Neuropathology and Applied Neurobiology最新文献

{"title":"Motor neuron involvement threatens survival in spinocerebellar ataxia type 1.","authors":"Giulia Coarelli, Maya Tchikviladzé, Pauline Dodet, Isabelle Arnulf, Perrine Charles, Frederic Tankeré, Thomas Similowski, Danielle Seilhean, Alexis Brice, Charles Duyckaerts, Alexandra Durr","doi":"10.1111/nan.12897","DOIUrl":"https://doi.org/10.1111/nan.12897","url":null,"abstract":"Sorbonne Université, Paris Brain Institute (ICM Institut du Cerveau), INSERM, CNRS, Assistance Publique-Hôpitaux de Paris (AP-HP), Paris, France Department of Genetics, Pitié-Salpêtrière Charles-Foix University Hospital, Assistance Publique – Hôpitaux de Paris (AP-HP), Paris, 75013, France Sleep Disorders Unit, Pitié-Salpêtrière University Hospital, AP-HP, Paris, France; ICM, Sorbonne Université, Inserm U 1127, CNRS UMR, Paris, 7225, France Department of Otolaryngology-Head and Neck Surgery, Pitié-Salpêtrière Charles-Foix University Hospital, Assistance Publique – Hôpitaux de Paris (AP-HP), Sorbonne Université, Paris, 75013, France Département R3S (Respiration, Réanimation, Réhabilitation respiratoire, Sommeil), Sorbonne Université, INSERM, UMRS1158 Neurophysiologie Respiratoire Expérimentale et Clinique; AP-HP, Groupe Hospitalier Universitaire APHP-Sorbonne Université, site Pitié-Salpêtrière, Paris, F-75013, France Laboratoire de Neuropathologie R. Escourolle, Pitié-Salpêtrière Charles-Foix University Hospital, Assistance Publique – Hôpitaux de Paris (AP-HP), Sorbonne Université, Paris, 75013, France","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"49 2","pages":"e12897"},"PeriodicalIF":5.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9366537","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":"Muscleblind-like 2 knockout shifts adducin 1 isoform expression and alters dendritic spine dynamics of cortical neurons during brain development.","authors":"Chia-Wei Huang, Kuang-Yung Lee, Peng-Tzu Lin, Fang-Shin Nian, Haw-Yuan Cheng, Chien-Hui Chang, Cheng-Yen Liao, Yen-Lin Su, Carol Seah, Ching Li, Yu-Fu Chen, Mei-Hsuan Lee, Jin-Wu Tsai","doi":"10.1111/nan.12890","DOIUrl":"https://doi.org/10.1111/nan.12890","url":null,"abstract":"Muscleblind‐like 2 (MBNL2) plays a crucial role in regulating alternative splicing during development and mouse loss of MBNL2 recapitulates brain phenotypes in myotonic dystrophy (DM). However, the mechanisms underlying DM neuropathogenesis during brain development remain unclear. In this study, we aim to investigate the impact of MBNL2 elimination on neuronal development by Mbnl2 conditional knockout (CKO) mouse models.","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"49 2","pages":"e12890"},"PeriodicalIF":5.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9423814","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":"Soluble amyloid-β dimers are resistant to amyloid-β prion conversion in vivo suggesting antiprion properties.","authors":"Else F van Gerresheim, Andreas Müller-Schiffmann, Sandra Schäble, Bastijn Koopmans, Maarten Loos, Carsten Korth","doi":"10.1111/nan.12895","DOIUrl":"https://doi.org/10.1111/nan.12895","url":null,"abstract":"According to the instructions for authors, short communications should be published without abstract.","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"49 2","pages":"e12895"},"PeriodicalIF":5.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9721544","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":"TDP-43 pathology and functional deficits in wild-type and ALS/FTD mutant cyclin F mouse models.","authors":"Annika van Hummel, Miheer Sabale, Magdalena Przybyla, Julia van der Hoven, Gabriella Chan, Astrid F Feiten, Roger S Chung, Lars M Ittner, Yazi D Ke","doi":"10.1111/nan.12902","DOIUrl":"10.1111/nan.12902","url":null,"abstract":"<p><strong>Aims: </strong>Amyotrophic lateral sclerosis (ALS) is characterised by a progressive loss of upper and lower motor neurons leading to muscle weakness and eventually death. Frontotemporal dementia (FTD) presents clinically with significant behavioural decline. Approximately 10% of cases have a known family history, and disease-linked mutations in multiple genes have been identified in FTD and ALS. More recently, ALS and FTD-linked variants have been identified in the CCNF gene, which accounts for an estimated 0.6% to over 3% of familial ALS cases.</p><p><strong>Methods: </strong>In this study, we developed the first mouse models expressing either wild-type (WT) human CCNF or its mutant pathogenic variant S621G to recapitulate key clinical and neuropathological features of ALS and FTD linked to CCNF disease variants. We expressed human CCNF WT or CCNF^S621G throughout the murine brain by intracranial delivery of adeno-associated virus (AAV) to achieve widespread delivery via somatic brain transgenesis.