Bioengineering & Translational Medicine最新文献

{"title":"Mpox disease, diagnosis, and point of care platforms","authors":"Nazente Atceken,&nbsp;Ikra Bayaki,&nbsp;Berk Can,&nbsp;Defne Yigci,&nbsp;Savas Tasoglu","doi":"10.1002/btm2.10733","DOIUrl":"10.1002/btm2.10733","url":null,"abstract":"<p>Human Mpox disease (MPX) is an endemic zoonotic disease that develops when patients are infected with the Mpox virus (MPXV). MPXV shares a high level of genetic similarity to other poxviruses and the clinical presentation of MPX is similar to other poxvirus infections which can result in a delay in diagnosis. In addition, the MPXV virus is phylogenetically divided into two different clades which affects the severity of disease. In recent years, there has been an unusual worldwide spread of MPXV, leading to a global public health problem. The most important step in the fight against MPX is rapid, highly specific, and accurate diagnosis. Following the rapid spread of disease in recent years, efforts to develop diagnostic tests have gained momentum. Here, MPX, MPX epidemiology, and MPX diagnostic tests are discussed. Furthermore, biochemical diagnostic tests, molecular diagnostic tests and their development, and point-of-care (PoC) diagnostic applications are reviewed. Molecular diagnostic technologies such as polymerase chain reaction, recombinase polymerase amplification, and loop-mediated isothermal amplification methods that detect MPX are evaluated. Additionally, next-generation combined molecular techniques and their importance in PoC transition are explored.</p>","PeriodicalId":9263,"journal":{"name":"Bioengineering & Translational Medicine","volume":"10 3","pages":""},"PeriodicalIF":6.1,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btm2.10733","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142925001","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ultrasound-mediated mechanical forces activate selective tumor cell apoptosis","authors":"Ajay Tijore,&nbsp;Felix Margadant,&nbsp;Nehal Dwivedi,&nbsp;Leslie Morgan,&nbsp;Mingxi Yao,&nbsp;Anushya Hariharan,&nbsp;Claire Alexandra Zhen Chew,&nbsp;Simon Powell,&nbsp;Glenn Kunnath Bonney,&nbsp;Michael Sheetz","doi":"10.1002/btm2.10737","DOIUrl":"10.1002/btm2.10737","url":null,"abstract":"<p>Recent studies show that tumor cells undergo apoptosis after mechanical stretching, which promotes normal cell growth. Since ultrasound can produce similar sub-cellular mechanical stresses on the nanoscale, here we test the effect of ultrasound-mediated mechanical forces on tumors and normal cell survival. Surprisingly, tumor cells undergo apoptosis through a calpain-dependent mitochondrial pathway that relies upon calcium entry through the mechanosensitive Piezo1 channels. This is a general property of all tumor cell lines tested irrespective of tissue origin, but normal cells are unaffected. In vivo, ultrasound treatment promotes tumor cell killing in a mouse model with invasive CT26 cancer cell subcutaneous tumors and in the chick chorioallantoic membrane (CAM) model with relatively minor damage to chick embryos. Further, patient-derived pancreatic tumor organoids are killed by ultrasound treatment. Because ultrasound-mediated mechanical forces cause apoptosis of tumor cells from many different tissues in different microenvironments, it may offer a safe, non-invasive approach to augment tumor treatments.</p>","PeriodicalId":9263,"journal":{"name":"Bioengineering & Translational Medicine","volume":"10 2","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btm2.10737","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142908219","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Catalpol promotes the generation of cerebral organoids with oRGs through activation of STAT3 signaling","authors":"Yoo-Jung Lee,&nbsp;Byounggook Cho,&nbsp;Daeyeol Kwon,&nbsp;Yunkyung Kim,&nbsp;Saemin An,&nbsp;Soi Kang,&nbsp;Jongpil Kim","doi":"10.1002/btm2.10746","DOIUrl":"10.1002/btm2.10746","url":null,"abstract":"<p>The generation of human cortical organoids containing outer radial glia (oRG) cells is crucial for modeling neocortical development. Here we show that Catalpol, an iridoid glucoside derived from <i>Rehmannia glutinosa</i>, significantly enhances the generation of cerebral organoids with expanded oRG populations and increased neurogenic potential. Catalpol-treated organoids exhibited thicker ventricular zone/subventricular zone (VZ/SVZ) and outer subventricular zone (oSVZ) regions, with increased numbers of SOX2 + HOPX+ and SOX2 + TNC+ oRG cells and elevated expression of oRG markers HOPX and FAM107A. We found that Catalpol promoted oRG generation through non-vertical divisions of ventricular radial glia (vRG) cells, indicating enhanced oRG generation via asymmetrical divisions. Furthermore, we demonstrated that Catalpol augmented oRG cell numbers through activation of the STAT3 signaling pathway. These findings highlight Catalpol's potential in promoting the generation of cerebral organoids with expanded oRG populations and increased neurogenic potential through STAT3 activation, offering new insights into neocortical development modeling.