Search

A 75-year-old man with a fever, shoulder pain, and lower leg edema was diagnosed with polymyalgia rheumatica and started on glucocorticoid therapy. Eighteen months later, he was admitted with rapidly progressive renal failure. Glucocorticoid therapy had been discontinued one month prior to admission. Serum anti-glomerular basement membrane antibodies were elevated, and a kidney biopsy revealed fibrinoid necrosis of the medium-sized renal arteries, tubulointerstitial nephritis, and collapsed glomeruli. An immunofluorescence study showed mild immunoglobulin G linear deposition. Polyarteritis nodosa was diagnosed based on the presence of fibrinoid necrosis in the medium-sized renal arteries. Glucocorticoid pulse therapy and plasmapheresis were initiated, but the patient died of alveolar hemorrhaging. This was a rare case of Goodpasture syndrome with polyarteritis nodosa.

Extracellular vesicles (EVs) are generated in all cells. Systemic administration of allogenic EVs derived from epithelial and mesenchymal cells has been shown to be safe, despite carrying an array of functional molecules, including thousands of proteins. To address whether epithelial cell-derived EVs can be modified to acquire the capacity to induce an immune response, we engineered 293T EVs to harbor the immunomodulatory molecules CD80, OX40L, and PD-L1. We demonstrated abundant levels of these proteins in the engineered cells and EVs. Functionally, the engineered EVs efficiently elicited positive and negative costimulation of human and murine T cells. In the setting of cancer and autoimmune hepatitis, the engineered EVs modulated T cell functions and altered disease progression. OX40L EVs also provided enhanced antitumor activity in combination with anti-CTLA-4 in melanoma-bearing mice. In addition, we added multiple immunomodulatory proteins in EVs (EV mIM ), attempting to elicit an immune response in both lymphoid and myeloid compartments. The EV mIM containing CD80, 4-1BBL, CD40L, CD2, and CD32 engaged both T cells and antigen presenting cells (APCs) in melanoma tumors, demonstrating the capacity for EV mIM to elicit antitumor activity. Our work provides evidence that EVs can be engineered to induce specific immune responses with translational potential to modulate immune cell functions in pathological settings.

The 18 th Workshop on Recent Issues in Bioanalysis (18 th WRIB) took place in San Antonio, TX, USA on May 6-10, 2024. Over 1100 professionals representing pharma/biotech companies, CROs, and multiple regulatory agencies convened to actively discuss the most current topics of interest in bioanalysis. The 18 th WRIB included 3 Main Workshops and 7 Specialized Workshops that together spanned 1 week to allow an exhaustive and thorough coverage of all major issues in bioanalysis of biomarkers, immunogenicity, gene therapy, cell therapy and vaccines.Moreover, in-depth workshops on "IVDR Implementation in EU & Changes for LDT in the US" and on "Harmonization of Vaccine Clinical Assays Validation" were the special features of the 18 th edition.As in previous years, WRIB continued to gather a wide diversity of international, industry opinion leaders and regulatory authority experts working on both small and large molecules as well as gene, cell therapies and vaccines to facilitate sharing and discussions focused on improving quality, increasing regulatory compliance, and achieving scientific excellence on bioanalytical issues.This 2024 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2024 edition of this comprehensive White Paper has been divided into three parts for editorial reasons.This publication (Part 1) covers in Part 1A the Recommendations on Mass Spectrometry Assays and Regulated Bioanalysis/BMV and in Part 1B the Regulatory Inputs on these topics. Part 3 (Gene Therapy, Cell therapy, Vaccines and Biotherapeutics Immunogenicity) and Part 2 (Biomarkers/BAV, IVD/CDx, LBA and Cell-Based Assays) are published in volume 17 of Bioanalysis, issues 3 and 4 (2025), respectively.

