Detection of Novel Tyrosine Kinase Fusion Genes as Potential Therapeutic Targets in Bone and Soft Tissue Sarcomas Using DNA/RNA-based Clinical Sequencing.

BACKGROUND: Approximately 1% of clinically treatable tyrosine kinase fusions, including anaplastic lymphoma kinase, neurotrophic tyrosine receptor kinase, RET proto-oncogene, and ROS proto-oncogene 1, have been identified in soft tissue sarcomas via comprehensive genome profiling based on DNA sequencing. Histologic tumor-specific fusion genes have been reported in approximately 20% of soft tissue sarcomas; however, unlike tyrosine kinase fusion genes, these fusions cannot be directly targeted in therapy. Approximately 80% of tumor-specific fusion-negative sarcomas, including myxofibrosarcoma and leiomyosarcoma, that are defined in complex karyotype sarcomas remain genetically uncharacterized; this mutually exclusive pattern of mutations suggests that other mutually exclusive driver oncogenes are yet to be discovered. Tumor-specific, fusion-negative sarcomas may be associated with unique translocations, and oncogenic fusion genes, including tyrosine kinase fusions, may have been overlooked in these sarcomas. QUESTIONS/PURPOSES: (1) Can DNA- or RNA-based analysis reveal any characteristic gene alterations in bone and soft tissue sarcomas? (2) Can useful and potential tyrosine kinase fusions in tumors from tumor-specific, fusion-negative sarcomas be detected using an RNA-based screening system? (3) Do the identified potential fusion tumors, especially in neurotrophic tyrosine receptor kinase gene fusions in bone sarcoma, transform cells and respond to targeted drug treatment in in vitro assays? (4) Can the identified tyrosine kinase fusion genes in sarcomas be useful therapeutic targets? METHODS: Between 2017 and 2020, we treated 100 patients for bone and soft tissue sarcomas at five institutions. Any biopsy or surgery from which a specimen could be obtained was included as potentially eligible. Ninety percent (90 patients) of patients were eligible; a further 8% (8 patients) were excluded because they were either lost to follow-up or their diagnosis was changed, leaving 82% (82 patients) for analysis here. To answer our first and second questions regarding gene alterations and potential tyrosine kinase fusions in eight bone and 74 soft tissue sarcomas, we used the TruSight Tumor 170 assay to detect mutations, copy number variations, and gene fusions in the samples. To answer our third question, we performed functional analyses involving in vitro assays to determine whether the identified tyrosine kinase fusions were associated with oncogenic abilities and drug responses. Finally, to determine usefulness as therapeutic targets, two pediatric patients harboring an NTRK fusion and an ALK fusion were treated with tyrosine kinase inhibitors in clinical trials. RESULTS: DNA/RNA-based analysis demonstrated characteristic alterations in bone and soft tissue sarcomas; DNA-based analyses detected TP53 and copy number alterations of MDM2 and CDK4 . These single-nucleotide variants and copy number variations were enriched in specific fusion-negative sarcomas. RNA-based screening detected fusion genes in 24% (20 of 82) of patients. Useful potential fusions were detected in 19% (11 of 58) of tumor-specific fusion-negative sarcomas, with nine of these patients harboring tyrosine kinase fusion genes; five of these patients had in-frame tyrosine kinase fusion genes ( STRN3-NTRK3, VWC2-EGFR, ICK-KDR, FOXP2-MET , and CEP290-MET ) with unknown pathologic significance. The functional analysis revealed that STRN3-NTRK3 rearrangement that was identified in bone had a strong transforming potential in 3T3 cells, and that STRN3-NTRK3 -positive cells were sensitive to larotrectinib in vitro. To confirm the usefulness of identified tyrosine kinase fusion genes as therapeutic targets, patients with well-characterized LMNA-NTRK1 and CLTC-ALK fusions were treated with tyrosine kinase inhibitors in clinical trials, and a complete response was achieved. CONCLUSION: We identified useful potential therapeutic targets for tyrosine kinase fusions in bone and soft tissue sarcomas using RNA-based analysis. We successfully identified STRN3-NTRK3 fusion in a patient with leiomyosarcoma of bone and determined the malignant potential of this fusion gene via functional analyses and drug effects. In light of these discoveries, comprehensive genome profiling should be considered even if the sarcoma is a bone sarcoma. There seem to be some limitations regarding current DNA-based comprehensive genome profiling tests, and it is important to use RNA testing for proper diagnosis and accurate identification of fusion genes. Studies on more patients, validation of results, and further functional analysis of unknown tyrosine kinase fusion genes are required to establish future treatments. CLINICAL RELEVANCE: DNA- and RNA-based screening systems may be useful for detecting tyrosine kinase fusion genes in specific fusion-negative sarcomas and identifying key therapeutic targets, leading to possible breakthroughs in the treatment of bone and soft tissue sarcomas. Given that current DNA sequencing misses fusion genes, RNA-based screening systems should be widely considered as a worldwide test for sarcoma. If standard treatments such as chemotherapy are not effective, or even if the sarcoma is of bone, RNA sequencing should be considered to identify as many therapeutic targets as possible.

Expression of paired box 9 defines an aggressive subset of lung adenocarcinoma preferentially occurring in smokers.

