Correction: Verification in the Early Stages of the COVID-19 Pandemic: Sentiment Analysis of Japanese Twitter Users.

[This corrects the article DOI: 10.2196/37881.].

C5a-C5AR1 axis as a potential trigger of the rupture of intracranial aneurysms.

Recent studies have indicated the involvement of neutrophil-mediated inflammatory responses in the process leading to intracranial aneurysm (IA) rupture. Receptors mediating neutrophil recruitment could thus be therapeutic targets of unruptured IAs. In this study, complement C5a receptor 1 (C5AR1) was picked up as a candidate that may cause neutrophil-dependent inflammation in IA lesions from comprehensive gene expression profile data acquired from rat and human samples. The induction of C5AR1 in IA lesions was confirmed by immunohistochemistry; the up-regulations of C5AR1/C5ar1 stemmed from infiltrated neutrophils, which physiologically express C5AR1/C5ar1, and adventitial fibroblasts that induce C5AR1/C5ar1 in human/rat IA lesions. In in vitro experiments using NIH/3T3, a mouse fibroblast-like cell line, induction of C5ar1 was demonstrated by starvation or pharmacological inhibition of mTOR signaling by Torin1. Immunohistochemistry and an experiment in a cell-free system using recombinant C5 protein and recombinant Plasmin indicated that the ligand of C5AR1, C5a, could be produced through the enzymatic digestion by Plasmin in IA lesions. In conclusion, we have identified a potential contribution of the C5a-C5AR1 axis to neutrophil infiltration as well as inflammatory responses in inflammatory cells and fibroblasts of IA lesions. This cascade may become a therapeutic target to prevent the rupture of IAs.

Time-of-flight and black-blood MRI to study intracranial arteries in rats.

Intracranial aneurysms (IAs) are usually incidentally discovered by magnetic resonance imaging (MRI). Once discovered, the risk associated with their treatment must be balanced with the risk of an unexpected rupture. Although clinical observations suggest that the detection of contrast agent in the aneurysm wall using a double-inversion recovery black-blood (BB) sequence may point to IA wall instability, the exact meaning of this observation is not understood. Validation of reliable diagnostic markers of IA (in)stability is of utmost importance to deciding whether to treat or not an IA. To longitudinally investigate IA progression and enhance our understanding of this devastating disease, animal models are of great help. The aim of our study was to improve a three-dimensional (3D)-time-of-flight (TOF) sequence and to develop a BB sequence on a standard preclinical 3-T MRI unit to investigate intracranial arterial diseases in rats. We showed that our 3D-TOF sequence allows reliable measurements of intracranial artery diameters, inter-artery distances, and angles between arteries and that our BB sequence enables us to visualize intracranial arteries. We report the first BB-MRI sequence to visualize intracranial arteries in rats using a preclinical 3-T MRI unit. This sequence could be useful for a large community of researchers working on intracranial arterial diseases.Relevance statement We developed a black-blood MRI sequence to study vessel wall enhancement in rats with possible application to understanding IAs instability and finding reliable markers for clinical decision-making.Key points• Reliable markers of aneurysm stability are needed for clinical decision.• Detection of contrast enhancement in the aneurysm wall may be associated with instability.• We developed a black-blood MRI sequence in rats to be used to study vessel wall enhancement of IAs.

Application of deep neural survival networks to the development of risk prediction models for diabetes mellitus, hypertension, and dyslipidemia.

OBJECTIVES: : Although numerous risk prediction models have been proposed, few such models have been developed using neural network-based survival analysis. We developed risk prediction models for three cardiovascular disease risk factors (diabetes mellitus, hypertension, and dyslipidemia) among a working-age population in Japan using DeepSurv, a deep feed-forward neural network. METHODS: : Data were obtained from the Japan Epidemiology Collaboration on Occupational Health Study. A total of 51 258, 44 197, and 31 452 individuals were included in the development of risk models for diabetes mellitus, hypertension, and dyslipidemia, respectively; two-thirds of whom were used to develop prediction models, and the rest were used to validate the models. We compared the performances of DeepSurv-based models with those of prediction models based on the Cox proportional hazards model. RESULTS: : The area under the receiver-operating characteristic curve was 0.878 [95% confidence interval (CI) = 0.864-0.892] for diabetes mellitus, 0.835 (95% CI = 0.826-0.845) for hypertension, and 0.826 (95% CI = 0.817-0.835) for dyslipidemia. Compared with the Cox proportional hazards-based models, the DeepSurv-based models had better reclassification performance [diabetes mellitus: net reclassification improvement (NRI) = 0.474, P  ≤ 0.001; hypertension: NRI = 0.194, P  ≤ 0.001; dyslipidemia: NRI = 0.397, P  ≤ 0.001] and discrimination performance [diabetes mellitus: integrated discrimination improvement (IDI) = 0.013, P  ≤ 0.001; hypertension: IDI = 0.007, P  ≤ 0.001; and dyslipidemia: IDI = 0.043, P  ≤ 0.001]. CONCLUSION: : This study suggests that DeepSurv has the potential to improve the performance of risk prediction models for cardiovascular disease risk factors.

