Clinical characteristics of patients with not well-controlled severe asthma in Japan: Analysis of the Keio Severe Asthma Research Program in Japanese population (KEIO-SARP) registry.

BACKGROUND: Multiple phenotypes exist within the classification of severe asthma. However, characteristics of patients with not well-controlled severe asthma have not been well identified. METHODS: Japanese patients with asthma (age ≥ 20 years) were enrolled at the Keio University Hospital and its affiliated hospitals in this observational study (Keio Severe Asthma Research Program). Among them, patients with severe asthma (those undergoing Global Initiative for Asthma [GINA] 2018 step 4 or 5 treatment) were included in this analysis and investigated clinical characteristics based on asthma control status. RESULTS: Of the 154 patients (men, 46.8%; age, 60.1 ± 14.9 years), 87 (56.5%) had not well-controlled (partly controlled and uncontrolled) asthma (GINA step 4, 42 patients; step 5, 45 patients). Overall, there were no significant differences in clinical characteristics between patients with well-controlled and not well-controlled asthma. However, cluster analysis revealed that distinct 5 clusters (cluster 1, well-controlled; cluster 2, eosinophilic; cluster 3, non-type 2 inflammation; cluster 4, high periostin; and cluster 5, late-onset type 2 inflammation), and clusters 2-5 were not well-controlled. Among them, cluster 3 was characterized by low eosinophil counts, low periostin levels, and less frequent olfactory disturbance, and this cluster had the worst asthma control. CONCLUSIONS: Japanese patients with severe asthma were divided into well-controlled and not-well controlled asthma, and we confirmed heterogeneity of not well-controlled severe asthma. These patients, especially non-type 2 phenotype, require a further therapeutic approach. (University Hospital Medical Information Network Clinical Trials Registry, UMIN000002980).

Crystal structure of α-glucosyl transfer enzyme XgtA from Xanthomonas campestris WU-9701.

The α-glucosyl transfer enzyme XgtA is a novel type α-Glucosidase (EC 3.2.1.20) produced by Xanthomonas campestris WU-9701. One of the unique properties of XgtA is that it shows extremely high α-glucosylation activity toward alcoholic and phenolic -OH groups in compounds using maltose as an α-glucosyl donor and allows for the synthesis of various useful α-glucosides with high yields. XgtA shows no hydrolytic activity toward sucrose and no α-glucosylation activity toward saccharides to produce oligosaccharides. In this report, the crystal structure of XgtA was solved at 1.72 Å resolution. The crystal belonged to space group P22121, with unit-cell parameters a = 73.07, b = 83.48, and c = 180.79 Å. The ß→α loop 4 of XgtA, which is proximal to the catalytic center, formed a unique structure that is not observed in XgtA homologs. Furthermore, XgtA was found to contain unique amino acid residues around its catalytic center. The unique structure of XgtA provides an insight into the mechanism for the regulation of substrate specificity in this enzyme.

MicroRNA-27a regulates lipid metabolism and inhibits hepatitis C virus replication in human hepatoma cells.

The replication and infectivity of the lipotropic hepatitis C virus (HCV) are regulated by cellular lipid status. Among differentially expressed microRNAs (miRNAs), we found that miR-27a was preferentially expressed in HCV-infected liver over hepatitis B virus (HBV)-infected liver. Gene expression profiling of Huh-7.5 cells showed that miR-27a regulates lipid metabolism by targeting the lipid synthetic transcription factor RXRα and the lipid transporter ATP-binding cassette subfamily A member 1. In addition, miR-27a repressed the expression of many lipid metabolism-related genes, including FASN, SREBP1, SREBP2, PPARα, and PPARγ, as well as ApoA1, ApoB100, and ApoE3, which are essential for the production of infectious viral particles. miR-27a repression increased the cellular lipid content, decreased the buoyant density of HCV particles from 1.13 to 1.08 g/cm(3), and increased viral replication and infectivity. miR-27a overexpression substantially decreased viral infectivity. Furthermore, miR-27a enhanced in vitro interferon (IFN) signaling, and patients who expressed high levels of miR-27a in the liver showed a more favorable response to pegylated IFN and ribavirin combination therapy. Interestingly, the expression of miR-27a was upregulated by HCV infection and lipid overload through the adipocyte differentiation transcription factor C/EBPα. In turn, upregulated miR-27a repressed HCV infection and lipid storage in cells. Thus, this negative feedback mechanism might contribute to the maintenance of a low viral load and would be beneficial to the virus by allowing it to escape host immune surveillance and establish a persistent chronic HCV infection.