</p><p><strong>Results: </strong>These mice developed behavioural abnormalities, similar to the clinical symptoms of FTD patients, as early as 3 months of age, including hyperactivity and disinhibition, which progressively deteriorated to include memory deficits by 8 months of age. Brains of mutant CCNF_S621G mice displayed an accumulation of ubiquitinated proteins with elevated levels of phosphorylated TDP-43 present in both CCNF_WT and mutant CCNF_S621G mice. We also investigated the effects of CCNF expression on interaction targets of CCNF and found elevated levels of insoluble splicing factor proline and glutamine-rich (SFPQ). Furthermore, cytoplasmic TDP-43 inclusions were found in both CCNF_WT and mutant CCNF_S621G mice, recapitulating the key hallmark of FTD/ALS pathology.</p><p><strong>Conclusions: </strong>In summary, CCNF expression in mice reproduces clinical presentations of ALS, including functional deficits and TDP-43 neuropathology with altered CCNF-mediated pathways contributing to the pathology observed.</p>","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"49 2","pages":"e12902"},"PeriodicalIF":5.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10946706/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9360740","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":"DNA methylation-based classification of glioneuronal tumours synergises with histology and radiology to refine accurate molecular stratification.","authors":"Thomas J Stone, Kshitij Mankad, Ai Peng Tan, Wajanat Jan, Jessica C Pickles, Maria Gogou, Jane Chalker, Iwona Slodkowska, Emily Pang, Mark Kristiansen, Gaganjit K Madhan, Leysa Forrest, Deborah Hughes, Eleni Koutroumanidou, Talisa Mistry, Olumide Ogunbiyi, Saira W Ahmed, J Helen Cross, Mike Hubank, Darren Hargrave, Thomas S Jacques","doi":"10.1111/nan.12894","DOIUrl":"10.1111/nan.12894","url":null,"abstract":"<p><strong>Aims: </strong>Glioneuronal tumours (GNTs) are poorly distinguished by their histology and lack robust diagnostic indicators. Previously, we showed that common GNTs comprise two molecularly distinct groups, correlating poorly with histology. To refine diagnosis, we constructed a methylation-based model for GNT classification, subsequently evaluating standards for molecular stratification by methylation, histology and radiology.</p><p><strong>Methods: </strong>We comprehensively analysed methylation, radiology and histology for 83 GNT samples: a training cohort of 49, previously classified into molecularly defined groups by genomic profiles, plus a validation cohort of 34. We identified histological and radiological correlates to molecular classification and constructed a methylation-based support vector machine (SVM) model for prediction. Subsequently, we contrasted methylation, radiological and histological classifications in validation GNTs.</p><p><strong>Results: </strong>By methylation clustering, all training and 23/34 validation GNTs segregated into two groups, the remaining 11 clustering alongside control cortex. Histological review identified prominent astrocytic/oligodendrocyte-like components, dysplastic neurons and a specific glioneuronal element as discriminators between groups. However, these were present in only a subset of tumours. Radiological review identified location, margin definition, enhancement and T2 FLAIR-rim sign as discriminators. When validation GNTs were classified by SVM, 22/23 classified correctly, comparing favourably against histology and radiology that resolved 17/22 and 15/21, respectively, where data were available for comparison.</p><p><strong>Conclusions: </strong>Diagnostic criteria inadequately reflect glioneuronal tumour biology, leaving a proportion unresolvable. In the largest cohort of molecularly defined glioneuronal tumours, we develop molecular, histological and radiological approaches for biologically meaningful classification and demonstrate almost all cases are resolvable, emphasising the importance of an integrated diagnostic approach.</p>","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"49 2","pages":"e12894"},"PeriodicalIF":5.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10946721/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9367728","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":"Schwann cells and myelin in human peripheral nerve: Major protein components vary with age, axon size and pathology.","authors":"Alan Pestronk,&nbsp;Robert E Schmidt,&nbsp;Robert Bucelli,&nbsp;Julia Sim","doi":"10.1111/nan.12898","DOIUrl":"https://doi.org/10.1111/nan.12898","url":null,"abstract":"<p><strong>Aims: </strong>We examined major protein components of Schwann cells (SCs) and myelin in normal and diseased human peripheral nerves.</p><p><strong>Methods: </strong>We evaluated distributions of neural cell adhesion molecule (NCAM), P0 protein (P0) and myelin basic protein (MBP) in frozen sections of 98 sural nerves.