Nanotheranostics最新文献

{"title":"Metal-Organic Frameworks for Bioimaging: Strategies and Challenges.","authors":"Yanfei Liu, Ting Jiang, Zhenbao Liu","doi":"10.7150/ntno.63458","DOIUrl":"10.7150/ntno.63458","url":null,"abstract":"<p><p>Metal-organic frameworks (MOFs), composited with metal ions and organic linkers, have become promising candidates in the biomedical field own to their unique properties, such as high surface area, pore-volume, tunable pore size, and versatile functionalities. In this review, we introduce and summarize the synthesis and characterization methods of MOFs, and their bioimaging applications, including optical bioimaging, magnetic resonance imaging (MRI), computed tomography (CT), and multi-mode. Furthermore, their bioimaging strategies, remaining challenges and future directions are discussed and proposed. This review provides valuable references for the designing of molecular bioimaging probes based on MOFs.</p>","PeriodicalId":36934,"journal":{"name":"Nanotheranostics","volume":" ","pages":"143-160"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8671950/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39656488","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":"Pharmacokinetic/Pharmacodynamic Determinations of Iron-tannic Molecular Nanoparticles with its Implication in MR Imaging and Enhancement of Liver Clearance.","authors":"Thipjutha Phatruengdet, Piyachat Khuemjun, Jannarong Intakhad, Saowalak Krunchanuchat, Arpamas Chariyakornkul, Rawiwan Wongpoomchai, Chalermchai Pilapong","doi":"10.7150/ntno.63310","DOIUrl":"10.7150/ntno.63310","url":null,"abstract":"<p><p>Assessment and enhancement of liver clearance are promising strategies for protection of liver from various liver diseases. Iron-tannic nanoparticles (FTs) were previously considered as imageable autophagic enhancers with biodegradation potential. Herein, we present a new approach for utilizing Iron-tannic nanoparticles (FTs) as a tool for imaging and increasing liver clearance. Pharmacokinetic profiling suggested that FTs were initially found in blood circulation and thereafter were distributed to the liver. By using MR imaging (T₁ weighted), maximum MRI signal enhancement was found to occur after 30 minutes post-injection (i.v.) and gradually decreased afterward. Decreasing MRI signal may be due to FTs metabolism by the liver. By assessing imaging-derived pharmacokinetics, we can simply determine the rate constant of liver degradation of FTs. Potentially, we might use this parameter to monitor liver function, where its clearance is of concern. Once functional implication of FTs in liver clearance was investigated, FTs were found to induce hepatocyte autophagy along with activation of lysosomes. Consequently, the hepatocytes were capable of efficiently clearing cellular debris. From these results, it is clear that FTs should be considered as a molecular tool for quantitative MRI-derived liver function assessment, and for enhancing clearance function in liver parenchyma. Hopefully, our findings will pave the way to develop new strategies for non-invasive assessment and enhancement of liver clearance.</p>","PeriodicalId":36934,"journal":{"name":"Nanotheranostics","volume":" ","pages":"195-204"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8671955/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39892501","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 CD44E/s dual-targeting DNA aptamer as nanoprobe to deliver treatment in hepatocellular carcinoma.","authors":"Cario Wing-Sze Lo,&nbsp;Cecilia Ka Wing Chan,&nbsp;Jianqing Yu,&nbsp;Mian He,&nbsp;Chung Hang Jonathan Choi,&nbsp;James Yun Wong Lau,&nbsp;Nathalie Wong","doi":"10.7150/ntno.62639","DOIUrl":"https://doi.org/10.7150/ntno.62639","url":null,"abstract":"<p><p><b>Background:</b> Hepatocellular carcinoma (HCC) is the predominant subtype of liver cancer with an extraordinary high mortality. Resistance to systemic therapy is a major cause of inferior clinical outcome in most patients with HCC. CD44 is a transmembrane cell-surface glycoprotein that is characterized by its variants displaying differential overexpression in human cancers. Aptamers, also known as chemical antibodies, can target cell-surface molecules with high affinity and specificity via structural recognition. Aptamer-mediated drug delivery hence is of high potentials in guiding therapy to improve efficacy. <b>Methods:</b> Variants CD44E and CD44s were studied for HCC relevance by investigating their expressions in primary HCC tumors, adjacent cirrhotic/fibrotic livers and normal livers using junction specific primers in qPCR assay. CD44E/s dual-targeted aptamers were uncovered by integrating loss-gain cell-SELEX and next generation sequencing. Selected aptamers were characterized for binding affinity and