AIMS: A distinct subset of lung adenocarcinomas (LADs), arising from a series of peripheral lung cells defined as the terminal respiratory unit (TRU), is characterised by thyroid transcription factor 1 (TTF-1) expression. The clinical relevance of transcription factors (TFs) other than TTF-1 remains unknown in LAD and was explored in the present study. METHODS AND RESULTS: Seventy-one LAD samples were subjected to high-throughput transcriptome screening of LAD using cap analysis gene expression (CAGE) sequencing data; CAGE provides genome-wide expression levels of the transcription start sites (TSSs). In total, 1083 invasive LAD samples were subjected to immunohistochemical examination for paired box 9 (PAX9) and TTF-1 expression levels. PAX9 is an endoderm development-associated TF that most strongly and inversely correlates with the expression of TTF-1 TSS subsets. Immunohistochemically, PAX9 expression was restricted to the nuclei of ciliated epithelial and basal cells in the bronchi and bronchioles and the nuclei of epithelial cells of the bronchial glands; moreover, PAX9 expression was observed in 304 LADs (28%). PAX9-positive LADs were significantly associated with heavy smoking, non-lepidic subtype, EGFR wild-type tumours and PD-L1 expression (all P < 0.0001). All these characteristics were opposite to those of TRU-type LADs with TTF-1 expression. PAX9 expression was an independent prognostic factor for decreased overall survival (P = 0.022). CONCLUSIONS: Our results revealed that PAX9 expression defines an aggressive subset of LADs preferentially occurring in smokers that may arise from bronchial or bronchiolar cells.

Effects of social isolation associated with the COVID-19 pandemic on hip muscle strength, hip joint pain, and walking ability in patients with osteoarthritis of the hip joint.

To reduce the spread of the Sars-CoV-2 virus, governments in many countries adopted a social isolation strategy. However, social isolation may adversely affect people's health, e.g., by decreasing the muscle function of lower limbs. We recruited 118 patients who had undergone total hip arthroplasty (THA) and 87 patients with moderate to severe hip joint osteoarthritis (OA) and measured hip muscle strength, hip joint pain, and walking ability from before to one year after the start of the COVID-19 pandemic. During the pandemic, hip flexion (straight leg raise, SLR) strength decreased in 13.1% of patients in the post-THA group and 25.6% in the severe-OA group; in the severe-OA group, the decrease in SLR strength was mainly in patients aged 65 years and older. In addition, pain increased to mild or moderate and walkable distance decreased in more patients in the severe-OA group.

Diffuse expression of MUC6 defines a distinct clinicopathological subset of pulmonary invasive mucinous adenocarcinoma.

Invasive mucinous adenocarcinoma (IMA) of the lung is a unique variant of lung adenocarcinoma. Aberrant mucin expression is associated with cancer development and metastasis. However, the clinicopathological significance of mucin expression in IMA is not fully understood. Herein, we evaluated the clinicopathological, immunohistochemical, and molecular characteristics of 70 IMA tumors. EGFR, KRAS, GNAS, and TP53 mutations were assessed by PCR-based sequencing. Next-generation sequencing was used to assess cases without EGFR/KRAS mutations. A NanoString-based screening for fusions was performed in all IMAs without mitogenic driver mutations. Expression of mucins (MUC1, MUC2, MUC4, MUC5AC, and MUC6) was evaluated by immunohistochemistry and categorized as follows: negative (<10% of tumor cells), patchy expression (<90% of tumor cells), or diffuse expression (≥90% of tumor cells). Immunohistochemical testing for transcription factors (TTF-1, CDX2, HNF1ß, HNF3α, HNF3ß, and HNF4α) was also performed. As expected, KRAS mutations were the most common (in 67% of cases), followed by small numbers of other alterations. Patchy or diffuse expression of MUC1, MUC2, MUC4, MUC5AC, and MUC6 was observed in 52% or 6%, 3% or 0%, 30% or 3%, 26% or 73%, and 59% or 27% of cases, respectively. Furthermore, all IMAs were generally positive for HNF1ß (100%), HNF3α (100%), HNF3ß (100%), and HNF4α (99%) but were positive less often for TTF-1 (6%) and CDX2 (9%). Overall, there was no significant correlation between mucin expression and transcription factor expression. Unexpectedly, diffuse expression of MUC6 was significantly associated with KRAS-wild-type tumors (p = 0.0008), smaller tumor size (p = 0.0073), and tumors in female patients (p = 0.0359) in multivariate analyses. Furthermore, patients with tumors exhibiting diffuse MUC6 expression had significantly favorable outcomes. Notably, none of these patients died of the disease. Our data suggested that diffuse expression of MUC6 defines a distinct clinicopathological subset of IMA characterized by wild-type KRAS and possibly less aggressive clinical course.

Histological characteristics of lung adenocarcinoma with uncommon actionable alterations: special emphasis on MET exon 14 skipping alterations.