</p>","PeriodicalId":9263,"journal":{"name":"Bioengineering & Translational Medicine","volume":"10 3","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btm2.10746","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142902021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Wearable photobiomodulation halts thyroid cancer growth by leveraging thyroid photosensitivity","authors":"Changrui Zhao,&nbsp;Kun Fu,&nbsp;Jiameng Tian,&nbsp;Tian Long,&nbsp;Jianzhong Song,&nbsp;Siyu Chen,&nbsp;Chang Liu","doi":"10.1002/btm2.10734","DOIUrl":"10.1002/btm2.10734","url":null,"abstract":"<p>With papillary thyroid carcinoma (PTC) rates rising significantly, concerns about conventional treatments like thyroidectomy and radiotherapy highlight the need for non-invasive options. Our study explores photobiomodulation therapy (PBMT), which uses specific light wavelengths to evoke cellular responses in PTC treatment. Our research utilized a custom-designed optical system to investigate PBMT, finding that blue light at a wavelength of 465 nm can safely and effectively inhibit the proliferation of the TPC-1 PTC cell line by inducing cell cycle arrest. Additionally, we developed a wirelessly powered wearable PBMT device, which is equipped with an advanced light delivery system that ensures precise and consistent dosage. This device designed for optimal patient comfort, effectively suppressed tumor growth in mouse models without adverse effects. PBMT indicates thyroid tissue's light responsiveness as a non-visual organ. Our study's innovative approach integrates the disciplines of oncology, biophysics, and medical device technology, thereby advancing the treatment paradigms for PTC. This interdisciplinary bridge not only highlights our groundbreaking findings but also paves the way for future research in cancer therapy and photomedicine.</p>","PeriodicalId":9263,"journal":{"name":"Bioengineering & Translational Medicine","volume":"10 2","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btm2.10734","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142867274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent developments in microwire‐structured intracortical electrode arrays for brain–machine interfaces","authors":"Sorel E. De León, Simon Higham, Young Jun Jung, Wei Tong, David J. Garrett","doi":"10.1002/btm2.10742","DOIUrl":"https://doi.org/10.1002/btm2.10742","url":null,"abstract":"Brain–machine interfaces (BMIs) have experienced remarkable advancements in recent years, marked by multiple companies initiating human trials. Consequently, the interface between the brain and electrodes has become more critical than ever, requiring implanted electrodes to be not only biocompatible and minimally invasive but also capable of remaining functioning in the brain for a lifetime. While significant progress has been made in the manufacturing of intracortical electrodes, challenges persist in ensuring longevity and minimizing tissue damage. Additionally, the reliance on manual labor in fabrication techniques poses obstacles to large‐scale production for commercialization. In this review, we explore recent breakthroughs and obstacles in the fabrication of microwire‐structured electrode arrays, wherein single wires are arranged in an <jats:italic>xy</jats:italic> matrix for cortical penetration. We discuss the impact of various fabrication strategies and materials on implant longevity, as well as the remaining challenges in this field.","PeriodicalId":9263,"journal":{"name":"Bioengineering & Translational Medicine","volume":"24 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142867276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The potential of low-intensity pulsed ultrasound to apply the long-term ovary protection from injury induced by 4-vinylcyclohexene diepoxide through inhibiting granulosa cell apoptosis","authors":"Juan Deng,&nbsp;Juan Qin,&nbsp;Guolin Song,&nbsp;Chenghai Li,&nbsp;Wentao Tang,&nbsp;Yilin Tang,&nbsp;Xinfang Xiao,&nbsp;Liu Wu,&nbsp;Sicheng He,&nbsp;Yiqing Zhou,&nbsp;Junfen Li,&nbsp;Yan Wang","doi":"10.1002/btm2.10744","DOIUrl":"10.1002/btm2.10744","url":null,"abstract":"<p>The potential of low-intensity pulsed ultrasound (LIPUS) in regulating ovarian function has been demonstrated; however, there is a lack of scientific evidence regarding the long-term efficacy of LIPUS in treating ovarian injury and understanding its regulatory mechanisms. In this study, 4-vinylcyclohexene diepoxide (VCD) was used to induce ovarian injury in rats, and LIPUS was applied to target the damaged ovarian tissues. The research aimed to investigate the long-term protective effect of LIPUS against ovum toxicity induced by VCD and elucidate the associated molecular mechanisms. During the experiment, HE staining was employed for observing the morphology and structure of the ovary, while protein sequencing was utilized for identifying and confirming the molecular mechanism through which LIPUS restores the damaged ovarian structure. The long-term effectiveness of LIPUS in