specificity, biostability, <i>in vivo</i> and <i>in vitro</i> cytotoxicity, <i>in vivo</i> homing and biodistribution, and ability to deliver 5-FU into targeted cells <i>in vitro</i>. <b>Results:</b> Both CD44E and CD44s isoforms showed significant upregulations in HCC tumors with CD44E/s activities promoting cell proliferation and migration. Loss-gain cell-SELEX uncover a CD44E/s dual-targeting aptamer, termed CD44-Apt1. Strong binding of CD44-Apt1 to cell-surface CD44 positive cells but not CD44-negative cells was demonstrated by flow-cytometry. CD44-Apt1 displayed strong affinity to CD44E and CD44s with K_D as low as 1 nM but not the hyaluronic acid binding domain of CD44. Confocal imaging of CD44-positive cells stained with fluorescent-labeled CD44-Apt1 showed profound cytoplasmic localization, suggesting efficient cell-penetrating ability. Meanwhile, no apparent staining was observed in CD44-negative cells. CD44-Apt1 when conjugated with inhibitor 5-FU showed efficient guidance of 5-FU into HCC cells that significantly enhanced drug toxicity by more than thousands-fold. Both <i>in vitro</i> cell treatment and <i>in vivo</i> animal biodistribution indicated that CD44-Apt1 is non-toxic. In HCC xenograft model, CD44-Apt1 efficiently homed to tumor xenografts in a CD44 expression-dependent manner. <b>Conclusion:</b> Novel discovery of aptamer CD44-Apt1 that can bind both CD44E and CD44s illustrates high potential as nanoprobe to deliver anti-cancer therapeutics.</p>","PeriodicalId":36934,"journal":{"name":"Nanotheranostics","volume":" ","pages":"161-174"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8671951/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39656489","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":"Ultrasound-Guided Microbubble-Mediated Locoregional Delivery of Multiple MicroRNAs Improves Chemotherapy in Hepatocellular Carcinoma.","authors":"Huaijun Wang,&nbsp;Zhongqian Hu,&nbsp;Uday Kumar Sukumar,&nbsp;Rajendran Jc Bose,&nbsp;Arsenii Telichko,&nbsp;Jeremy J Dahl,&nbsp;Ramasamy Paulmurugan","doi":"10.7150/ntno.63320","DOIUrl":"10.7150/ntno.63320","url":null,"abstract":"<p><p><b>Rationale:</b> To assess treatment effects of 4 complementary miRNAs (miRNA-100/miRNA-122/antimiRNA-10b/antimiRNA-21) encapsulated in a biodegradable PLGA-PEG nanoparticle, administered by an ultrasound-guided microbubble-mediated targeted delivery (UGMMTD) approach in mouse models of hepatocellular carcinoma (HCC). <b>Methods:</b><i>In vitro</i> apoptotic index was measured in HepG2 and Hepa1-6 HCC cells treated with various combinations of the 4 miRNAs with doxorubicin. Three promising combinations were further tested <i>in vivo</i> by using UGMMTD. 63 HepG2 xenografts in mice were randomized into: group 1, miRNA-122/antimiRNA-10b/antimiRNA-21/US/doxorubicin; group 2, miRNA-100/miRNA-122/antimiRNA-10b/antimiRNA-21/US/doxorubicin; group 3, miRNA-100/miRNA-122/antimiRNA-10b/US/doxorubicin; group 4, miRNA-122/anitmiRNA-10b/antimiRNA-21/doxorubicin; group 5, miRNA-100/miRNA-122/antimiRNA-10b/antimiRNA-21/doxorubicin; group 6, miRNA-100/miRNA-122/antimiRNA-10b/doxorubicin; group 7, doxorubicin only treatment; and group 8, without any treatment. Tumor volumes were measured through 18 days. H&E staining, TUNEL assay, and qRT-PCR quantification for delivered miRNAs were performed. <b>Results:</b><i>In vivo</i> results showed that UGMMTD of miRNAs with doxorubicin in groups 1-3 significantly (P<0.05) delayed tumor growth compared to control without any treatment, and doxorubicin only from day 7 to 18. On qRT-PCR, levels of delivered miRNAs were significantly (P<0.05) higher in groups 1-3 upon UGMMTD treatment compared to controls. TUNEL assay showed that upon UGMMTD, significantly higher levels of apoptotic cell populations were observed in groups 1-3 compared to controls. Toxicity was not observed in various organs of different groups. <b>Conclusions:</b> UGMMTD of miRNA-100/miRNA-122/antimiRNA-10b/antimiRNA-21 combination improved therapeutic outcome of doxorubicin chemotherapy in mouse models of HCC by substantial inhibition of tumor growth and significant increase in apoptotic index.