AIMS: In the evolving era of precision medicine, increasing emphasis is placed on detecting molecular alterations driving the development of specific cancers and targeting them with matched therapies that can yield the best outcomes for patients. Lung adenocarcinomas with uncommon actionable alterations, including MET exon 14 skipping (METex14), ERBB2 and BRAF mutations, are rare and poorly characterised cancers. METHODS AND RESULTS: To more clearly understand the histopathological features of lung adenocarcinoma with uncommon actionable alterations, we compared the histological features of 678 cases with mitogenic driver alterations from 996 surgically resected lung adenocarcinomas. Genomic data from our cohort revealed METex14, ERBB2 and BRAF mutations in 13, 16 and 15 cases, respectively. Patients who had lung adenocarcinoma with METex14 were often elderly females. Histological features such as clear cell features (23%), hyaline globules (31%) and nuclear pleomorphism (39%) were the most frequently identified in METex14-positive cases; among those, three cases (23%) had tumour cells with bizarre giant or multilobulated nuclei. Additionally, the micropapillary pattern was the most frequently identified in ERBB2-mutated lung adenocarcinoma (31%). Lung adenocarcinoma with BRAF mutations tended to be less invasive, and the BRAF V600E mutation was identified in only one case with lepidic adenocarcinoma. Immunohistochemically, all METex14, ERBB2 and BRAF-positive tumours, except for invasive mucinous adenocarcinoma, were positive for thyroid transcription factor 1 (TTF-1). CONCLUSIONS: Our data from Japanese patients showed that lung adenocarcinoma with METex14 had unique clinicopathological characteristics: tumour cells with marked nuclear pleomorphism, hyaline globules and expression of TTF-1 in elderly women who never or lightly smoked.

Identification of a Novel MAN1A1-ROS1 Fusion Gene Through mRNA-based Screening for Tyrosine Kinase Gene Aberrations in a Patient with Leiomyosarcoma.

BACKGROUND: Soft tissue sarcomas are a heterogeneous group of rare malignant tumors. Advanced soft tissue sarcomas have a poor prognosis, and effective systemic therapies have not been established. Tyrosine kinases are increasingly being used as therapeutic targets for a variety of cancers and soft tissue sarcomas. Although complex karyotype sarcomas typically tend to carry more potentially actionable genetic alterations than do translocation-associated sarcomas (fusion gene sarcomas), based on our database review, we found that leiomyosarcoma and malignant peripheral nerve sheath tumors have lower frequencies of potential targets than other nontranslocation soft tissue sarcomas. We theorized that both leiomyosarcoma and malignant peripheral nerve sheath tumors might be included in any unique translocations. Furthermore, if tyrosine kinase imbalances, especially fusion genes, occur in patients with leiomyosarcomas and malignant peripheral nerve sheath tumors, tyrosine kinase inhibitors might be a drug development target for this sarcoma. In this study, we used a tyrosine kinase screening system that could detect an imbalance in mRNA between 5'- and 3'-sides in tyrosine kinase genes to identify potential novel therapeutic tyrosine kinase targets for soft tissue sarcomas. QUESTIONS/PURPOSES: (1) Are there novel therapeutic tyrosine kinase targets in tumors from patients with soft tissue sarcomas that are detectable using mRNA screening focusing on imbalance expressions between the 5' and 3' end of the kinase domain? (2) Can potential targets be verified by RNA sequencing and reverse transcription PCR (RT-PCR)? (3) Will potential fusion gene(s) transform cells in in vitro assays? (4) Will tumors in mice that have an identified fusion gene respond to treatment with a therapeutic drug directed at that target? METHODS: We used mRNA screening to look for novel tyrosine kinase targets that might be of therapeutic potential. Using functional assays, we verified whether the identified fusion genes would be good therapeutic candidates for soft tissue sarcomas. Additionally, using in vivo assays, we assessed whether suppressing the fusion's kinase activity has therapeutic potential. Study eligibility was based on a patient having high-grade spindle cell and nontranslocation sarcomas, including leiomyosarcoma, malignant peripheral nerve sheath tumor, and high-grade myxofibrosarcoma. Between 2015 and 2019, of the 172 patients with soft tissue sarcomas treated with surgical resection at Juntendo University Hospital, 72 patients had high-grade nontranslocation sarcomas. The analysis was primarily for leiomyosarcoma and malignant peripheral nerve sheath tumors, and there was a limitation of analysis size (reagent limitations) totaling 24 samples at the start of the study. We collected additional samples from a sample bank at the Tokyo Medical and Dental University to increase the number of sarcomas to study. Therefore, in this study, a total of 15 leiomyosarcoma samples, five malignant peripheral nerve sheath tumors samples, and four high-grade myxofibrosarcoma samples were collected to achieve the sample size of 24 patients. To identify tyrosine kinase fusion genes, we designed a NanoString-based assay (NanoString Technologies Inc, Seattle, WA, USA) to query the expression balances regarding transcripts of 90 tyrosine kinases at two points: the 5' end of the kinase domain and within the kinase domain or 3' end of the kinase domain. The tumor's RNA was hybridized to the NanoString probes and analyzed for the expression ratios of outliers from the 3' to 5' end of the kinase domain. Presumed novel fusion events in these positive tumors that were defined by NanoString-based assays were confirmed tyrosine kinase fusion genes by RNA sequencing and confirmatory RT-PCR. Functional analyses consisting of in vitro and in vivo assays were also performed to elucidate whether the identified tyrosine kinase gene fusions were associated with oncogenic abilities and drug responses. RESULTS: We identified aberrant expression ratios regarding the 3' to 5' end of the kinase domain ratios in ROS1 transcripts in a leiomyosarcoma in a 90-year-old woman. A novel MAN1A1-ROS1 fusion gene was identified from her thigh tumor through RNA sequencing, which was confirmed with real-time PCR. In functional assays, MAN1A1-ROS1 rearrangement revealed strong transforming potential in 3T3 cells. Moreover, in an in vivo assay, crizotinib, a ROS1 inhibitor, markedly inhibited the growth of MAN1A1-ROS1 rearrangement-induced transformed cells in a dose-dependent manner. CONCLUSION: We conducted tyrosine kinase screening to identify new therapeutic targets in soft tissue sarcomas. We found a novel MAN1A1-ROS1 fusion gene that may be a therapeutic target in patients with leiomyosarcoma. This study demonstrates that the mRNA screening system may aid in the development of useful therapeutic options for soft tissue sarcomas. CLINICAL RELEVANCE: If novel tyrosine fusions such as MAN1A1-ROS1 fusion can be found in sarcomas from other patients, they could offer avenues for new molecular target therapies for sarcomas that currently do not have effective chemotherapeutic options. Therefore, the establishment of a screening system that includes both genomic and transcript analyses in the clinical setting is needed to verify our discoveries and take the developmental process of treatment to the next step.