Spatially Resolved Tumor Microenvironment Predicts Treatment Outcomes in Relapsed/Refractory Hodgkin Lymphoma.

PURPOSE: About a third of patients with relapsed or refractory classic Hodgkin lymphoma (r/r CHL) succumb to their disease after high-dose chemotherapy followed by autologous stem-cell transplantation (HDC/ASCT). Here, we aimed to describe spatially resolved tumor microenvironment (TME) ecosystems to establish novel biomarkers associated with treatment failure in r/r CHL. PATIENTS AND METHODS: We performed imaging mass cytometry (IMC) on 71 paired primary diagnostic and relapse biopsies using a marker panel specific to CHL biology. For each cell type in the TME, we calculated a spatial score measuring the distance of nearest neighbor cells to the malignant Hodgkin Reed Sternberg cells within the close interaction range. Spatial scores were used as features in prognostic model development for post-ASCT outcomes. RESULTS: Highly multiplexed IMC data revealed shared TME patterns in paired diagnostic and early r/r CHL samples, whereas TME patterns were more divergent in pairs of diagnostic and late relapse samples. Integrated analysis of IMC and single-cell RNA sequencing data identified unique architecture defined by CXCR5+ Hodgkin and Reed Sternberg (HRS) cells and their strong spatial relationship with CXCL13+ macrophages in the TME. We developed a prognostic assay (RHL4S) using four spatially resolved parameters, CXCR5+ HRS cells, PD1+CD4+ T cells, CD68+ tumor-associated macrophages, and CXCR5+ B cells, which effectively separated patients into high-risk versus low-risk groups with significantly different post-ASCT outcomes. The RHL4S assay was validated in an independent r/r CHL cohort using a multicolor immunofluorescence assay. CONCLUSION: We identified the interaction of CXCR5+ HRS cells with ligand-expressing CXCL13+ macrophages as a prominent crosstalk axis in relapsed CHL. Harnessing this TME biology, we developed a novel prognostic model applicable to r/r CHL biopsies, RHL4S, opening new avenues for spatial biomarker development.

Verification in the Early Stages of the COVID-19 Pandemic: Sentiment Analysis of Japanese Twitter Users.

BACKGROUND: The COVID-19 pandemic prompted global behavioral restrictions, impacting public mental health. Sentiment analysis, a tool for assessing individual and public emotions from text data, gained importance amid the pandemic. This study focuses on Japan's early public health interventions during COVID-19, utilizing sentiment analysis in infodemiology to gauge public sentiment on social media regarding these interventions. OBJECTIVE: This study aims to investigate shifts in public emotions and sentiments before and after the first state of emergency was declared in Japan. By analyzing both user-generated tweets and retweets, we aim to discern patterns in emotional responses during this critical period. METHODS: We conducted a day-by-day analysis of Twitter (now known as X) data using 4,894,009 tweets containing the keywords "corona," "COVID-19," and "new pneumonia" from March 23 to April 21, 2020, approximately 2 weeks before and after the first declaration of a state of emergency in Japan. We also processed tweet data into vectors for each word, employing the Fuzzy-C-Means (FCM) method, a type of cluster analysis, for the words in the sentiment dictionary. We set up 7 sentiment clusters (negative: anger, sadness, surprise, disgust; neutral: anxiety; positive: trust and joy) and conducted sentiment analysis of the tweet groups and retweet groups. RESULTS: The analysis revealed a mix of positive and negative sentiments, with "joy" significantly increasing in the retweet group after the state of emergency declaration. Negative emotions, such as "worry" and "disgust," were prevalent in both tweet and retweet groups. Furthermore, the retweet group had a tendency to share more negative content compared to the tweet group. CONCLUSIONS: This study conducted sentiment analysis of Japanese tweets and retweets to explore public sentiments during the early stages of COVID-19 in Japan, spanning 2 weeks before and after the first state of emergency declaration. The analysis revealed a mix of positive (joy) and negative (anxiety, disgust) emotions. Notably, joy increased in the retweet group after the emergency declaration, but this group also tended to share more negative content than the tweet group. This study suggests that the state of emergency heightened positive sentiments due to expectations for infection prevention measures, yet negative information also gained traction. The findings propose the potential for further exploration through network analysis.