</p>","PeriodicalId":36934,"journal":{"name":"Nanotheranostics","volume":"6 1","pages":"62-78"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8671967/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9207100","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":"Advances in Surface Enhanced Raman Spectroscopy for <i>in Vivo</i> Imaging in Oncology.","authors":"Kenry,&nbsp;Fay Nicolson,&nbsp;Louise Clark,&nbsp;Sajanlal R Panikkanvalappil,&nbsp;Bohdan Andreiuk,&nbsp;Chrysafis Andreou","doi":"10.7150/ntno.62970","DOIUrl":"https://doi.org/10.7150/ntno.62970","url":null,"abstract":"<p><p>In the last two decades, the application of surface enhanced Raman scattering (SERS) nanoparticles for preclinical cancer imaging has attracted increasing attention. Raman imaging with SERS nanoparticles offers unparalleled sensitivity, providing a platform for molecular targeting, and granting multiplexed and multimodal imaging capabilities. Recent progress has been facilitated not only by the optimization of the SERS contrast agents themselves, but also by the developments in Raman imaging approaches and instrumentation. In this article, we review the principles of Raman scattering and SERS, present advances in Raman instrumentation specific to cancer imaging, and discuss the biological means of ensuring selective <i>in vivo</i> uptake of SERS contrast agents for targeted, multiplexed, and multimodal imaging applications. We offer our perspective on areas that must be addressed in order to facilitate the clinical translation of SERS contrast agents for <i>in vivo</i> imaging in oncology.</p>","PeriodicalId":36934,"journal":{"name":"Nanotheranostics","volume":" ","pages":"31-49"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8671959/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39640923","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":"Ultrasmall gold nanorod-polydopamine hybrids for enhanced photoacoustic imaging and photothermal therapy in second near-infrared window.","authors":"Wonjun Yim, Raina M Borum, Jiajing Zhou, Yash Mantri, Zhuohong Wu, Jingcheng Zhou, Zhicheng Jin, Matthew Creyer, Jesse V Jokerst","doi":"10.7150/ntno.63634","DOIUrl":"10.7150/ntno.63634","url":null,"abstract":"<p><p>Gold nanorods (GNRs) have attracted great interest for photo-mediated biomedicines due to their tunable and high optical absorption, high photothermal conversion efficiency and facile surface modifiability. GNRs that have efficient absorption in second near-infrared (NIR-II) window hold further promise in bio-applications due to low background signal from tissue and deep tissue penetration. However, bare GNRs readily undergo shape deformation (termed as 'melting effect') during the laser illumination losing their unique localized surface plasmon resonance (LSPR) properties, which subsequently leads to PA signal attenuation and decreased photothermal efficiency. Polydopamine (PDA) is a robust synthetic melanin that has broad absorption and high photothermal conversion. Herein, we coated GNRs with PDA to prepare photothermally robust GNR@PDA hybrids for enhanced photo-mediated theranostic agents. Ultrasmall GNRs (SGNRs) and conventional large GNRs (LGNRs) that possess similar LSPR characteristics as well as GNR@PDA hybrids were compared side-by-side in terms of the size-dependent photoacoustic (PA) imaging, photothermal therapy (PTT), and structural stability. <i>In vitro</i> experiments further demonstrated that SGNR@PDA showed 95% ablation of SKOV3 ovarian cancer cells, which is significantly higher than that of LGNRs (66%) and SGNRs (74%). Collectively, our PDA coating strategy represents a rational design for enhanced PA imaging and efficient PTT via a nanoparticle, i.e., nanotheranostics.</p>","PeriodicalId":36934,"journal":{"name":"Nanotheranostics","volume":"6 1","pages":"79-90"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8671965/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10320614","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":"Flavin Adenine Dinucleotide (FAD) Pegylated (PEG)-Complexes: Proof of Concept (PoC) of theranostic tool on a Murine Breast Cancer Model.","authors":"Celia Arib,&nbsp;Hui Liu,&nbsp;Qiqian Liu,&nbsp;Anne-Marie Cieutat,&nbsp;Didier Paleni,&nbsp;Xiaowu Li,&nbsp;Jolanda Spadavecchia","doi":"10.7150/ntno.63496","DOIUrl":"https://doi.org/10.7150/ntno.63496","url":null,"abstract":"<p><p>Flavin adenine dinucleotide (FAD) plays a key role in an extensive range of cellular oxidation-reduction reactions, which is engaged in metabolic pathways. The purpose of this study was to realize pegylated flavins formulation, named FAD and FAD-PEG diacid complex as theranostic tool in cancer therapy. For this objective, a murine breast cancer model, which was induced by mouse-derived4T1 breast cancer cells was studied to assess the therapeutic efficacy of FAD (named NP1) and FAD-PEG diacid complex (named NP2). The cytokines were monitored to evaluate the serum inflammatory factors to develop the blood cell content of different groups of nude mice. The experimental model shows that an intravenous injection of FAD (NP1) can significantly reduce tumour volume, tumour index and thymus index, and decrease neutrophils (NE), monocytes (MO), eosinophils (EO), and basophils (BA). At the same time, the content of IL-1α, IL-12P70, TNF α, IL-1β and IL-6 was significantly reduced, and the content of IL-10 was significantly increased. These results provide the proof-of-concept for FAD as a smart adjuvant for cancer therapy and encourages their further development in the field of Nanomedicine.