Effective elimination of contaminants after oral care in elderly institutionalized individuals.

After mechanical cleaning in oral care, eliminating residual oral contaminants has an important role in preventing their aspiration, especially in individuals with weak airway protection. We examined the effectiveness of wiping the oral cavity after oral care on eliminating contaminants in 31 patients who were hospitalized in our neurology inpatient unit. The amount of bacteria on the tongue, palate, and buccal vestibule was counted before and just after oral care, after eliminating contaminants either by rinsing with water and suction or by wiping with mouth wipes, and 1 h after oral care. Oral bacteria amounts were decreased significantly by both elimination procedures after oral care. These findings suggest that wiping with mouth wipes is as effective as mouth rinsing to decrease bacteria following oral care. With a lower risk of contaminant aspiration, wiping may be a suitable alternative to rinsing, especially in dysphagic individuals.

[A case of resection of pancreatic head lymph node metastasis of rectal carcinoid].

An 80-year-old woman was treated with low anterior resection and D3 lymph node dissection for rectal carcinoid in August 2007. Pathological diagnosis was a tumor 23 × 22 mm in size with muscularis propria invasion, ly1, v0, n1 (No. 251[1/12]). Partial hepatectomy was performed for liver metastasis in March 2010 and September 2011. In June 2013, an isolated tumor of 17 mm in diameter was detected between the inferior vena cava and the stomach using abdominal ultrasonography. The tumor was diagnosed as pancreatic head lymph node metastasis using abdominal enhanced computed tomography (CT), magnetic resonance cholangiopancreatography (MRCP), and positron emission tomography (PET); fluorodeoxyglucose (FDG) uptake was noted at the site of the tumor, but not at other parts of the body. Curative resection was the chosen treatment option. Open surgery was performed in September 2013, and a tumor 30 mm in diameter located at the ventral side of the inferior vena cava, lateral side of the portal vein, and cranial side of duodenum was resected. No surrounding tissue infiltration was found. Pathological analysis of the resected specimen indicated lymph node metastasis of the rectal carcinoid. The patient has been disease free for 9 months after surgery.

Unveiling the vitamin E profile in rice bran extracellular vesicles: evaluation of extraction and preparation methods.

Extracellular vesicles (EVs) are nanoscale entities secreted by various cells, encapsulating various nucleic acids and proteins that play important roles in cellular activities. Although rice bran is known for its richness in phytochemicals such as tocopherol and tocotrienol, the distribution of these compounds within EVs has not been extensively studied. The objective of this study was to detect and analyze the presence of vitamin E in EVs extracted from rice bran. We investigated several EV extraction methods, including rotation, vortex mixing, and ultrasonication, followed by post-extraction techniques such as ultracentrifugation, ultrafiltration, and lyophilization. Vitamin E in the EVs from rice bran was analyzed using LC-FLD. This study is the first to identify tocopherol and tocotrienol in rice bran-derived EVs. Our results indicate that ultracentrifugation followed by rotation is the most effective method for the preparation of rice bran-derived EVs. Notably, the vitamin E profile in EVs varies depending on the preparation method and differs from that in rice bran extracts. The pronounced presence of vitamin E in EVs suggests unique pharmacokinetics and underscores the potential of EVs as carriers for drug delivery systems. This study not only confirms the presence of vitamin E in EVs, but also underscores the potential of EVs and their phytochemical content for therapeutic applications.

[Long-term survival of a patient with locally advanced unresectable pancreatic cancer treated with gemcitabine after chemoradiation therapy].

We report a case of long-term survival of a patient with locally advanced unresectable pancreatic cancer treated with gemcitabine after chemoradiation therapy( CRTx). A 61-year-old woman was diagnosed as having locally advanced unresectable pancreatic cancer, 3.0 cm in diameter, which had invaded the superior mesenteric artery by computed tomography (CT). She was treated with CRTx( gemcitabine[ GEM] at 250 mg/m2/week for 6 weeks+liniac irradiation of 50.2 Gy) followed by systemic chemotherapy (gemcitabine (GEM) at 1,000 mg/m2). Reassessment after CRTx yielded a diagnosis of stable disease (according to the Response Evaluation Criteria in Solid Tumors [RECIST] 1.1). To date, the patient has undergone 57 courses of chemotherapy in the outpatient clinic; however, she did require biliary stent placement because of occlusive jaundice.