Scientific techniques in adolescent and young adult classic Hodgkin lymphoma.

Understanding the tumor microenvironment and genomic landscape is crucial for better prediction of treatment outcomes and developing novel therapies in Hodgkin lymphoma (HL). Recent advancements in genomics have enabled researchers to gain deeper insights into the genomic characteristics of HL at both single-cell resolution and the whole genome level. The use of noninvasive methods such as liquid biopsies and formalin-fixed paraffin-embedded-based imaging techniques has expanded the possibilities of applying cutting-edge analyses to routine clinically available samples. Collaborative efforts between adult and pediatric group are imperative to translate novel findings into routine patient care.

[Inflammatory Responses in Hemorrhagic Stroke].

Inflammation is triggered by various intrinsic and extrinsic stimuli as a protective machinery to maintain homeostasis in the human body. Usually, it is magnified in intensity initially and regresses rapidly afterwards; this phenomenon is called acute inflammation. However, it occasionally lasts a long time; this phenomenon is called chronic inflammation. Induction of some specific machineries, i.e., formation of a positive feedback loop, inflammatory cell infiltration, and changes in tissue architecture, is required for the transition to chronic inflammation; this differentiates chronic and acute inflammation in nature. Chronic inflammation is a common pathogenesis of various diseases, including cancer, vascular disease, and stroke. Recent experimental studies have clarified the crucial role of inflammatory responses in the development and progression of hemorrhagic stroke mediated by tissue destruction or some other aspects of diseases. In this review, we summarize the research findings of the role of inflammation in hemorrhagic stroke.

Clinicopathological characteristics associated with the engraftment of patient lymphoma cells in NOG mice.

Patient-derived xenograft (PDX) mouse models are useful for deepening our understanding of the biology of malignant lymphoma; however, factors associated with the success of the PDX lymphoma model are largely unknown. We retrospectively analyzed the characteristics of 66 xenotransplantations from 65 patients. In all, 43 (65%) specimens were obtained from patients aged > 60 years, and 42 (64%) specimens were obtained at diagnosis. Specimens were obtained from patients with the following diseases: diffuse large B-cell lymphoma (n = 30), intravascular large B-cell lymphoma (n = 12), follicular lymphoma (n = 8), peripheral T-cell lymphoma (n = 7), mantle cell lymphoma (n = 2), and other (n = 7). The specimens were sourced mainly from bone marrow (n = 31, 47%) and extranodal tumors (n = 13, 20%). Engraftment was successful in 33/66 (50%) xenotransplantations. The median age of patients who provided successful specimens was significantly higher than that for unsuccessful specimens (p = 0.013). Specimens with a high proportion of tumor cells in the graft and those obtained from patients with relapsed/refractory disease showed higher tendencies toward successful engraftment. Taken together, these data suggest that tumor cells with a highly malignant potential might have a high likelihood of engraftment.

Hypoxic microenvironment as a crucial factor triggering events leading to rupture of intracranial aneurysm.

Subarachnoid hemorrhage being the rupture of intracranial aneurysm (IA) as a major cause has quite poor prognosis, despite the modern technical advances. Thereby, the mechanisms underlying the rupture of lesions should be clarified. Recently, we and others have clarified the formation of vasa vasorum in IA lesions presumably for inflammatory cells to infiltrate in lesions as the potential histopathological alternation leading to rupture. In the present study, we clarified the origin of vasa vasorum as arteries located at the brain surface using 3D-immunohistochemistry with tissue transparency. Using Hypoxyprobe, we then found the presence of hypoxic microenvironment mainly at the adventitia of intracranial arteries where IA is formed. In addition, the production of vascular endothelial growth factor (VEGF) from cultured macrophages in such a hypoxic condition was identified. Furthermore, we found the accumulation of VEGF both in rupture-prone IA lesions induced in a rat model and human unruptured IA lesions. Finally, the VEGF-dependent induction of neovessels from arteries on brain surface was confirmed. The findings from the present study have revealed the potential role of hypoxic microenvironment and hypoxia-induced VEGF production as a machinery triggering rupture of IAs via providing root for inflammatory cells in lesions to exacerbate inflammation.