</p><p><strong>Results: </strong>Non-myelinating SC in normal adults contained NCAM, but not P0 or MBP. With chronic axon loss, SC without associated axons (Büngner band cells) often co-stained for both NCAM and P0. Onion bulb cells also co-stained for both P0 and NCAM. Infants had many SC with MBP but no P0. All myelin sheaths contained P0. Myelin around large, and some intermediate-sized, axons co-stained for both MBP and P0. Myelin on other intermediate-sized axons had P0, but no MBP. Regenerated axons often had sheaths with MBP, P0 and some NCAM. During active axon degeneration, myelin ovoids often co-stained for MBP, P0 and NCAM. Demyelinating neuropathy patterns included SC (NCAM) loss, and myelin with abnormally distributed, or reduced, P0.</p><p><strong>Conclusions: </strong>Peripheral nerve SC and myelin have varied molecular phenotypes, related to age, axon size and nerve pathology. In normal adult peripheral nerve, myelin has two different patterns of molecular composition. MBP is mostly absent from myelin around a population of intermediate-sized axons, whereas P0 is present in myelin around all axons. Denervated SCs have a molecular signature that differs from normal SC types. With acute denervation, SCs may stain for both NCAM and MBP. Chronically denervated SCs often stain for both NCAM and P0.</p>","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"49 2","pages":"e12898"},"PeriodicalIF":5.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9690854","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":"Fibrocartilaginous cerebral and spinal emboli: A report of two cases with histopathological confirmation.","authors":"Fouzia Ziad,&nbsp;David Wang,&nbsp;Andrew Chancellor,&nbsp;Zakier Hussain,&nbsp;Adam El-Dieb,&nbsp;Sanjeevan Pasupati,&nbsp;Thomas Robertson","doi":"10.1111/nan.12896","DOIUrl":"https://doi.org/10.1111/nan.12896","url":null,"abstract":"Department of Pathology, Waikato Hospital, Hamilton, New Zealand Department of Medicine, Auckland City Hospital, Auckland, New Zealand Department of Neurology, Tauranga Hospital, Tauranga, New Zealand Department of Neurosurgery, Waikato Hospital, Hamilton, New Zealand Department of Radiology, Tauranga Hospital, Tauranga, New Zealand Department of Cardiology, Waikato Hospital, Hamilton, New Zealand Pathology Queensland, Royal Brisbane and Women’s Hospital, Brisbane, Queensland, Australia","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"49 2","pages":"e12896"},"PeriodicalIF":5.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9721546","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":"Patient-derived xenograft mouse models to investigate tropism to the central nervous system and retina of primary and secondary central nervous system lymphoma.","authors":"Lisa Kristina Isbell,&nbsp;Cordula Tschuch,&nbsp;Soroush Doostkam,&nbsp;Silvia Waldeck,&nbsp;Geoffroy Andrieux,&nbsp;Khalid Shoumariyeh,&nbsp;Dorothee Lenhard,&nbsp;Hans Eckart Schaefer,&nbsp;Peter Christoph Reinacher,&nbsp;Ingrid Bartsch,&nbsp;Milena Pantic,&nbsp;Janaki Manoja Vinnakota,&nbsp;Vinodh Kakkassery,&nbsp;Elisabeth Schorb,&nbsp;Florian Scherer,&nbsp;Anna Verena Frey,&nbsp;Melanie Boerries,&nbsp;Gerald Illerhaus,&nbsp;Justus Duyster,&nbsp;Julia Schueler,&nbsp;Nikolas von Bubnoff","doi":"10.1111/nan.12899","DOIUrl":"https://doi.org/10.1111/nan.12899","url":null,"abstract":"<p><strong>Aims: </strong>How and why lymphoma cells home to the central nervous system and vitreoretinal compartment in primary diffuse large B-cell lymphoma of the central nervous system remain unknown. Our aim was to create an in vivo model to study lymphoma cell tropism to the central nervous system.</p><p><strong>Methods: </strong>We established a patient-derived central nervous system lymphoma xenograft mouse model and characterised xenografts derived from four primary and four secondary central nervous system lymphoma patients using immunohistochemistry, flow cytometry and nucleic acid sequencing technology. In reimplantation experiments, we analysed dissemination patterns of orthotopic and heterotopic xenografts and performed RNA sequencing of different involved organs to detect differences at the transcriptome level.</p><p><strong>Results: </strong>We found that xenografted primary central nervous system lymphoma cells home to the central nervous system and eye after intrasplenic transplantation, mimicking central nervous system and primary vitreoretinal lymphoma pathology, respectively. Transcriptomic analysis revealed distinct signatures for lymphoma cells in the brain in comparison to the spleen as well as a small overlap of commonly regulated genes in both primary and secondary central nervous system lymphoma.