Assessment of Predictive Biomarkers of the Response to Pazopanib Based on an Integrative Analysis of High-grade Soft-tissue Sarcomas: Analysis of a Tumor Sample from a Responder and Patients with Other Soft-tissue Sarcomas.

BACKGROUND: Soft-tissue sarcomas are a rare group of malignant tumors that usually are treated with surgical excision and radiation therapy, but recently, pazopanib, an oral tyrosine kinase inhibitor, has been used in patients with metastases who do not respond to standard chemotherapy regimens. Based on patients with advanced soft-tissue sarcomas who had received prior chemotherapy, several clinical studies have reported the survival and sensitivity (approximately 5% to 10% sensitive) of patients with soft-tissue sarcomas treated with pazopanib. Recently, next-generation sequencing (NGS) technologies have been used to provide a wide genetic information and to develop personalized medicine in cancer treatment. However, there are few reports and no genetic analyses of patients with soft-tissue sarcomas who had a complete response (CR) to pazopanib. QUESTIONS/PURPOSES: We described the clinicopathologic features of a patient with a rare, advanced soft-tissue sarcoma who achieved a CR to pazopanib treatment. Furthermore, integrative analyses using NGS and arrays were performed to elucidate characteristic alterations, including gene mutations, copy number changes, and protein expression that were associated with response to pazopanib. Additionally, functional analyses consisting of in vitro and in vivo assays were also performed to elucidate whether the identified alterations were associated with oncogenic abilities and drug responses. METHODS: In a sample from a 70-year-old woman with an advanced soft-tissue sarcoma treated for 1 month with 800 mg of oral pazopanib daily, CT scans demonstrated a CR to treatment. To our knowledge, there have been no patients with soft-tissue sarcomas among several clinical trials of pazopanib that have achieved a CR and therefore, our patient is considered to be extremely rare. We performed an integrative analysis including whole-exome sequencing, transcriptome sequencing, and phosphorylation profiling of receptor tyrosine kinases (RTK) using tumor samples from a patient with a CR matched to normal samples. From here on we will refer to this patient as having a CR, although a short term high-grade partial response may be more accurate. These analyses were performed using NGS and the phosphoreceptor tyrosine kinase (phospho-RTK) array. As a validation study, we also performed target sequencing using three samples from patients with long-term stable disease and two samples from patients with progressive disease who responded to pazopanib treatment. In addition, characteristic gene alterations that were identified according to the response to pazopanib in one patient with a CR, in three patients with long-term stable disease, and in 27 patients with high-grade soft-tissue sarcomas with different histologic subtypes and different responses to pazopanib were verified by quantitative real-time polymerase chain reaction. We conducted a focus formation assay to evaluate the transforming activities of these genomic alterations. RESULTS: In the patient with a CR to pazopanib, we identified several somatic mutations including Fms related receptor tyrosine kinase 1 (FLT1) p.G38S, platelet-derived growth factor receptor alpha (PDGFRA) p.T83S, and platelet-derived growth factor receptor beta (PDGFRB) exon 13 skipping. Amplification at chromosome 12q13-14 encompassing GLI family zinc finger 1 (GLI1) and cyclin-dependent kinase-4 (CDK4) was also detected. Furthermore, an elevated PDGFRB phosphorylation level was observed in the tumor. In target sequencing analyses in five patients, one of three patients with long-term stable disease had 12q13-14 amplification. The mRNA expression of GLI1, CDK4, and pazopanib targets including PDGFRA, PDGFRB, vascular endothelial growth factor receptor (VEGFR)1-3, and stem cell factor receptor (KIT) in samples from the patient with a CR, and 27 patients with high-grade soft-tissue sarcomas was verified. The expression of GLI1 was characteristically increased in the patient with a CR and in those with long-term stable disease relative to other patients with soft-tissue sarcomas. Overexpression of GLI1 showed strong transforming potential in 3T3 cells. Moreover, the overexpression of GLI1 upregulated the expression of the PDGFRB protein and promoted phosphorylation, which was dose-dependently inhibited by pazopanib. However, inhibition of GLI1-induced transformation by pazopanib was limited in the focus formation assay; therefore, mechanisms other than PDGFRB activation may contribute to transformation. CONCLUSIONS: We identified several gene alterations that might be associated with a CR and long-term stable disease in patients who received pazopanib for advanced soft-tissue sarcomas. We therefore believe that this distinct molecular profile warrants further investigation to identify predictive biomarkers of the response to pazopanib. CLINICAL RELEVANCE: Our findings identify molecular mechanisms that possibly explain the high sensitivity of soft-tissue sarcomas to pazopanib and may lead to the development of predictive biomarkers and novel therapies in patients with this and other types of soft-tissue sarcomas.