[A case of SPT therapy( S-1+CDDP+trastuzumab) was successful for HER2-positive gastric cancer accompanied by distant lymph node metastasis].

A 63-year-old man visited our clinic with a chief complaint of a left axillary mass, and after a series of examinations, gastric cancer was diagnosed. Histopathological examination revealed a human epidermal growth factor receptor (HER)-2- positive( immunohistochemistry[ IHC], 2+; fluorescence in situ hybridization[ FISH], positive) poorly differentiated adenocarcinoma, and SPT therapy( S-1 at 120 mg/m2/day+cisplatin[ CDDP] at 60 mg/m2+trastuzumab at 8 mg/kg) was initiated. The primary lesion and swollen lymph nodes initially decreased markedly in size; however, the lymph node swelling worsened at 186 days( 6.2 months) after the initiation of treatment. Progressive disease( PD) was diagnosed, and SPT therapy was terminated. One cycle of S-1+docetaxel was administered as second-line therapy, but the regimen was changed to docetaxel monotherapy due to adverse effects. After 5 cycles of this treatment, the primary lesion had decreased in size and the lymphadenopathy disappeared. Positron emission tomography( PET)-computed tomography (CT) revealed fluorodeoxyglucose( F18)( FDG) accumulation only in the primary lesion, and therefore, with the patient's informed consent distal gastrectomy was performed approximately 14 months after the first treatment. The postoperative diagnosis was gastric cancer fStage IA (T1a, N0, and M0). Here, we discuss a case of HER2-positive gastric cancer with references. Among the advanced recurrent gastric cancers tested between April 2011 and February 2013, 16.4% (11/67) were HER2-positive.

[Long-term survival after surgical resection of a cancer of unknown primary site-a case report].

We report a case of long-term survival after combination chemotherapy and surgical resection of a cancer of unknown primary site[ CUPs]. A septuagenarian female was identified as having high blood levels of carcinoembryonic antigen (CEA) during follow-up monitoring of asthma. Endoscopy and imaging studies including computed tomography (CT) and positron emission tomography (PET)-CT revealed a malignant lymph node adjacent to the abdominal aorta; however, no other lesion was detected. Therefore, we performed CT-guided biopsy and diagnosed the lesion to be a lymph node metastasis of poorly differentiated adenocarcinoma. As we considered this as a systemic disease, the patient received 2 courses of combination chemotherapy with 5-fluorouracil( 5-FU)/cisplatin( CDDP) and achieved a partial response (PR). Later, the patient received S-1 therapy as second-line chemotherapy and S-1/irinotecan( CPT-11) as third-line chemotherapy in an outpatient clinic. However, the tumor continued to grow, and therefore, we decided to perform surgical resection. Histopathological examination of the resected specimen yielded a diagnosis of metastatic adenocarcinoma of the lymph node. The patient has been well without any signs of recurrence for more than 9 years since surgery. As CUPs is generally associated with poor prognosis, this case raises the possibility that combination therapy might improve convalescence.

Enzymatic and selective production of alkyl α-d-glucopyranosides by the α-glucosyl transfer enzyme derived from Xanthomonas campestris WU-9701.

Several alkyl glucosides exhibit various bioactivities. 1-Octyl ß-d-glucopyranoside produced by organic synthesis is used as a nonionic surfactant. However, no convenient method has been developed for the selective production of alkyl α-glucosides (α-AGs), such as 1-octyl α-d-glucopyranoside (α-OG). Therefore, we developed a simple method for selective production of α-AGs using the glucosyl transfer enzyme XgtA, (E.C. 3.2.1.20), derived from Xanthomonas campestris WU-9701. When 0.80 M alkyl alcohol and 2.5 units XgtA were incubated in 2.0 mL of 30 mM HEPES-NaOH buffer (pH 8.0) containing 1.2 M maltose at 45 °C, a specific α-AG corresponding to each alkyl alcohol (C2-C10) was detected. Under the standard conditions, we examined the selective production of α-OG from 1-octanol and maltose using XgtA. The reaction product was isolated and identified as α-OG via 1H nuclear magnetic resonance and nuclear overhauser effect spectroscopy analyses. No other glucosylated products, such as maltotriose, were detected in the reaction mixture. Under the standard conditions at 45 °C for 96 h, 243 mM α-OG (71 g/L) was produced in one batch production. Moreover, the addition of glucose isomerase to the reaction mixture decreased the concentration of glucose released via the reaction and increased the amount of α-OG produced; 359 mM α-OG (105 g/L) was maximally produced at 96 h. In conclusion, this study demonstrates the selective production of α-AGs using a simple enzymatic reaction, and XgtA has the potential to selectively produce various α-AGs.

