. IRIS achieves enhanced sensitivity and specificity by leveraging spectral reflectivity as a transduction mechanism and employing a 3D polymeric surface chemistry for bioreceptor immobilization. The review underscores IRIS's potential to revolutionize clinical diagnostics, biomolecular screening, and the study of biomolecular binding affinities, establishing it as a powerful tool for future research and medical applications.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"22 ","pages":"Article 100574"},"PeriodicalIF":10.61,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143159522","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":"Personal glucose meter-based portable method for creatine kinase assay","authors":"Jinhwan Lee ,&nbsp;Yejin Oh ,&nbsp;Hyoyong Kim,&nbsp;Sangmo Lee,&nbsp;Hyun Gyu Park","doi":"10.1016/j.biosx.2025.100575","DOIUrl":"10.1016/j.biosx.2025.100575","url":null,"abstract":"<div><div>We herein developed a simple and label-free method to detect the creatine kinase (CK) activity by employing a personal glucose meter (PGM) as a detection device. This method relies on the target-induced consumption of glucose by the cascade enzymatic reaction (CER) promoted by the combined activities of CK and hexokinase (HK). The target CK first catalyzes the phosphorylation of adenosine 5′-diphosphate (ADP) to adenosine 5′-triphosphate (ATP) using creatine phosphate (CP) as a phosphate donor. HK then phosphorylates glucose into glucose-6-phosphate by converting the produced ATP back to ADP, which could enter another cycle of the CER to further phosphorylate glucose in a sample. As a consequence, the glucose level would decrease depending on the CK activity, which could be very conveniently detected using a PGM. With this unique design principle, we successfully determined the CK activity down to 0.0147 U/mL with high specificity against various non-target enzymes. We also verified that the developed strategy can reliably detect CK activity in heterogeneous human blood, ensuring its robust utility in various clinical applications. This user-friendly method enables point-of care detection of CK, offering a practical tool for monitoring muscle health. By allowing simultaneous tracking of CK and glucose levels with the same PGM device, this approach is particularly beneficial for diabetes mellitus (DM) patients, empowering them to proactively manage their health and reduce the risk of muscle-related complications.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"22 ","pages":"Article 100575"},"PeriodicalIF":10.61,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143159658","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":"Fiber-tip photonic crystal for real-time referenced biosensing in serum","authors":"Mathias Dolci ,&nbsp;Paco Dreverman ,&nbsp;Mildred S. Cano-Velázquez ,&nbsp;Arthur L. Hendriks ,&nbsp;Emiel Veth ,&nbsp;P.J. van Veldhoven ,&nbsp;Andrea Fiore ,&nbsp;Peter Zijlstra","doi":"10.1016/j.biosx.2024.100573","DOIUrl":"10.1016/j.biosx.2024.100573","url":null,"abstract":"<div><div>Fiber optic sensors have become increasingly well-established due to the many advantages they provide such as immunity to electromagnetic interference, multiplexing capabilities, and remote sensing. The coupling of light with a transducer at the tip of the optical fiber enables the detection of physical and biological parameters. 2D photonic crystals (PhC) can be designed to feature guided-mode resonances (GMR) characterized by a strong electric field at the PhC surface, providing a suitable tool for the detection of local refractive index variations (e.g. biomolecule adsorption). Here, we demonstrate the use of a PhC transferred to the tip of a single-mode fiber for biosensing. The control of surface chemistry provides a sensitive platform for the molecular recognition of antibody biomarkers. By integrating the fiber in a continuous flow platform, the real-time detection of anti-IgG in undiluted serum was achieved, with a limit of detection down to 60 pM. Moreover, the use of a reference channel is demonstrated to correct for signal drifts in real-time due to changes in bulk refractive index. These referenced fiber-tip PhC biosensors may pave the way for fluidic integrated systems in environmental, industrial, and healthcare applications, and open up the possibility of biosensing in the human body by integrating them into catheters.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"22 ","pages":"Article 100573"},"PeriodicalIF":10.61,"publicationDate":"2024-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143159659","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":"Development of vitex-derived polymer nanofibers using electrochemical sensors for the treatment of polycystic ovarian syndrome in rats as an animal model","authors":"Zaid H. Mahmoud ,&nbsp;Uday Abdul-Reda Hussein ,&nbsp;Najwa Aljbory ,&nbsp;Mohammed Jawad Alnajar ,&nbsp;Laleh Maleknia ,&nbsp;Abolfazl Mirani ,&nbsp;Ehsan kianfar","doi":"10.1016/j.biosx.2024.100570","DOIUrl":"10.1016/j.biosx.2024.100570","url":null,"abstract":"<div><h3>Background and objectives</h3><div>Nanofibers have a high specific surface area and small pores, which increases the possibility of drug uptake. In this study, chasteberry-containing nanofibers were prepared by electrospinning to investigate their potential for polycystic follicles after induction of polycystic ovarian syndrome (PCOS).