protecting against ovarian injury was evaluated through ELISA, estrous cycle monitoring, fertility testing, and behavioral analysis. The results indicated that LIPUS effectively restored the structure of damaged ovaries. Both in vivo and in vitro studies revealed that this protective effect may be attributed to LIPUS inhibiting apoptosis of ovarian granulosa cells (GCs) by regulating Daxx-mediated ASK1/JNK signaling pathway. Subsequent functional tests demonstrated significant improvements in sex hormone secretion and regulation of estrous cycle within 6 cycles following LIPUS treatment. Additionally, there was a notable increase in offspring numbers after mating. Behavioral analysis revealed that LIPUS effectively alleviated menopausal symptoms resulting from ovarian injury including mood fluctuations, cognitive behavior changes, and reduced muscle excitability levels. These findings suggest that beneficial effects of LIPUS may help reduce VCD-induced ovarian damage with long-term efficacy.</p>","PeriodicalId":9263,"journal":{"name":"Bioengineering & Translational Medicine","volume":"10 3","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btm2.10744","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142841428","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nanoparticle contrast-enhanced computed tomography and magnetic resonance imaging of vascularization of a subcutaneous niche for islet transplantation","authors":"Simone Capuani,&nbsp;Jocelyn Nikita Campa-Carranza,&nbsp;Nathanael Hernandez,&nbsp;Renuka T. R. Menon,&nbsp;Rohan Bhavane,&nbsp;Gabrielle E. Rome,&nbsp;Laxman Devkota,&nbsp;Ketan B. Ghaghada,&nbsp;Ananth V. Annapragada,&nbsp;Corrine Ying Xuan Chua,&nbsp;Andrew A. Badachhape,&nbsp;Alessandro Grattoni","doi":"10.1002/btm2.10740","DOIUrl":"10.1002/btm2.10740","url":null,"abstract":"<p>Revascularization plays a critical role in the successful engraftment of transplanted pancreatic islets, which are inherently rich in capillaries to meet their high metabolic demands. Innovative islet encapsulation strategies such as the NICHE (neovascularized implantable cell homing and encapsulation), generate a prevascularized transplantation site that allows for direct integration of the graft with the systemic circulation. Timing the transplantation is key to maximizing islet engraftment and survival, especially in diabetic individuals, who exhibit impaired wound healing. Therefore, in this study, we explored different methods to assess vascular development within NICHE in vivo in a non-invasive fashion. We effectively tracked neoangiogenesis using nanoparticle contrast-enhanced computed tomography (nCECT), observing a steady increase in vascularization over an 8-week period, which was confirmed histologically. Next, we estimated relative vascularization changes via T2 mapping with magnetic resonance imaging (MRI) before and after islet transplantation. On the first day post-transplantation, we measured a slight decrease in T2 values followed by a significant increase by day 14 attributable to islet revascularization. Our findings underscore the potential of non-invasive imaging techniques to provide insightful information on the readiness of the transplant site within cell encapsulation systems to support cell graft transplantation.</p>","PeriodicalId":9263,"journal":{"name":"Bioengineering & Translational Medicine","volume":"10 3","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btm2.10740","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142823160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessing the impact of airborne particulate pollution on human skin utilizing a novel human skin equivalent containing MUTZ-3-derived Langerhans cells","authors":"Amy Simpson,&nbsp;Teresa DiColandrea,&nbsp;Stefan Przyborski","doi":"10.1002/btm2.10738","DOIUrl":"10.1002/btm2.10738","url":null,"abstract":"<p>Air pollution is an exogenous stressor known to have a detrimental impact on skin health through the induction of inflammation; however, the direct effect of topical pollution exposure is still being elucidated. Human skin equivalents (HSE) aim to reproduce in vitro the structure and function of the native skin tissue. However, HSEs typically lack skin-resident immune cells, which could play a key role in the inflammatory response induced by pollution exposure. We outline the development of a HSE-containing MUTZ-3-derived Langerhans cells (MUTZ-3-LCs), which show dendritic morphology and Langerhans cell marker expression. We demonstrated that HSE-containing MUTZ-3-LC have lower basal levels of proinflammatory cytokines, but topical stimulation with allergens and irritant compounds induced a greater inflammatory response in these models compared to HSE without immune cells. To study the effect of pollution, we created a technique to apply diesel particulate matter (DPM) to HSEs. Though our microscopic analysis demonstrated that DPM does not penetrate the stratum corneum, we showed that DPM did induce production of proinflammatory cytokines, but notably only in HSEs containing MUTZ-3-LCs. These data suggest that topical exposure to air pollution can induce cutaneous inflammation and that skin-resident immune cells contribute to this response. This highlights the significance of immune-competent HSEs to the study of exogenous stressors in vitro.