Identification of epigenetically active L1 promoters in the human brain and their relationship with psychiatric disorders.

Long interspersed nuclear element-1 (LINE-1, L1) affects the transcriptome landscape in multiple ways. Promoter activity within its 5'UTR plays a critical role in regulating diverse L1 activities. However, the epigenetic status of L1 promoters in adult brain cells and their relationship with psychiatric disorders remain poorly understood. Here, we examined DNA methylation and hydroxymethylation of the full-length L1s in neurons and nonneurons and identified "epigenetically active" L1s. Notably, some of epigenetically active L1s were retrotransposition competent, which even had chimeric transcripts from the antisense promoters at their 5'UTRs. We also identified differentially methylated L1s in the prefrontal cortices of patients with psychiatric disorders. In nonneurons of bipolar disorder patients, one L1 was significantly hypomethylated and showed an inverse correlation with the expression level of the overlapping gene NREP. Finally, we observed that altered DNA methylation levels of L1 in patients with psychiatric disorders were not affected by the surrounding genomic regions but originated from the L1 sequences. These results suggested that altered epigenetic regulation of the L1 5'UTR in the brain was involved in the pathophysiology of psychiatric disorders.

Diagnostic and prognostic biomarkers for progressive fibrosing interstitial lung disease.

No biomarkers have been identified in bronchoalveolar lavage fluid (BALF) for predicting fibrosis progression or prognosis in progressive fibrosing interstitial lung disease (PF-ILD). We investigated BALF biomarkers for PF-ILD diagnosis and prognosis assessment. Overall, 120 patients with interstitial pneumonia who could be diagnosed with PF-ILD or non PF-ILD were enrolled in this retrospective study. PF-ILD was diagnosed according to Cottin's definition. All patients underwent bronchoscopy and BALF collection. We evaluated blood and BALF parameters, high-resolution computed tomography (HRCT) patterns, and spirometry data to identify factors influencing PF-ILD diagnosis and prognosis. On univariate logistic analysis, age, sex, the BALF white blood cell fraction (neutrophil, lymphocyte, eosinophil, and neutrophil-to-lymphocyte ratio), BALF flow cytometric analysis (CD8), and an idiopathic pulmonary fibrosis/usual interstitial pneumonia pattern on HRCT were correlated with PF-ILD diagnosis. Multivariate logistic regression analysis revealed that sex (male), age (cut-off 62 years, area under the curve [AUC] 0.67; sensitivity 0.80; specificity 0.47), white blood cell fraction in BALF (NLR, neutrophil, and lymphocyte), and CD8 in BALF (cut-off 34.2; AUC 0.66; sensitivity, 0.74; specificity, 0.62) were independent diagnostic predictors for PF-ILD. In BALF, the NLR (cut-off 8.70, AUC 0.62; sensitivity 0.62; specificity 0.70), neutrophil count (cut-off 3.0, AUC 0.59; sensitivity 0.57; specificity 0.63), and lymphocyte count (cut-off 42.0, AUC 0.63; sensitivity 0.77; specificity 0.53) were independent diagnostic predictors. In PF-ILD patients (n = 77), lactate dehydrogenase (cut-off 275, AUC 0.69; sensitivity 0.57; specificity 0.78), Krebs von den Lungen-6 (cut-off 1,140, AUC 0.74; sensitivity 0.71; specificity 0.76), baseline forced vital capacity (FVC) (cut-off 1.75 L, AUC 0.71; sensitivity, 0.93; specificity, 0.46), and BALF neutrophil ratio (cut-off 6.0, AUC 0.72; sensitivity 0.79; specificity 0.80) correlated with death within 3 years. The BALF cellular ratio, particularly the neutrophil ratio, correlated with the diagnosis and prognosis of PF-ILD. These findings may be useful in the management of patients with interstitial pneumonia.