Objectives: To evaluate the efficacy and safety of first-line targeted synthetic disease-modifying anti-rheumatic drugs (tsDMARDs) in patients with rheumatoid arthritis (RA) and chronic kidney disease (CKD)., Methods: This retrospective cohort study included 216 patients with RA prescribed their first tsDMARDs at two hospitals between 2013 and 2022. Dose reduction and contraindication guidelines for tsDMARDs according to kidney function were followed. The patients were categorised by kidney function and tsDMARD modality. The primary outcome was the 24-month drug retention rate, and the secondary outcomes were changes in the Disease Activity Score 28-C-reactive protein (DAS28-CRP) level, prednisolone dosage, and reasons for discontinuation., Results: The 24-month drug retention rates according to the estimated glomerular filtration rate (eGFR) (≥60, 30-60, or < 30 mL/min/1.73 m2) were as follows: all tsDMARDs (46.0, 44.1, 47.1%), tofacitinib (55.9, 53.3, 66.7%), baricitinib (64.2, 42.0%), and peficitinib (36.4, 44.1, 40.0%). Even in groups with lower kidney function, the drug retention rate was maintained (adjusted hazard ratio was 1.14 (95% confidence interval, 0.81-1.62), p= 0.45). Patients had a decreased DAS28-CRP (p< 0.01) and a reduced prednisolone dosage (p< 0.01) over the six-month period following tsDMARD initiation. The incidence of herpes zoster and deep vein thrombosis (DVT) was higher in the group with an eGFR <30 mL/min/1.73 m2, but not statistically significant., Conclusions: tsDMARDs have demonstrated efficacy and safety in patients with RA and CKD; however, clinicians should consider the potential for herpes zoster and DVT in patients with eGFR <30 mL/min/1.73 m2., (© The Author(s) 2025. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: The clinical outcomes of percutaneous coronary intervention (PCI) using drug-coated balloons (DCB) for de novo coronary artery lesions with eruptive calcified nodules remain unclear., Methods and Results: This retrospective study analyzed the long-term outcomes of 308 consecutive patients (389 lesions) treated with PCI using DCB under optical coherence tomography guidance for de novo coronary artery lesions between September 2018 and November 2020. Patients were classified into 2 groups: those with an eruptive calcified nodule in the culprit lesion (CN group) and those without (non-CN group). The primary endpoint was major adverse cardiovascular events (MACE), including clinically driven target lesion revascularization (TLR), myocardial infarction (MI), and cardiac death. The median follow-up period was 2.6 years (interquartile range 1.9-3.4 years). The CN group had significantly higher rates of MACE (hazard ratio [HR] 9.2; 95% confidence interval [CI] 4.1-20.2; P<0.0001), TLR (HR 5.0; 95% CI 1.7-15.1; P<0.01), MI (HR 30.5; 95% CI 5.0-184.8; P<0.001), and cardiac death (HR 25.1; 95% CI 8.7-72.6; P<0.0001) than the non-CN group. Results were similar even after adjusting for potential confounding factors using propensity score matching., Conclusions: This study demonstrated that patients with eruptive calcified nodules who underwent PCI with DCB for de novo coronary artery lesions had worse long-term clinical outcomes than patients without such nodules.

