Intracellular metabolite ß-glucosylceramide is an endogenous Mincle ligand possessing immunostimulatory activity.

Sensing and reacting to tissue damage is a fundamental function of immune systems. Macrophage inducible C-type lectin (Mincle) is an activating C-type lectin receptor that senses damaged cells. Notably, Mincle also recognizes glycolipid ligands on pathogens. To elucidate endogenous glycolipids ligands derived from damaged cells, we fractionated supernatants from damaged cells and identified a lipophilic component that activates reporter cells expressing Mincle. Mass spectrometry and NMR spectroscopy identified the component structure as ß-glucosylceramide (GlcCer), which is a ubiquitous intracellular metabolite. Synthetic ß-GlcCer activated myeloid cells and induced production of inflammatory cytokines; this production was abrogated in Mincle-deficient cells. Sterile inflammation induced by excessive cell death in the thymus was exacerbated by hematopoietic-specific deletion of degrading enzyme of ß-GlcCer (ß-glucosylceramidase, GBA1). However, this enhanced inflammation was ameliorated in a Mincle-deficient background. GBA1-deficient dendritic cells (DCs) in which ß-GlcCer accumulates triggered antigen-specific T-cell responses more efficiently than WT DCs, whereas these responses were compromised in DCs from GBA1 × Mincle double-deficient mice. These results suggest that ß-GlcCer is an endogenous ligand for Mincle and possesses immunostimulatory activity.

[A Case of Consciousness Disorder Induced by the Syndrome of Inappropriate Antidiuretic Hormone Secretion Following Cisplatin and 5-Fluorouracil Chemotherapy in a Patient with Tongue Cancer].

We herein report a case of a consciousness disorder that was induced by the syndrome of inappropriate antidiuretic hormone secretion following cisplatin (CDDP) and 5 -fluorouracil (5-FU) chemotherapy in a patient with tongue cancer. A 72- year-old woman complained of tongue pain and was admitted to our hospital for neoadjuvant chemotherapy, under a diagnosis of tongue squamous cell carcinoma (T4aN2bM0). She was treated with CDDP and 5-FU. On the second day after administration, she complained of nausea and anorexia, and on the third day, she showed impaired consciousness. Laboratory studies revealed that the patient had a serum sodium concentration 112mEq/L, and no dehydration was noted. The patient was diagnosed with SIADH, using the appropriate diagnostic criteria based on serum and urine hypoosmolality. We subsequently discontinued chemotherapy and initiated fluid restriction and sodium supplements. Two days after this treatment, her consciousness level improved, and on the fifth day of treatment, laboratory studies revealed a serum sodium level of 134mEq/ L.

Clinical Characteristics of the Cleft Lip and/or Palate: Association with Congenital Anomalies, Syndromes, and Chromosomal Anomalies.

Background: Cleft lip and/or palate (CL/P) can be accompanied by other congenital anomalies. We conducted a long-term evaluation of the associations between cleft patterns, sex distribution, and accompanying congenital anomalies of patients with CL/P. Methods: The medical records of 739 patients with CL/P, seen between January 1967 and December 2020, were retrospectively reviewed. Fisher's exact test was used for statistical analysis. Results: Among the 739 patients with CL/P, the male-to-female ratio was 1.1. Regarding the cleft pattern, 121 (16.4%), 104 (14.1%), 280 (37.9%), 198 (26.8%), and 36 (4.9%) patients had cleft lip (CL), cleft lip and alveolus (CLA), cleft lip and palate (CLP), cleft palate (CP), and submucous cleft palate (SMCP), respectively. Congenital anomalies were identified in 107 (14.5%) cases, of which 53 (49.5%) had congenital heart disease. The frequencies of congenital anomalies patients with in CL/P were 14/225 (6.2%), 36/280 (12.9%), 43/198 (21.7%), and 14/36 (38.9%) for a combination of CL and CLA, CLP, CP, and SMCP, respectively. Accompanying syndromes and chromosomal anomalies were identified in 40 (5.4%) cases, in which Pierre Robin sequence (16 cases of CP and 4 cases of SMCP) was the most frequent. Conclusion: No sex differences were observed in CL/P, and CLP and CP were the most common cleft patterns. Congenital anomalies associated with CL/P were dominated by congenital heart disease and were most frequently identified in CP and SMCP cases. Notably, the Pierre Robin sequence, a complex syndrome characterized by micrognathia, glossoptosis, respiratory obstruction, and a U- or V-shaped CP, was found in cases of both CP and SMCP, and accounted for the symptoms in most cases.

Isolation of tumor-infiltrating lymphocytes from preserved human tumor tissue specimens for downstream characterization.