Lateral femoral cutaneous nerve injury with the direct anterior approach for total hip arthroplasty.

PURPOSE: Although injury of the lateral femoral cutaneous nerve (LFCN) is a known complication after total hip arthroplasty (THA) via the direct anterior approach (DAA), the impact of this complication on postoperative quality of life (QOL) is unclear. This study aims to investigate the incidence of LFCN injury after DAA for THA, and to determine the impact of LFCN injury on QOL and hip function. METHODS: We prospectively investigated 122 hips operated upon using the DAA regarding the incidence of LFCN injury using self-reported questionnaires, hip functional scores such as the Harris Hip Score (HHS) and the Japanese Orthopaedic Association (JOA) score, and patient-reported outcomes such as the Western Ontario and McMaster Universities Osteoarthritis Index, the Japanese Orthopaedic Association Hip disease Evaluation Questionnaire (JHEQ), and the Forgotten Joint Score-12 (FJS). RESULTS: LFCN injury was seen in 39 hips (31.9 %). In affected hips, the leading symptom was hypo-aesthesia (46.2 %), followed by tingling or jolt-like sensation (28.2 %). There was no difference in the HHS and JOA score between those with LFCN injury and those without. There was a significant difference in the FJS-12 between the two groups (50.9 ± 25.3 for hips with LFCN injury vs 64.3 ± 25.7 without, p = 0.01). There was a non-significant tendency for patients with LFCN injury to have a lower JHEQ than those without (63.6 ± 19.6 for hips with LFCN injury vs 70.8 ± 22.9 without, p = 0.13). CONCLUSIONS: The incidence of LFCN injury decreased QOL but not hip function after DAA for THA.

Association Between Days of the Week and Intraocular Pressure: Japan Ningen Dock Study.

PRCIS: The analysis of intraocular pressure (IOP) by day of the week using the mega database showed a periodic weekly pattern with the highest value on Monday. PURPOSE: To evaluate IOP by the day of the week. PATIENTS AND METHODS: Annual health checkup examinees between April 2014 and March 2015 were cross-sectionally evaluated. As a result, 655,818 participants [51.5±10.5 (range: 20-96) years, 40.1% women] from 103 medical centers were included. IOP was measured using a noncontact tonometer. The mean IOPs of each day of the week were compared using multiple comparison test and multiple linear regression analysis. Wednesday was set as the reference. Moreover, weekly IOP variations stratified by sex and age were also evaluated. RESULTS: Mean IOPs from Monday to Sunday were 13.19±2.97, 13.06±2.92, 13.05±2.91, 13.05±2.92, 13.12±2.94, 13.10±2.96, and 13.16±2.78 mm Hg. IOP was significantly higher on Monday, Friday, and Saturday than those on Wednesday ( P <0.001, <0.001, 0.002). After adjusting for factors affecting IOP, the IOPs on Monday and Saturday were higher than those on Wednesday [ß=0.097 (95% CI: 0.074-0.121), P <0.001; ß=0.032 (95% CI: 0.005-0.059), P =0.019]. Men had significantly higher IOPs on Monday and Saturday than on Wednesday [ß=0.142 (95% CI: 0.110-0.173), P <0.001; ß=0.053 (95% CI: 0.017-0.089), P =0.004], whereas women did not have a significant trend. Participants aged below 65 years had higher IOPs on Monday ( P <0.001 in under 60 years; P =0.003 in 60-64 years), while those aged 65 years or older did not ( P =0.856). CONCLUSION: IOP values may have a periodic weekly pattern. The high IOP on Monday was more pronounced in men aged less than 65 years.

Comparative Evaluation of Fundus Image Interpretation Accuracy in Glaucoma Screening Among Different Physician Groups.

Purpose: To examine the variability in glaucoma screening using fundus images among physicians, including non-ophthalmologists. Patients and Methods: Sixty-nine eyes from 69 patients, including 25 eyes with glaucoma, were included from the Jikei University Hospital from July 2019 to December 2022. Fundus images were captured using TRC-NW8 (Topcon Corporation, Tokyo, Japan), and were interpreted by 10 non-ophthalmologists, 10 non-specialist ophthalmologists, and 9 specialists for diagnostic accuracy. We analyzed differences in diagnostic accuracy among the three groups. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and Kappa coefficient were compared, using the Kruskal-Wallis test followed by a post hoc Dunn's test. Results: The sensitivity and specificity were 0.22 and 0.92 for non-ophthalmologists, 0.49 and 0.83 for non-specialist ophthalmologists, and 0.68 and 0.87 for specialists, respectively. Both specialists and non-specialist ophthalmologists showed significantly higher sensitivity than non-ophthalmologists (Dunn's test, P<0.001 and P=0.031). There was no significant difference in specificity among the three groups (Kruskal-Wallis test, P=0.086). The PPV did not differ significantly between the groups (Kruskal-Wallis test, P=0.108), while the NPV was significantly higher in specialists compared to non-ophthalmologists (Dunn's test, P<0.001). Specialists also had a significantly higher Kappa coefficient than non-ophthalmologists and non-specialist ophthalmologists (Dunn's test, P<0.001 and P=0.024). Conclusion: Diagnostic accuracy varied significantly based on the physician's background.