</div></div><div><h3>Materials and methods</h3><div>Chasteberry-containing nanofibers were identified using Fourier-transform infrared spectroscopy (FTIR), Field Emission Scanning Electron Microscopy (FESEM), and thermal gravimetric analysis (TGA) tests, as well as drug release and antibacterial tests. In this study, 42 Sprague-Dawley rats with regular menstrual cycles were divided into five groups: control group (healthy rats), polycarbonate (PC) group (treated with 1 mg/kg letrozole for 28 days to induce PCOS syndrome), and Poly terephthalate (PT) group (treated with PCOS for 30 days). Chasteberry was included at 1, 2, 4 wt%. To induce PCOS, the polyterephthalate (PT) and polycarbonate (PC) groups were administered 1 mg/kg letrozole by oral gavage for 28 days. The polyterephthalate (PT) group was treated with different doses of chasteberry-containing nanofibers (1, 2, 4 wt%).</div></div><div><h3>Results</h3><div>The results showed that the drug was well incorporated into the fibers and released slowly and sustainedly from the nanofibers for 30 days. Ex vivo results showed that by taking letrozole, the polycarbonate (PC) group had larger cystic follicles with a very thin granulosa layer than the control group. This is the cause of ovarian anovulation or oligo-ovulation and induction of polycystic ovarian syndrome. In the polyterephthalate (PT) group, which was administered nanofibers containing chasteberry nanomedicine subcutaneously, the number of cystic follicles decreased and the number of various follicles indicating ovulation increased in these groups.</div></div><div><h3>Conclusion</h3><div>Subcutaneous administration of Vitex nanofibers at a dose of 4 wt% can improve polycystic ovarian syndrome by decreasing cystic follicles and increasing the number of various follicles and corpora lutea.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"22 ","pages":"Article 100570"},"PeriodicalIF":10.61,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143159519","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":"Simultaneous detection of human neutrophil elastase and cathepsin G on a single substrate using a fluorometric quantum dots probe and chemometric models","authors":"Fátima A.R. Mota,&nbsp;Rafael C. Castro,&nbsp;David S.M. Ribeiro,&nbsp;João L.M. Santos,&nbsp;Ricardo N.M.J. Páscoa,&nbsp;Marieta L.C. Passos,&nbsp;M. Lúcia M.F.S. Saraiva","doi":"10.1016/j.biosx.2024.100571","DOIUrl":"10.1016/j.biosx.2024.100571","url":null,"abstract":"<div><div>Human neutrophil elastase (HNE) and cathepsin G (CatG) are crucial proteolytic enzymes involved in the pathophysiology of chronic wounds. High levels of these enzymes indicate prolonged inflammation and impaired healing processes, making their discrimination and quantification essential for effective wound management and treatment strategies. In this study, we propose a novel method combining distinctly sized CdTe quantum dots (QDs) as a fluorescent probe to implement a platform for simultaneous discrimination and quantification of HNE and CatG, applying chemometric analysis.</div><div>The fluorometric response was acquired using two different methods: kinetic and excitation/emission matrices (EEM). These second-order data were processed using various chemometric models, including unfolded partial least-squares with residual bilinearization (U-PLS/RBL), radial basis function artificial neural network (RBF-ANN), and partial least squares-discriminant analysis (PLS-DA), to guarantee a detailed and precise analysis. The results showed that the kinetic method, when processed with the aforementioned models, accurately quantified CatG in the presence of HNE with a REP of around 20%. This method also successfully discriminated the two enzymes both together and individually, achieving a sensitivity and specificity of 1. In contrast, the EEM method only allowed for the discrimination of the two enzymes both together and individually.</div><div>Our groundbreaking approach proved to be accurate for both the discrimination and quantification of one of the enzymes, offering the advantage of being simpler and faster than other reference procedures. This method paves the way for more effective therapeutic interventions and could initiate a path toward the simultaneous discrimination of multiple enzymes.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"22 ","pages":"Article 100571"},"PeriodicalIF":10.61,"publicationDate":"2024-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143159520","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 Biologically Inspired and Protein-Based Bio-Cyber Interface for the Internet of Bio-Nano Things","authors":"Pit Hofmann ,&nbsp;Juan A. Cabrera ,&nbsp;Gunnar Schulte ,&nbsp;Frank H.P. Fitzek","doi":"10.1016/j.biosx.2024.100572","DOIUrl":"10.1016/j.biosx.2024.100572","url":null,"abstract":"<div><div>The Internet of Things (IoT) has changed the way how we interact with our physical environment, enabling connectivity and communication between physical and virtual entities, e.g., for digital twin applications. However, as we step beyond the IoT, developing the Internet of Bio-Nano Things (IoBNT), where biological and nanoscale entities will be included in our communication networks, includes challenges as well as opportunities. In the IoBNT, the role of Bio-Cyber Interfaces (BCIs) is still underscored, representing necessary building blocks that ensure the bidirectional information exchange between biological and digital communication systems. The application area of the IoBNT spans diverse domains, e.g., healthcare, personalized medicine, or environmental monitoring. This work proposes a theoretical framework for a BCI, leveraging advances in biotechnology, nanotechnology, and communication engineering to establish an interface for exchanging information between biological entities, nanoscale devices, and the digital world. Therefore, we discuss the key components of the proposed BCI framework. Furthermore, we survey the existing literature of biologically inspired BCIs and outline potential use cases and benefits of integrating BCIs in the IoBNT for various domains, such as healthcare and environmental monitoring.