Background: Cancer immune responses are generated in secondary lymphoid organs, such as the lymph nodes and tonsils. In the current study, transcriptional profiles of peritumoral tonsillar tissues (PTTs) from oropharyngeal cancers (OPCs) were assessed and compared with those of inflammatory tonsils and regional lymph nodes (rLNs)., Methods: RNA samples of PTTs and rLNs from 13 OPCs, and 4 inflammatory tonsils were subjected to microarray analysis, and differentially expressed genes (DEGs) identified from 730 nCounter Panel immune-related genes. Gene Set enrichment Analysis (GSEA) was used for DEG profiling of PTTs and rLNs between lymph node metastasis-negative and metastasis-positive cases. The top 20 genes, as ranked by GSEA metric scores, were extracted and subjected to principal component analysis (PCA). The correlation of each patient's PCA score with lymph node status was assessed by Receiver Operating Characteristics (ROC) analysis., Results: Comparing DEG analyses of PTTs with those of inflammatory tonsils and rLNs revealed 144 and 45 upregulated genes, respectively. ClueGO, a widely used Cytoscape plug-in, revealed activated pathways in PTTs, including lymphocyte proliferation (followed by T cell activation involved in the immune response) and positive regulation of leukocyte migration (followed by antimicrobial humoral immune response mediated by antimicrobial peptides) as the most significantly enriched immune system process functions in the gene ontology when comparing inflammatory tonsils and rLNs. The area under the ROC curves of PTTs and rLNs were 0.806 and 0.389, and were significant by DeLong's test (p = 0.025)., Conclusion: PTTs exhibit unique immunological features distinguishing them from inflammatory tonsils and rLNs. Gene expression analysis of PTTs is useful for investigating the mechanism of OPC lymphatic spread, even compared with analysis of rLNs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2025 Wakisaka et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Phospholipids are the most abundant component in lipid membranes and are essential for the structural and functional integrity of the cell. In eukaryotic cells, phospholipids are primarily synthesized de novo through the Kennedy pathway that involves multiple enzymatic processes. The terminal reaction is mediated by a group of cytidine-5'-diphosphate (CDP)-choline /CDP-ethanolamine-phosphotransferases (CPT/EPT) that use 1,2-diacylglycerol (DAG) and CDP-choline or CDP-ethanolamine to produce phosphatidylcholine (PC) or phosphatidylethanolamine (PE) that are the main phospholipids in eukaryotic cells. Here we present the structure of the yeast CPT1 in multiple substrate-bound states. Structural and functional analysis of these binding-sites reveal the critical residues for the DAG acyl-chain preference and the choline/ethanolamine selectivity. Additionally, we present the structure in complex with a potent inhibitor characterized in this study. The ensemble of structures allows us to propose the reaction mechanism for phospholipid biosynthesis by the family of CDP-alcohol phosphotransferases (CDP-APs)., Competing Interests: Competing interests: S.M. is an employee of Xaira Therapeutics. The remaining authors declare no competing interests., (© 2024. The Author(s).)

Emphysematous polycystic renal infection (EPRI) has a poor prognosis with conservative management, and early surgical nephrectomy has been recommended. However, percutaneous cyst drainage may be a possible treatment option. We experienced 6 patients with autosomal dominant polycystic kidney disease (ADPKD) presenting with EPRI. Three patients developed EPRI after renal transarterial embolization (TAE), and the other 3 developed EPRI independently of renal TAE. Two of the patients had only one cyst with gas formation, and the causative organism was sensitive to antibiotics; these patients were cured by cyst drainage and antibiotic therapy. However, in 3 patients with severe renal enlargement and gas formation in multiple cysts, the causative organism was antibiotic resistant and cyst drainage was not effective, so surgical nephrectomy was performed and the disease effectively treated. Surgical