Dynamic mode decomposition with memory.

This study proposed a numerical method of dynamic mode decomposition with memory (DMDm) to analyze multidimensional time-series data with memory effects. The memory effect is a widely observed phenomenon in physics and engineering and is considered to be the result of interactions between the system and environment. Dynamic mode decomposition (DMD) is a linear operation-based, data-driven method for multidimensional time-series data proposed in 2008. Although DMD is a successful method for time-series data analysis, it is based on ordinary differential equations and thus cannot incorporate memory effects. In this study, we formulated the abstract algorithmic structure of DMDm and demonstrate its utility in overcoming the memoryless restriction imposed by existing DMD methods on the time-evolution model. In the numerical demonstration, we utilized the Caputo fractional differential to implement an example of DMDm such that the time-series data could be analyzed with power-law memory effects. Thus, we developed a fractional DMD, which is a DMD-based method with arbitrary (real value) order differential operations. The proposed method was applied to synthetic data from a set of fractional oscillators and model parameters were estimated successfully. The proposed method is expected to be useful for scientific applications and aid in model estimation, control, and failure detection of mechanical, thermal, and fluid systems in factory machines, such as modern semiconductor manufacturing equipment.

Associated factors of diabetic retinopathy by artificial intelligence evaluation of fundus images in Japan.

This cross-sectional study aimed to investigate the promoting and inhibitory factors of diabetic retinopathy (DR) according to diabetes mellitus (DM) stage using standardized evaluation of fundus images by artificial intelligence (AI). A total of 30,167 participants underwent blood and fundus examinations at a health screening facility in Japan (2015-2016). Fundus photographs were screened by the AI software, RetCAD and DR scores (DRSs) were quantified. The presence of DR was determined by setting two cut-off values prioritizing sensitivity or specificity. DM was defined as four stages (no DM: DM0; advanced DM: DM3) based on treatment history and hemoglobin A1c (HbA1c) levels. Associated factors of DR were identified using logistic regression analysis. For cutoff values, multivariate analysis revealed age, sex, systolic blood pressure (SBP), smoking, urinary protein, and HbA1c level as positively associated with the risk of DR among all DM stages. In addition to glycemic control, SBP and Fibrosis-4 index might act as promoting factors for DR at all or an earlier DM stage. T-Bil, cholinesterase, and T-cho level might be protective factors at an advanced DM stage.

Lens thickness in atopic cataract: case-control study.

PURPOSE: To examine whether atopic cataracts are associated with thinner lenses. SETTING: Department of Ophthalmology, Jikei University Hospital, Tokyo, Japan. DESIGN: Retrospective matched case-control study. METHODS: 31 eyes with atopic cataracts, 62 with nonatopic cataracts, and 31 without cataracts were analyzed. Each group was matched for age (±4 years) and sex. RESULTS: The mean lens thickness (LT) was 3.76 ± 0.40 mm, 3.94 ± 0.49 mm, and 4.11 ± 0.40 mm in eyes with atopic cataracts, nonatopic cataracts, and normal lenses, respectively. Repeated-measures analysis of variance showed that the LT in the atopic cataract group was significantly thinner than that in the nonatopic cataract ( P = .036) and normal lens ( P < .001) groups. In multivariate logistic regression analysis, a thinner LT was negatively correlated with age (odds ratio [OR], 0.91; 95% CI, 0.86-0.96) and positively correlated with anterior subcapsular cataract (ASC) (OR, 5.61; 95% CI, 1.97-15.99). Atopy was not a significant factor. 24 (38.7%) of the 62 eyes with nonatopic cataracts and 24 (77.4%) of the 31 eyes with atopic cataracts had ASC. CONCLUSIONS: The lenses of eyes with atopic cataracts were thinner than those of controls. Atopic cataracts frequently present with anterior subcapsular opacity, which is associated with lens thinning.

Association Between Alcohol Consumption Patterns and Glaucoma in Japan.