Novel insights into Hodgkin lymphoma biology by single-cell analysis.

The emergence and rapid development of single-cell technologies mark a paradigm shift in cancer research. Various technology implementations represent powerful tools to understand cellular heterogeneity, identify minor cell populations that were previously hard to detect and define, and make inferences about cell-to-cell interactions at single-cell resolution. Applied to lymphoma, recent advances in single-cell RNA sequencing have broadened opportunities to delineate previously underappreciated heterogeneity of malignant cell differentiation states and presumed cell of origin, and to describe the composition and cellular subsets in the ecosystem of the tumor microenvironment (TME). Clinical deployment of an expanding armamentarium of immunotherapy options that rely on targets and immune cell interactions in the TME emphasizes the requirement for a deeper understanding of immune biology in lymphoma. In particular, classic Hodgkin lymphoma (CHL) can serve as a study paradigm because of its unique TME, featuring infrequent tumor cells among numerous nonmalignant immune cells with significant interpatient and intrapatient variability. Synergistic to advances in single-cell sequencing, multiplexed imaging techniques have added a new dimension to describing cellular cross talk in various lymphoma entities. Here, we comprehensively review recent progress using novel single-cell technologies with an emphasis on the TME biology of CHL as an application field. The described technologies, which are applicable to peripheral blood, fresh tissues, and formalin-fixed samples, hold the promise to accelerate biomarker discovery for novel immunotherapeutic approaches and to serve as future assay platforms for biomarker-informed treatment selection, including immunotherapies.

Associations Between Drug Treatments and the Risk of Aneurysmal Subarachnoid Hemorrhage: a Systematic Review and Meta-analysis.

There is increasing interest in drug therapy for preventing aneurysmal subarachnoid hemorrhage (aSAH). We aimed to comprehensively evaluate the association between drug use and the risk of aSAH. We searched PubMed and Scopus from the databases' inception until December 2021. Observational studies reporting the association between any drug therapy and aSAH were included. The odds ratios (ORs) for each drug used in aSAH were meta-analyzed with a random-effect model. According to the systematic review, 25 observational studies were eligible for the present study. Four therapeutic purpose-based classes (e.g., lipid-lowering agents) and 14 mechanism-based classes (e.g., statins) were meta-analyzed. Anti-hypertensive agents (OR, 0.50; 95% confidence interval [95% CI], 0.33-0.74), statins (OR, 0.55; 95% CI, 0.35-0.85), biguanides (OR, 0.57; 95% CI, 0.34-0.96), and acetylsalicylic acid (ASA) (OR, 0.62; 95% CI, 0.41-0.94) were inversely associated with the risk of aSAH. Non-ASA non-steroidal anti-inflammatory drugs (OR, 1.73; 95% CI, 1.07-2.79), selective cyclooxygenase-2 inhibitors (OR, 2.04; 95% CI, 1.24-3.35), vitamin K antagonists (OR, 1.50; 95% CI, 1.18-1.91), and dipyridamole (OR, 1.77; 95% CI, 1.23-2.54) were positively associated with the incidence of aSAH. There was also a trend toward a positive association between glucocorticoids (OR, 1.38; 95% CI, 0.97-1.94) and aSAH. The present study suggests that anti-hypertensive agents, statins, biguanides, and ASA are candidate drugs for preventing aSAH. By contrast, several drugs (e.g., anti-thrombotic drugs) may increase the risk of aSAH. Thus, the indications of these drugs in patients with intracranial aneurysms should be carefully determined.

Endothelial cell malfunction in unruptured intracranial aneurysm lesions revealed using a 3D-casted mold.

Intracranial aneurysms (IA) are major causes of devastating subarachnoid hemorrhages. They are characterized by a chronic inflammatory process in the intracranial arterial walls triggered and modified by hemodynamic force loading. Because IA lesion morphology is complex, the blood flow conditions loaded on endothelial cells in each portion of the lesion in situ vary greatly. We created a 3D-casted mold of the human unruptured IA lesion and cultured endothelial cells on this model; it was then perfused with culture media to model physiological flow conditions. Gene expression profiles of endothelial cells in each part of the IA lesion were then analyzed. Comprehensive gene expression profile analysis revealed similar gene expression patterns in endothelial cells from each part of the IA lesion but gene ontology analysis revealed endothelial cell malfunction within the IA lesion. Histopathological examination, electron microscopy, and immunohistochemical analysis indicated that endothelial cells within IA lesions are damaged and dysfunctional. Thus, our findings reveal endothelial cell malfunction in IA lesions and provided new insights into IA pathogenesis.