</p>","PeriodicalId":36934,"journal":{"name":"Nanotheranostics","volume":" ","pages":"175-183"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8671949/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39656490","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":"Graphene Quantum Dots prepared by Electron Beam Irradiation for Safe Fluorescence Imaging of Tumor.","authors":"Honghong Cao, Wei Qi, Xudong Gao, Qiang Wu, Longlong Tian, Wangsuo Wu","doi":"10.7150/ntno.67070","DOIUrl":"10.7150/ntno.67070","url":null,"abstract":"<p><p>Graphene quantum dots (GQD) have attracted much attention due to their unique properties in biomedical application, such as biosensing, imaging, and drug delivering. However, scale preparing red luminescing GQD is still challenging now. Herein, with the help of electron beam irradiation, a simple, rapid, and efficient up-to-down strategy was developed to synthesize GQD with size of 2.75 nm emitting 610 nm luminescence. GQD were further functionalized with polyethylene glycol (PEG) and exhibited good solubility and biocompatibility. The potential <i>in vivo</i> toxicity of PEGylated GQD could completely be eliminated by the clinic cholesterol-lowering drug simvastatin. PEGylated GQD could selectively accumulate in tumor after intravenous injection as a security, reliable and sensitive tumor fluorescence imaging agent. Therefore, this work presented a new method preparing red luminescing GQD for biomedical application.</p>","PeriodicalId":36934,"journal":{"name":"Nanotheranostics","volume":" ","pages":"205-214"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8671948/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39892502","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":"Design and synthesis of gold nanostars-based SERS nanotags for bioimaging applications.","authors":"Bohdan Andreiuk,&nbsp;Fay Nicolson,&nbsp;Louise M Clark,&nbsp;Sajanlal R Panikkanvalappil,&nbsp;Kenry,&nbsp;Mohammad Rashidian,&nbsp;Stefan Harmsen,&nbsp;Moritz F Kircher","doi":"10.7150/ntno.61244","DOIUrl":"10.7150/ntno.61244","url":null,"abstract":"<p><p>Surface-enhanced Raman spectroscopy (SERS) nanotags hold a unique place among bioimaging contrast agents due to their fingerprint-like spectra, which provide one of the highest degrees of detection specificity. However, in order to achieve a sufficiently high signal intensity, targeting capabilities, and biocompatibility, all components of nanotags must be rationally designed and tailored to a specific application. Design parameters include fine-tuning the properties of the plasmonic core as well as optimizing the choice of Raman reporter molecule, surface coating, and targeting moieties for the intended application. This review introduces readers to the principles of SERS nanotag design and discusses both established and emerging protocols of their synthesis, with a specific focus on the construction of SERS nanotags in the context of bioimaging and theranostics.</p>","PeriodicalId":36934,"journal":{"name":"Nanotheranostics","volume":" ","pages":"10-30"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8671966/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39640922","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":"Review on Optical Imaging Techniques for Multispectral Analysis of Nanomaterials.","authors":"Haeni Lee,&nbsp;Jaeheung Kim,&nbsp;Hyung-Hoi Kim,&nbsp;Chang-Seok Kim,&nbsp;Jeesu Kim","doi":"10.7150/ntno.63222","DOIUrl":"10.7150/ntno.63222","url":null,"abstract":"<p><p>Biomedical imaging is an essential tool for investigating biological responses <i>in vivo</i>. Among the several imaging techniques, optical imaging systems with multispectral analysis of nanoparticles have been widely investigated due to their ability to distinguish the substances in biological tissues <i>in vivo</i>. This review article focus on multispectral optical imaging techniques that can provide molecular functional information. We summarize the basic principle of the spectral unmixing technique that enables the delineation of optical chromophores. Then, we explore the principle, typical system configuration, and biomedical applications of the representative optical imaging techniques, which are fluorescence imaging, two-photon microscopy, and photoacoustic imaging. The results in the recent studies show the great potential of the multispectral analysis techniques for monitoring responses of biological systems <i>in vivo</i>.</p>","PeriodicalId":36934,"journal":{"name":"Nanotheranostics","volume":" ","pages":"50-61"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8671957/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39640924","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}