PRCIS: In this case-control study of the Japanese population, including 3207 glaucoma cases, alcohol consumption patterns such as frequency and quantity showed a positive association with glaucoma prevalence. PURPOSE: To examine the association between alcohol consumption patterns and glaucoma. SUBJECTS AND METHODS: This case-control study evaluated 3207 cases with glaucoma and 3207 matched controls. Patients over 40 years of age were included from 1,693,611 patients admitted to 34 hospitals in Japan. Detailed alcohol consumption patterns (drinking frequency, average daily drinks, and total lifetime drinks) were obtained, as well as various confounding factors, including smoking history and lifestyle-related comorbidities. Conditional logistic regression models were used to calculate odds ratios (ORs) and 95% CIs for glaucoma prevalence. RESULTS: Drinking frequency showed an association with glaucoma for "a few days/week" (OR, 1.19; 95% CI, 1.03-1.38) and "almost every day/week" (OR, 1.40; 95% CI, 1.18-1.66). Average daily drinks showed an association for ">0-2 drinks/day" (OR, 1.16; 95% CI, 1.03-1.32). Total lifetime drinks showed an association for ">60-90 drink-year" (OR, 1.23; 95% CI, 1.01-1.49) and ">90 drink-year" (OR, 1.23; 95% CI, 1.05-1.44). As alcohol consumption levels differed considerably between men and women, additional analyses were conducted separately for men and women. Among men, drinking frequency of "a few days/week" and "almost every day/week," average daily drinks of ">0-2 drinks/day" and ">2-4 drinks/day," and total lifetime drinks of ">60-90 drink-year" and ">90 drink-year" had an association with glaucoma. Conversely, among women, neither drinking frequency, average daily drinks, nor total lifetime drinks were associated. CONCLUSIONS: Both the frequency and quantity of alcohol consumption were associated with glaucoma. Further research on gender differences is warranted.

Acetabular morphological variation in Asian patients with femoral neck fracture: A three-dimensional CT-based study.

AIMS: The acetabular morphology varies greatly among individuals, and hypoplasia is more common in Asia than in Europe. Dislocation after bipolar hip arthroplasty (BHA) for femoral neck fracture occurs at a constant rate, and is affected by the acetabular morphology. This study aimed to clarify individual differences in the acetabula of Asian patients with displaced femoral neck fractures. PATIENTS AND METHODS: Fifty patients with displaced femoral neck fractures were assessed (50 fractured hips, 50 non-fractured hips). On CT corrected by the anterior pelvic plane, the 100 hips were assessed regarding acetabular coverage (six parameters), acetabular depth (two parameters), and acetabular opening angle (four parameters). Additional parameters related to the fracture and sex were examined. The percentile of each parameter was shown for all hips. RESULTS: There was no patient with hip dysplasia defined as superior acetabular sector angle (SASA) less than 110° Compared with men, women had a significantly smaller anterior acetabular sector angle (AASA) (p = 0.016), and significantly larger acetabular inclination angle (p = 0.006) and acetabular index angle (p = 0.034). In the group with a normal SASA, seven hips (7.3%) had an anterior wall defect (AASA<50°) and five hips (5.2%) had a posterior wall defect (posterior acetabular sector angle<90°). CONCLUSION: Older adults with femoral neck fractures can have anterior wall and posterior wall defects, even if their SASA is normal. Hidden acetabular dysplasia may be related to post-BHA dislocation. So, our results suggest that is important to accurately evaluate the acetabulum of patients with femoral neck fracture before surgery.

Age-related visual outcomes in eyes with diffractive multifocal intraocular lenses.

BACKGROUND/OBJECTIVES: To assess the influence of patient age on visual outcomes in eyes with diffractive multifocal intraocular lenses (IOLs) SUBJECTS/METHODS: Based on age, we classified eyes with diffractive multifocal IOL into four groups: u50 (under 50 years old), 50s (50-59 years), 60s, and 70s. Corrected distance (CD), distance-corrected near (DCN) visual acuity (VA), and defocus curve were measured postoperatively. Using an "area-of-focus" metric, the distant, intermediate, and near area-of-focus (AoF) were also measured. These postoperative results were compared between the age groups. RESULTS: At 3 months after surgery, the CDVA in the u50, 50s, 60s, and 70s groups were -0.18, -0.16, -0.14, and -0.10 logMAR, respectively. The 70s CDVA was significantly worse than the u50 and 50s groups (P = 0.002, P = 0.049). The DCNVA in the u50, 50s, 60s, and 70s were 0.01, 0.03, 0.03, and 0.08 logMAR. DCNVA in the 70s group was significantly worse than that in the u50 and 60s groups (P = 0.008 and P = 0.019, respectively). The near AoF was smaller in the 70s than in the u50 and 50s groups (P = 0.040, P = 0.047). In both the intermediate and distant AoFs, there was no significant difference between the four age groups. A steep decline in near AoF was observed in patients over 60 years of age. CONCLUSIONS: The CDVA, DCNVA, and near AoF declined with patient age in eyes with diffractive multifocal IOL. The near AoF showed a drastic decline over 60 years.

Alcohol use patterns and risk of incident cataract surgery: a large scale case-control study in Japan.

To examine the risk of incident cataract surgery associated with alcohol use patterns among Japanese adults. This was a case-control study evaluating 14,861 patients with incident cataract surgery and 14,861 matched controls. Subjects admitted to any of the 34 hospitals in Japan and aged between 40 and 69 years were included. Drinking patterns (drinking frequency, daily average drinks, and total amount of lifetime drinking), smoking history, lifestyle-related comorbidities, and occupational factors were surveyed by trained interviewers. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using conditional logistic regression models. For drinking frequency, ORs in the 1-3 days/week and 4-7 days/week groups were 1.10 (95% CI 1.03-1.17) and 1.30 (1.21-1.40), respectively. For average drinks, ORs in > 0-2 drinks/day, > 2-4 drinks/day, and > 4 drinks/day were 1.13 (1.06-1.20), 1.23 (1.12-1.35), and 1.16 (1.03-1.31), respectively. Both men and women had an increased risk of incident cataract surgery with increased total lifetime drinking, with a significant increase in risk occurring at > 90 drink-years for men and > 40 drink-years for women. A positive dose-response relationship was observed between alcohol consumption and cataract. Restricted drinking may help to reduce the progression of cataracts.