Immunophenotyping of tumor-infiltrating lymphocytes (TILs) by flow cytometry can predict clinical efficacy of immunotherapy. However, several obstacles need to be overcome for developing a flow cytometry assay starting from solid tumor specimens. Here, we show a detailed enzyme-based protocol to isolate TILs from human tumor tissues. The protocol was optimized to obtain enough viable TILs from a biopsy tissue specimen for flow cytometry-based TIL immunophenotyping. Additionally, tissue samples could be preserved for up to 72 h for subsequent characterization. For complete details on the use and execution of this protocol, please refer to Kumagai et al. (2020, 2022).

Clinical Features of Oral Multiple Primary Carcinomas Compared with Oral Single Primary Carcinoma.

BACKGROUND: Owing to the increase in the older population and the increased life span, the number of patients with oral multiple primary carcinomas will increase. Predicting the second and third carcinoma clinically is difficult, and the presence of second or third carcinomas is a factor that determines the prognosis of oral carcinoma. In this study, we examined the clinical features of oral multiple primary carcinomas treated in our department. METHODS: We retrospectively reviewed the medical records of patients with oral squamous cell carcinoma who underwent radical treatment at and were followed by the Department of Oral and Maxillofacial Surgery, Tottori University Hospital from January 2003 to October 2017. RESULTS: This study included 261 patients: 241 patients had oral single primary carcinoma and 20 patients had oral multiple primary carcinomas. Oral multiple primary carcinomas showed female predilection and occurred more frequently in the lower gingiva and significantly less frequently in the tongue (P < 0.01). Oral multiple primary carcinomas showed a significantly higher recurrence rate (P < 0.01). The 5-year overall survival of oral single primary carcinoma patients was 88.0% compared with 95% for oral multiple primary carcinomas, with no significant difference (log rank test, P = 0.54). However, the 15-year survival rate dropped to 28.1% in oral multiple primary carcinomas. The cumulative disease incidence rates of metachronous second primary carcinoma from first carcinoma at 5 years and 10 years were 3.45% and 5.36%, respectively. CONCLUSION: Oral multiple primary carcinomas rarely occur in the tongue. The 5-year survival rate showed no difference between single and multiple carcinoma patients, but over longer observation, the prognosis of multiple carcinoma was poor owing to a high recurrence rate. Because of the high recurrence rate and risk of further metachronous carcinoma in oral multiple primary carcinomas, longer-term follow-up is required.

Study on the Development of a New Device with Dual Cameras for Evaluating Expiratory Nasal Flow.

BACKGROUND: Use of the Glatzel mirror for measuring expiratory nasal flow in preschool children has the disadvantage of vagueness, and the mirror may induce fear and inhibition of interest in those children. In response to these limitations, we developed a new device with dual cameras for measuring expiratory nasal flow in 2 to 6 year old children. The aim of this study is to compare the Glatzel mirror and the new device, in terms of accurate assessment of expiratory nasal flow, children's feelings, and correlation to each child's profile. METHODS: This study evaluated 20 cleft lip and palate patients and 21 healthy children aged between 2 and 6 (under 7) years. After consent was granted, a 4-week screening period was undertaken followed by inspection at weeks 8, 16, 24, and 32. Each inspection was conducted while the children were asked to pronounce various sounds and comprised three stages: i) use of the Glatzel mirror, ii) subjective visual assessment using the new device, and iii) image recording by dual cameras of the new device. Questionnaires for the new device were administered at the initial and final inspections. To contrast the results between the Glatzel mirror and the new device, the numbers that indicated values of subjective visual assessment and camera assessment greater than the assessment values of the Glatzel mirror were compared. For measuring the children's responses to the new device compared with those to the Glatzel mirror, the answers to the questionnaires were compared. For the comparison of the children's profiles (age and sex) and feelings, the numbers of subjects who could use the new device were measured. RESULTS: The camera assessment of the new device indicated significantly greater values than that of the Glatzel mirror (P < 0.05). The feelings of the subjects to the new device mostly improved as the study progressed. Subjects aged 3 years and older were generally able to use the new device from the initial inspection. For both sexes, as the inspection progressed, the number occasions of successful use increased. CONCLUSION: This study demonstrated the superiority of the new device with dual cameras to the Glatzel mirror in terms of functionality and attitude of children.

A novel PAX3 mutation in a Japanese boy with Waardenburg syndrome type 1.

Waardenburg syndrome type 1 (WS1) is a rare autosomal dominant disorder characterized by hair hypopigmentation, abnormal iris pigmentation, and congenital hearing loss. WS1 is caused by mutations in paired box gene 3 (PAX3). We identified a novel PAX3 mutation (c.1107 C>G, p.Ser369Arg) in a Japanese WS1 patient showing abnormal right iris pigmentation, right-sided congenital hearing loss, synophrys, incomplete left cleft lip, and cryptorchidism.