[Utility of the WallFlexTM duodenal stent for unresectable advanced gastric cancer related to gastric outlet obstruction].

Duodenal stenting for malignant disease related to gastric outlet obstruction(GOO) has been covered by health insurance in Japan since April 2010. We inserted WallFlexTM duodenal stents(WDS) in 4 patients with GOO caused by unresectable gastric cancer. WDS insertion was successful in all 4 cases. Duodenal perforation occurred in 1 case. One case each of stent obstruction and stent migration occurred. All patients could eat a soft-food diet for 3-6 months (median, 5.3 months). Survival time ranged between 5 and 14 months (median, 6 months). Three patients underwent S-1 combination chemotherapy. Duodenal stenting is expected to be effective for advanced gastric cancer related to GOO.

[Two case reports of neuroendocrine cell carcinoma of the stomach].

We encountered 2 cases of endocrine cell carcinoma of the stomach. One patient had a type 3 tumor in the cardia, and the other patient had a submucosal tumor in the posterior wall of the gastric angle, both of which were detected by upper gastrointestinal endoscopy. The first patient underwent total gastrectomy(D1+No.7, 19) with cholecystectomy. Microscopic histological examination fortuitously revealed a tumor thrombus in the vessel of the gallbladder. Eight months after the operation, a paraaortic lymph node recurrence was detected, and the patient died 28 months after the operation. The second patient underwent distal gastrectomy, cholecystectomy, S5 subsegmental hepatectomy, and S8 radiofrequency ablation(RFA). Despite undergoing adjuvant chemotherapy[irinotecan(CPT-11)/cisplatin(CDDP)], multiple liver recurrences were detected 6 months after the operation. Consequently, systemic chemotherapy (S-1/CDDP) was performed.

Upregulation of IL-4 receptor signaling pathway in circulating ILC2s from asthma patients.

Background: Group 2 innate lymphoid cells (ILC2s) produce type 2 cytokines by stimulation with epithelial cell-derived cytokines and are implicated in the pathogenesis of various allergic diseases, including asthma. However, differences in the molecular characteristics of ILC2s between patients with asthma and healthy subjects remain unclear. Objective: We sought to evaluate differences in cytokine production capacity and gene expression profile of ILC2s in the peripheral blood of patients with asthma and healthy subjects. Methods: We evaluated ILC2s derived from 15 patients with asthma and 7 healthy subjects using flow cytometry, live-cell imaging of secretion activity analysis, and RNA-sequencing. Results: ILC2s were sorted as CD45+Lineage-CRTH2+CD127+CD161+ cells from the peripheral blood of patients with asthma and healthy subjects, and the number of ILC2s was decreased in patients with asthma (851 ± 1134 vs 2679 ± 3009 cells/20 mL blood; P = .0066). However, patient-derived ILC2s were activated to produce more IL-5 and IL-13 in response to stimulation with IL-2, IL-33, and thymic stromal lymphopoietin compared with healthy subject-derived ILC2s (P = .0032 and P = .0085, respectively). Furthermore, RNA-sequencing analysis revealed that patient-derived ILC2s had different gene expression profiles, such as increased expression in cell growth-related genes (CDKN1b, CCNG2, CCND2, CCN1), prostaglandin E receptor (PTGER2), and IL-4 receptor. In addition, a gene set of the IL-4 receptor signaling pathway was significantly upregulated in ILC2s in patients with asthma (P = .042). Conclusions: Our results suggest that circulating ILC2s in patients with asthma are preactivated via the IL-4 receptor signaling pathway and produce IL-5 and IL-13 vigorously by stimulation.

[A case of S-1/CDDP chemotherapy for inoperable advanced gastric cancer which led to gastrectomy with histological complete response].

As the treatment for inoperable advanced gastric cancer, S-1/CDDP combination therapy (SP chemotherapy) has become a standard treatment. In our hospital, a second course of chemotherapy was performed on an outpatient basis in order to improve a traditional QOL. In this case, it showed remarkable effects in 15 months after starting chemotherapy. Then gastrectomy was performed. Histological findings of the resected specimens confirmed pCR in all tumors. We report on progress of this case and explain about the ingenuity of SP chemotherapy.