Identification of The Unique Subtype of Macrophages in Aneurysm Lesions at the Growth Phase.

OBJECTIVES: Recent experimental studies have defined intracranial aneurysms as a macrophage-mediated chronic inflammatory disease affecting intracranial arteries. Although there are various subtypes in macrophages, what type of macrophages is present in lesions during the disease development remains to be elucidated. METHODS: The previously-established aneurysm model of rats was used. Macrophages were labeled with the fluorescent protein and isolated by a laser-microdissection method. The comprehensive gene expression profile analyses and gene ontology analyses was then done to identify a macrophage subtype present in lesions at the growth phase. RESULTS: The gene expression profile data of total 52 macrophages infiltrating into the lesions was acquired. The principal component analysis revealed the monotonous macrophage subtype. By comparing the profile identified with one from in vitro-differentiated M0 or M1 macrophages, the macrophages in the lesions were belonged to the simple and unique subtype. Because the perception of signaling from nervous system was highlighted as up-represented terms through gene ontology analyses, the macrophage subtype in lesions at the growth phase might be differentiated under the influence of nervous system in the microenvironment. The histopathological examinations supported the above notion by confirming the presence of nerves in the adventitia. CONCLUSIONS: The findings from the present study have provided the useful insights about the macrophage subtype in aneurysm lesions at the growth phase and also proposed its ability as a therapeutic target.

Single-cell profiling reveals a memory B cell-like subtype of follicular lymphoma with increased transformation risk.

Follicular lymphoma (FL) is an indolent cancer of mature B-cells but with ongoing risk of transformation to more aggressive histology over time. Recurrent mutations associated with transformation have been identified; however, prognostic features that can be discerned at diagnosis could be clinically useful. We present here comprehensive profiling of both tumor and immune compartments in 155 diagnostic FL biopsies at single-cell resolution by mass cytometry. This revealed a diversity of phenotypes but included two recurrent patterns, one which closely resembles germinal center B-cells (GCB) and another which appears more related to memory B-cells (MB). GCB-type tumors are enriched for EZH2, TNFRSF14, and MEF2B mutations, while MB-type tumors contain increased follicular helper T-cells. MB-type and intratumoral phenotypic diversity are independently associated with increased risk of transformation, supporting biological relevance of these features. Notably, a reduced 26-marker panel retains sufficient information to allow phenotypic profiling of future cohorts by conventional flow cytometry.

ERRATUM: Differences in spatial patterns of long-term care depending on severity in Hokkaido, Japan.

Due to a mistake, the authors' affiliations were incorrectly reported in this article, published in Geospatial Health in 2022 (DOI: 10.4081/gh.2022.1077 - PMID: 35579241). The correct affiliations appear above. Geospatial Health DOI: 10.4081/gh.2022.1137.

Potential crowdedness of mechanical thrombectomy and cerebral infarction mortality in Japan: Application of inverted two-step floating catchment area method.

OBJECTIVES: This study aimed to evaluate a stroke medical delivery system based on population coverage and the potential crowdedness index (PCI) of mechanical thrombectomy and investigate the relationship between PCI and cerebral infarction mortality in Japan. MATERIALS AND METHODS: This cross-sectional study defined 662 facilities and 1605 neurointerventionalists as supply, population aged 55 years or older as demand, and set the reachable area for demand as 120 min in driving time. Multiple regression analysis adjusted for spatial autocorrelation was used to examine the relationship between PCI and cerebral infarction mortality. RESULTS: In the 2020 data, 99% of the population aged 55 years or older had access to mechanical thrombectomy (≤120 min), and the PCI ranged from 5876 to 129838, with a median of 30426. From 2020 to 2035, the PCI is estimated to increase (30426 to 32510), decreasing after 2035 (32510 to 29469). The PCI distribution exhibited geographical heterogeneity. High PCI values emerged in eastern Japan. According to regression analysis, the increase in PCI by 1% led to an increase of 0.13% in standardized mortality ratio of cerebral infarction in men. However, PCI did not significantly correlate with cerebral infarction mortality in women. CONCLUSIONS: PCI for hospitals based on supply and demand was geographically heterogeneous in Japan. Optimization of PCI contributes equalization of mechanical thrombectomy provision system and may improve cerebral infarction mortality.