</p><p><strong>Conclusion: </strong>This in vivo tumour model preserves key features of primary and secondary central nervous system lymphoma and can be used to explore critical pathways for the central nervous system and retinal tropism with the goal to find new targets for novel therapeutic approaches.</p>","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"49 2","pages":"e12899"},"PeriodicalIF":5.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9367743","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":"A standardised protocol for blood and cerebrospinal fluid collection and processing for biomarker research in ataxia.","authors":"Magda M Santana, Laetitia S Gaspar, Maria M Pinto, Patrick Silva, Diana Adão, Dina Pereira, Joana Afonso Ribeiro, Inês Cunha, Jeannette Huebener-Schmid, Mafalda Raposo, Ana F Ferreira, Jennifer Faber, Sandra Kuhs, Hector Garcia-Moreno, Kathrin Reetz, Andreas Thieme, Jon Infante, Bart P C van de Warrenburg, Paola Giunti, Olaf Riess, Ludger Schöls, Manuela Lima, Thomas Klockgether, Cristina Januário, Luís Pereira de Almeida","doi":"10.1111/nan.12892","DOIUrl":"10.1111/nan.12892","url":null,"abstract":"<p><p>The European Spinocerebellar Ataxia Type 3/Machado-Joseph Disease Initiative (ESMI) is a consortium established with the ambition to set up the largest European longitudinal trial-ready cohort of Spinocerebellar Ataxia Type 3/Machado-Joseph Disease (SCA3/MJD), the most common autosomal dominantly inherited ataxia worldwide. A major focus of ESMI has been the identification of SCA3/MJD biomarkers to enable future interventional studies. As biosample collection and processing variables significantly impact the outcomes of biomarkers studies, biosampling procedures standardisation was done previously to study visit initiation. Here, we describe the ESMI consensus biosampling protocol, developed within the scope of ESMI, that ultimately might be translated to other neurodegenerative disorders, particularly ataxias, being the first step to protocol harmonisation in the field.</p>","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"49 2","pages":"e12892"},"PeriodicalIF":5.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10947376/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9423824","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":"Paths to hippocampal damage in neuromyelitis optica spectrum disorders.","authors":"Mona Zakani, Magdalini Nigritinou, Markus Ponleitner, Yoshiki Takai, Daniel Hofmann, Sophie Hillebrand, Romana Höftberger, Jan Bauer, Balint Lasztoczi, Tatsuro Misu, Gregor Kasprian, Paulus Rommer, Monika Bradl","doi":"10.1111/nan.12893","DOIUrl":"10.1111/nan.12893","url":null,"abstract":"<p><strong>Aims: </strong>Many patients with neuromyelitis optica spectrum disorders (NMOSD) suffer from cognitive impairment affecting memory, processing speed and attention and suffer from depressive symptoms. Because some of these manifestations could trace back to the hippocampus, several magnetic resonance imaging (MRI) studies have been performed in the past, with a number of groups describing volume loss of the hippocampus in NMOSD patients, whereas others did not observe such changes. Here, we addressed these discrepancies.</p><p><strong>Methods: </strong>We performed pathological and MRI studies on the hippocampi of NMOSD patients, combined with detailed immunohistochemical analysis of hippocampi from experimental models of NMOSD.</p><p><strong>Results: </strong>We identified different pathological scenarios for hippocampal damage in NMOSD and its experimental models. In the first case, the hippocampus was compromised by the initiation of astrocyte injury in this brain region and subsequent local effects of microglial activation and neuronal damage. In the second case, loss of hippocampal volume was seen by MRI in patients with large tissue-destructive lesions in the optic nerves or the spinal cord, and the pathological work-up of tissue derived from a patient with such lesions revealed subsequent retrograde neuronal degeneration affecting different axonal tracts and neuronal networks. It remains to be seen whether remote lesions and associated retrograde neuronal degeneration on their own are sufficient to cause extensive volume loss of the hippocampus, or whether they act in concert with small astrocyte-destructive, microglia-activating lesions in the hippocampus that escape detection by MRI, either due to their small size or due to the chosen time window for examination.</p><p><strong>Conclusions: </strong>Different pathological scenarios can culminate in hippocampal volume loss in NMOSD patients.</p>","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"49 2","pages":"e12893"},"PeriodicalIF":4.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10947283/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9721106","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}