</p>","PeriodicalId":9263,"journal":{"name":"Bioengineering & Translational Medicine","volume":"10 2","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btm2.10738","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142823164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reduced glutathione enhances adipose tissue-derived mesenchymal stem cell engraftment efficiency for liver fibrosis by targeting TGFβ1/SMAD3/NOX4 pathway","authors":"Shaoxiong Yu,&nbsp;Yingchao Wang,&nbsp;Yingjun Shi,&nbsp;Saihua Yu,&nbsp;Bixing Zhao,&nbsp;Naishun Liao,&nbsp;Xiaolong Liu","doi":"10.1002/btm2.10735","DOIUrl":"10.1002/btm2.10735","url":null,"abstract":"<p>Reduced glutathione (GSH) could reduce oxidative stress to improve adipose tissue-derived mesenchymal stem cell (ADSC) engraftment efficiency in vivo. However, the underlying mechanisms remain unclear. Our goal is to investigate whether GSH enhances ADSC engraftment through targeting the TGFβ/SMAD3/NOX4 pathway. Liver fibrotic male mice were administrated GSH, setanaxib (STX), and SIS3 during ADSC transplantation. ADSC engraftment efficiency and reactive oxygen species (ROS) level were detected both in vivo and ex vivo. Biochemical analysis was used to analyze the content of superoxide and nicotinamide adenine dinucleotide phosphate oxidases (NOXs) in liver tissues. Immunohistochemistry and western blotting were used to examine the protein level of NOX1, NOX2, NOX4, transforming growth factor-β1 (TGFβ1), SMAD3, and p-SMAD3 in liver tissues. Additionally, the therapeutic efficacy of the ADSC transplantation was further investigated. We found that GSH significantly improved ADSC engraftment efficiency, which was closely related to the reduced ROS generation in liver tissues. However, the enhanced cell engraftment was abolished after the combined treatment with STX or SIS3. GSH could effectively reduce superoxide and NOXs content, and selectively inhibit NOX4 expression in liver tissues. The co-localization results showed that GSH could reduce NOX4 expressed in activated hepatic stellate cells. Mechanistically, GSH down-regulated TGFβ/SMAD3 signaling. More importantly, GSH enhanced the therapeutic efficacy of ADSC therapy in liver fibrotic mice. Taken together, GSH could improve the engraftment efficiency of ADSCs in liver fibrosis by targeting TGFβ1/SMAD3/NOX4 signaling pathway, which provides a new theoretical basis for GSH enhancing ADSC engraftment efficiency in liver diseases.</p>","PeriodicalId":9263,"journal":{"name":"Bioengineering & Translational Medicine","volume":"10 2","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btm2.10735","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142804738","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Green/red fluorescent protein disrupting drugs for real-time permeability tracking in three-dimensional tumor spheroids","authors":"Maytal Avrashami,&nbsp;Danna Niezni,&nbsp;Dana Meron Azagury,&nbsp;Hagit Sason,&nbsp;Yosi Shamay","doi":"10.1002/btm2.10731","DOIUrl":"10.1002/btm2.10731","url":null,"abstract":"<p>Three-dimensional (3D) spheroid models offer a more physiologically relevant and complex environment compared to traditional two-dimensional cultures, making them a promising tool for studying tumor biology and drug response. However, these models often face challenges in real-time monitoring of drug diffusion, penetration, and target engagement, limiting their predictive power for in vivo and clinical outcomes. This study introduces a novel approach for real-time tracking of drug permeability using small molecule drugs with GFP/RFP-disrupting properties that correlate with their efficacy. We developed a reproducible 3D spheroid model with various cancer cell lines expressing GFP/RFP for efficient drug screening. Through screening over 20 FDA-approved enzyme inhibitors, we identified three covalent kinase inhibitors—osimertinib, afatinib, and neratinib—that irreversibly disrupt GFP and RFP fluorescence. Our results reveal distinct drug diffusion and penetration profiles within GFP/RFP-expressing spheroids, varying with drug concentration and formulation, and correlating with clinical volume of distribution (Vd) values. Additionally, we demonstrate that our approach is useful for evaluating different drug formulations as well as screening penetration enhancers for solid tumors. These findings offer a valuable 3D model for studying kinetics of drug permeability and efficacy in tumor-like environments, with potential implications for drug delivery research and formulation development.</p>","PeriodicalId":9263,"journal":{"name":"Bioengineering & Translational Medicine","volume":"10 3","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/btm2.10731","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142796884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}