Single-cell spatial analysis of tumor immune architecture in diffuse large B-cell lymphoma.

Multiplexed immune cell profiling of the tumor microenvironment (TME) in cancer has improved our understanding of cancer immunology, but complex spatial analyses of tumor-immune interactions in lymphoma are lacking. Here, we used imaging mass cytometry (IMC) on 33 cases of diffuse large B-cell lymphoma (DLBCL) to characterize tumor and immune cell architecture and correlate it to clinicopathological features such as cell of origin, gene mutations, and responsiveness to chemotherapy. To understand the poor response of DLBCL to immune checkpoint inhibitors (ICI), we compared our results to IMC data from Hodgkin lymphoma, a cancer highly responsive to ICI, and observed differences in the expression of PD-L1, PD-1, and TIM-3. We created a spatial classification of tumor cells and identified tumor-centric subregions of immune activation, immune suppression, and immune exclusion within the topology of DLBCL. Finally, the spatial analysis allowed us to identify markers such as CXCR3, which are associated with penetration of immune cells into immune desert regions, with important implications for engineered cellular therapies. This is the first study to integrate tumor mutational profiling, cell of origin classification, and multiplexed immuno-phenotyping of the TME into a spatial analysis of DLBCL at the single-cell level. We demonstrate that, far from being histopathologically monotonous, DLBCL has a complex tumor architecture, and that changes in tumor topology can be correlated with clinically relevant features. This analysis identifies candidate biomarkers and therapeutic targets such as TIM-3, CCR4, and CXCR3 that are relevant for combination treatment strategies in immuno-oncology and cellular therapies.

The bifurcation angle is associated with the progression of saccular aneurysms.

The role of the bifurcation angle in progression of saccular intracranial aneurysms (sIAs) has been undetermined. We, therefore, assessed the association of bifurcation angles with aneurysm progression using a bifurcation-type aneurysm model in rats and anterior communicating artery aneurysms in a multicenter case-control study. Aneurysm progression was defined as growth by ≥ 1 mm or rupture during observation, and controls as progression-free for 30 days in rats and ≥ 36 months in humans. In the rat model, baseline bifurcation angles were significantly wider in progressive aneurysms than in stable ones. In the case-control study, 27 and 65 patients were enrolled in the progression and control groups. Inter-observer agreement for the presence or absence of the growth was excellent (κ coefficient, 0.82; 95% CI, 0.61-1.0). Multivariate logistic regression analysis showed that wider baseline bifurcation angles were significantly associated with subsequent progressions. The odds ratio for the progression of the second (145°-179°) or third (180°-274°) tertiles compared to the first tertile (46°-143°) were 5.5 (95% CI, 1.3-35). Besides, the bifurcation angle was positively correlated with the size of aneurysms (Spearman's rho, 0.39; P = 0.00014). The present study suggests the usefulness of the bifurcation angle for predicting the progression of sIAs.

Differences in spatial patterns of long-term care depending on severity in Hokkaido, Japan.

The increasing demand for long-term care (LTC) among the ageing population is a serious problem worldwide, which has greatly increased also in Japan since the introduction of the LTC insurance system there. Since there is a difference between insurers with respect to the proportion of people needing LTC, this study aimed at clarifying the spatial patterns of LTC. Insurer (n=156) LTC data for the period 2012-2019 were obtained from Ministry of Health, Labour, and Welfare and those needing LTC were classified into three classes: total, mild and severe with ageand sex-adjusted proportions needing LTC. Global and local Moran’s I statistics were calculated for each 2-year period to clarify the trends of global and local spatial clusters. From 2012 to 2019, the mean proportion of mild class cases increased (10.6% to 11.6%), whereas that of severe class cases decreased slightly (5.9% to 5.7%). The spatial pattern of the proportion of each class revealed positive spatial autocorrelation. Based on analysis by local Moran’s I, differences in spatial patterns were emphasised between the mild and severe classes. In Hokkaido, High-High clusters of mild cases were identified in the central and southern parts and severe ones in the northern and southern parts. Spatial patterns differed depending on the LTC class. Some insurers had distinctly higher or lower certification rates than those of their neighbourhoods.