Establishment of Rapid and Accurate Screening System for Molecular Target Therapy of Osteosarcoma.

Introduction Comprehensive analyses using clinical sequences subcategorized osteosarcoma (OS) into several groups according to the activated signaling pathways. Mutually exclusive co-occurrences of gene amplification (PDGFRA/KIT/KDR, VEGFA/CCND3, and MDM2/CDK4) have been identified in approximately 40% of OS, representing candidate subsets for clinical evaluation of additional therapeutic options. Thus, it would be desirable to evaluate the specific gene amplification before starting therapy in patients with OS. Materials and Methods This is a retrospective study. We examined 13 cases of clinical OS samples using NanoString-based copy number variation (CNV) analysis. Decalcification and chemotherapeutic effects on this analysis were also assessed. Results First, the accuracy of this system was validated by showing that amplification/deletion data obtained from this system using various types of cancer cell lines almost perfectly matched to that from the Cancer Cell Line Encyclopedia (CCLE). We identified potentially actionable alterations in CDK4/MDM2 amplification in 10% of samples and potential additional therapeutic targets (PDGFRA/KIT/KDR and VEGFA/CCND3) in 20% of samples, which is consistent with the reported frequencies. Furthermore, this assay could identify these potential therapeutic targets regardless of the sample status (frozen vs formalin-fixed paraffin-embedded [FFPE] tissues). Conclusion We established a NanoString-based rapid and cost-effective method with a short turnaround time (TAT) to examine gene amplification status in OS. This CNV analysis using FFPE samples is recommended where the histological evaluation of viable tumor cells is possible, especially for tumors after chemotherapy with higher chemotherapeutic effects.

A case of primary distal-type epithelioid sarcoma of the lumbar vertebra with a review of literature.

Epithelioid sarcoma (EpS) is a rare malignant neoplasm that accounts for < 1% of adult soft tissue sarcomas. Primary EpS of the bone is extremely rare and only a few cases have been reported to date. We report a case of primary distal-type EpS of the lumbar vertebra. A 30-year-old man without any history of malignant tumors had complained of lumbago for 3 months before visiting the hospital. Magnetic resonance imaging (MRI) of the lumbar spine showed a high signal intensity on the fat-suppressed T2-weighted image (WI) and a low signal on the T1WI at the L1 vertebral body. The tumor protruded toward the anterior components. Systemic radiological examination revealed no other lesion. A biopsy revealed a primary malignant tumor with epithelioid features. After chemotherapy, total en bloc spondylectomy was performed. Macroscopically, the tumor replaced the entire L1 with necrosis. Histologically, the tumor showed nodules of epithelioid cells that were strongly positive for epithelial markers, but a lack of INI1 expression. Central necrosis in the tumor nodule was also observed. This tumor showed loss of heterozygosity at the SMARCB1 locus but without the SMARCB1 mutation. The result of Foundation One ®CDx showed no actionable mutations. Seven months after surgery, a subcutaneous metastasis to the left cheek and bilateral lung metastasis with pleural dissemination were observed on radiological examination. A final diagnosis of distal-type EpS was made based on these findings. The patient died of the disease 8 months after surgery.

IRE1α-XBP1 but not PERK inhibition exerts anti-tumor activity in osteosarcoma.

Osteosarcoma (OS) is the most common primary malignant bone tumor. However, the therapeutic results of the advanced cases at the first visit were still extremely poor. Therefore, more effective therapeutic options based on molecular profiling of OS are needed. In this study, we investigated the functions of endoplasmic reticulum (ER) stress activities in OS and elucidated whether ER stress inhibitors could exert antitumor effects. The expression of 84 key genes associated with unfolded protein response (UPR) was assessed in four OS cells (143B, MG63, U2OS and KHOS) by RT2 Profiler PCR Arrays. Based on results, we performed both siRNA and inhibitor assays focusing on IRE1α-XBP1 and PERK pathways. All OS cell lines showed resistance to PERK inhibitors. Furthermore, ATF4 and EIF2A inhibition by siRNA did not affect the survival of OS cell lines. On the other hand, IRE1α-XBP1 inhibition by toyocamycin suppressed OS cell growth (IC50: < 0.075 µM) and cell viability was suppressed in all OS cell lines by silencing XBP1 expression. The expression of XBP1s and XBP1u in OS cell lines and OS surgical samples were confirmed using qPCR. In MG63 and U2OS, toyocamycin decreased the expression level of XBP1s induced by tunicamycin. On the other hand, in 143B and KHOS, stimulation by toyocamycin did not clearly change the expression level of XBP1s induced by tunicamycin. However, morphological apoptotic changes and caspase activation were observed in these two cell lines. Inhibition of the IRE1α-XBP1s pathway is expected to be a promising new target for OS.