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"22 ","pages":"Article 100572"},"PeriodicalIF":10.61,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143159660","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":"Practical considerations for DNA sensing using Faradaic electrochemical impedance spectroscopy on ENIG PCB electrodes","authors":"Shruti Ahuja ,&nbsp;Avani Kulkarni ,&nbsp;Richa Pandey ,&nbsp;Kiran Kondabagil ,&nbsp;Siddharth Tallur","doi":"10.1016/j.biosx.2024.100569","DOIUrl":"10.1016/j.biosx.2024.100569","url":null,"abstract":"<div><div>Electroless nickel immersion gold (ENIG) finish printed circuit board (PCB) substrates offer a cost-effective solution for electrochemical sensing of biomolecules. Electrochemical impedance spectroscopy (EIS) is highly sensitive and can differentiate between bioelectrochemical circuit elements over a wide frequency range. Traditional Faradaic EIS measurements often focus on charge transfer resistance <span><math><mrow><mo>(</mo><msub><mrow><mi>R</mi></mrow><mrow><mi>c</mi><mi>t</mi></mrow></msub><mo>)</mo></mrow></math></span> to determine analyte concentration. However, the long measurement time required to perform such EIS measurements down to very low frequencies (typically 0.1<!--> <!-->Hz) can adversely affect the ENIG PCB due to gold layer degradation and copper corrosion. To overcome these issues, we propose using the constant phase element (CPE) in the electrical equivalent circuit in EIS measurements for DNA sensing. Our approach employs ENIG PCB electrodes functionalized with thiolated single-stranded DNA probes targeting the uidA gene of <em>E. coli</em>. We evaluate the specificity of this sensing scheme to a 166<!--> <!-->bp complementary amplicon from <em>E. coli</em> against non-complementary amplicons of different lengths from <em>E. coli</em> and bacteriophage Phi6. By operating at higher frequencies (<span><math><mo>&gt;</mo></math></span>10<!--> <!-->Hz), CPE analysis reduces measurement times and minimizes the risk of PCB degradation. Preliminary findings indicate that the CPE impedance exhibits concentration-dependent changes with increasing target DNA concentrations. We also present preliminary results for utilizing this sensing mechanism to detect <em>E. coli</em> from wastewater samples.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"22 ","pages":"Article 100569"},"PeriodicalIF":10.61,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143159524","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":"Electrochemical sensors for the detection of immune checkpoint related proteins and their role in cancer companion diagnostics","authors":"Louise Barnaby ,&nbsp;Andrew G. Watts ,&nbsp;Pedro Estrela","doi":"10.1016/j.biosx.2024.100561","DOIUrl":"10.1016/j.biosx.2024.100561","url":null,"abstract":"<div><div>Cancer companion diagnostics are incredibly important in helping to determine whether a patient will benefit from immune checkpoint inhibitor (ICI) treatment. Determining the chances of treatment success helps to inform clinicians to make the best treatment decisions for a particular patient. Many immune checkpoint related proteins show potential as biomarkers for ICI success, such as the checkpoint proteins themselves, cytokines, interleukins and other immune response related proteins. The most investigated checkpoint inhibitor protein is Programmed Death Ligand 1 (PD-L1), which is used as a biomarker in clinical diagnostic tests but, with some limitations. In the near future, tests for many different biomarkers will start becoming commercially available along with tests for multiple biomarkers simultaneously, giving an even better prediction of potential ICI success. Electrochemical sensors are a high sensitivity point of care diagnostic technique that can have the potential to achieve detection of multiple biomarkers at once. The main problem facing this field is improving their sensitivity to be able to detect the incredibly low concentrations of biomarkers found in liquid biopsy samples. Many methods such as enhancing an electrode surface with high conductivity materials or increasing the measured electrochemical signal via signal amplifying molecules have been investigated with promising results. This review investigates the potential biomarkers relevant to predicting ICI success, as well as the current electrochemical sensors that have been developed to determine the expression levels of these proteins.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"22 ","pages":"Article 100561"},"PeriodicalIF":10.61,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143159521","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}