nephrectomy should be considered in patients with multiple cysts with gas formation and severe renal enlargement., Competing Interests: The authors have no competing financial interests or conflicts of interest to declare. Figure 1Computed tomography images. a: Case 1: Horizontal surface border structure between fluid and gas (arrow) was seen only in one 10-cm cyst in the upper pole of the right kidney. b: Case 2: Significant gas formation (arrow) was found in only one 5 × 6 cm large cyst in the right kidney. c: Case 3: More than three cysts with gas formation were seen in the right kidney. d: Case 4: Gas formation as observed in multiple cysts of the right kidney. e: Case 5: Gas formation was seen in numerous cysts of the right kidney. f: Case 6: Gas formation was seen in numerous cysts of both kidneys. Table 1.The table summarizes clinical information for six cases. Case123456Age (years)775664527468SexWomanWomanManManManWomanYears on dialysisCKD stage4 (eGFR 26.3)CKD stage 4 (eGFR 11.2)CAPD 3 yearsCAPD4 years + HD 3 yearsHD 1 yearHD 6 yearsTriggersPreceding cystitisUnknownUnknownAfter renal TAEAfter renal TAEAfter renal TAEAir formation (right kidney)One cystOne cystAt least 3 cystsMany cystMany cystMany cystsAir formation (left kidney)NoneNoneNoneNoneSmall cystMany cystsCausative agentMultidrug-sensitive Klebsiella pneumoniaeMultidrug-sensitive E. coliMultidrug-sensitive Klebsiella pneumoniaeMultidrug-resistant E. coliNot inspectedMultidrug-resistant E. coli and Candida AlbicaneCyst drainageYes (three times)Yes (1 time)Yes (1 time)Yes (1 time)NoYes (3 times)Surgical nephrectomyNoneNoneRight nephrectomyRight nephrectomyBoth kidneysNoPrognosisSurviveSurviveSurviveSurviveSurviveDeathHistory of diabetesYesYesNoNoNoNoHistory of urinary tract infectionYesYesNoNoNoNo Figure 2This describes the treatment course, renal function, and CRP trends in Case 1. The patient’s overall condition improved after multiple drainages.  , (© Dustri-Verlag Dr. K. Feistle.)

A 47-year-old woman with a 12-year history of anemia and high C-reactive protein (CRP) levels was admitted to our hospital with worsening fatigue and night sweats. She had high levels of immunoglobulin G (IgG; 4182 mg/dL), IgA (630.6 mg/dL), and CRP (7.44 mg/dL); a low hemoglobin level (8.9 g/dL); urinary protein (11.83 g/day); and urinary sediment (20 - 29 red blood cells per high power field). On the basis of the clinical findings and biopsied lymph nodes, we diagnosed multicentric Castleman's disease (MCD). Light microscopy of kidney biopsy samples revealed various nephropathies, including membranoproliferative glomerulonephritis with crescentic formation and focal segmental sclerosis and tubulointerstitial nephritis. Immunofluorescence and electron microscopy revealed IgG-positive deposits in the subepithelial areas, mesangial areas, and tubular basement membrane. The patient's clinical findings including kidney disease improved after treatment with tocilizumab. MCD is considered to be caused by abnormally high levels of interleukin (IL)-6. Tocilizumab, an IL-6 receptor antagonist, was effective in this patient, indicating that the immune complex-related kidney findings were also related to MCD., Competing Interests: The authors declare no competing financial interests and no conflicts of interest. Figure 1Axillary lymph node biopsy. Hematoxylin and eosin stain showed an increased number of follicles with large hyperplastic germinal centers showing a minor hyaline vascular pattern; however, only partial plasma cell infiltration was seen in the interfollicular region. Original magnification × 200; bar = 30 μm.Figure 2a – d1st kidney biopsy. a: Endocapillary proliferative changes characterized by endothelial cell swelling and hyperplasia (arrow) with mesangiolysis were noted, together with neutrophil and monocyte infiltration. Periodic acid methenamine silver (PAM) staining; original magnification × 400; bar = 50 μm. b: In the four glomeruli with more severe endothelial cell damage, cellular crescent formation (*) was seen, characterized by epithelial cell swelling and hyperplasia with massive hyaline drop degeneration (arrow). Periodic acid-Schiff stain (PAS) staining; original magnification × 400; bar = 50 μm. c: In addition to cellular crescent formation (*), fibrocellular crescent formation (arrow) was also observed. PAM staining; original magnification × 400; bar = 50 μm. d: Focal segmental sclerosis (arrow) was noted. Masson trichrome staining; original magnification × 400; bar = 50 μm.Figure 2e – h1st kidney biopsy. e: Endothelial cell swelling of arterioles (arrow) with lymphocytic infiltration was seen. PAM staining; original magnification × 200; bar = 30 μm. f: Focal inflammatory cell infiltrates in the tubular interstitium and tubulitis were noted. Hematoxylin and eosin staining; original magnification × 200; bar = 30 μm. g: Moderate atherosclerosis was present in the interlobular arteries (large level). Masson trichrome staining; original magnification × 100; bar = 100 μm. h: Hyalinosis of the arterioles was mild. PAS staining; original magnification × 200; bar = 20 μm.Figure 2i1st kidney biopsy. i: Lymph follicle formation was also observed in part of the tubulointerstitium. Original magnification × 200, bar = 50 μm.Figure 2j1st kidney biopsy. j: Immunofluorescence microscopy revealed granular deposits of immunoglobulin G (IgG), IgG1, IgG2, and C3 along the glomerular basement membrane, Bowman’s capsule, and tubular basement membrane. The GBM was partially positive for C1q and positive for IgG4. Original magnification × 100; bar = 20 μm. Figure 2k - n1st kidney biopsy. k: Electron microscopy revealed electron-dense deposits in subepithelial regions (arrow) of the glomerular basement membrane. l: Electron-dense deposits(arrow) in the mesangial area. m: Electron-dense deposits (arrow) on the tubular basement membrane. n: Electron-dense deposits(arrow) on the basement membrane of Bowman’s capsule. Table 1.Renal pathology. Renal biopsy findings1st2ndGlobal sclerosis0/2725/102Segmental sclerosis+–Mesangial matrix expansion±±Mesangial cell proliferation±±AcuteEndocapillary proliferative changes+–Crescent formation+–Interstitial inflammation+++ChronicDuplication of the basement membrane++Spike formation––Tubular atrophy+++Interstitial fibrosis+++Amyloid deposition––IFIgA deposition––IgG deposition (mesangial region)+±IgG deposition (capillary wall)++C3 deposition+–C1q deposition+–The findings from the first and second renal biopsies were recorded. The crescent formation and endothelial cell damage, which were considered acute lesions, showed improvement. Additionally, the deposits of C3 and IgG had also improved. IF = immunofluorescence. Figure 3Clinical course.Figure 42nd kidney biopsy. a+b: The endocapillary proliferative changes seen in the first biopsy were greatly improved. a: PAS staining; original magnification × 400, bar = 20 μm. b: PAM staining; original magnification × 400, bar = 20 μm. c: IgG deposition in IF was reduced in glomeruli; original magnification × 400, bar = 20 μm, d: C3 deposition in IF was reduced in glomeruli; original magnification × 400, bar = 20 μm. e: Subepithelial EDDs (arrow) were greatly reduced in EM, bar = 2.0 μm. f: Inflammatory cell infiltration and fibrosis of the tubulointerstitium improved to 15 – 20% of the cortical area. Masson trichrome staining; original magnification × 100, bar = 500 μm. Table 2.Renal pathology of Castleman’s disease reported to be associated with IL-6. ReferenceDiagnosis by renal pathology or clinical diagnosisTreatment[4]Mesangial matrix proliferation, mesangial cell proliferation, interstitial inflammation, fibroelastosisTocilizumab[5]AA amyloidTocilizumab[6]IgAVMMF + Tac + tocilizumab[7]Mesangial hypercellularity, matrix expansion, GBMs showed segmental double contouringTocilizumab[8]IgAVTocilizumab[10]Tubulointerstitial lymphoplasmatic infiltrates mimicking IgG4-related diseasePSL + tocilizumabThe renal pathology in Castleman’s disease, which has been associated with IL-6, is described. The renal lesions exhibit a variety of characteristics. GBMS = glomerular basement membranes; MMF = mycophenolate mofetil; Tac = tacrolimus; PSL = prednisolone., (© Dustri-Verlag Dr. K. Feistle.)

The electromotility of outer hair cell is considered to be based on voltage-dependent conformational changes in the motor protein prestin. The structure and function of prestin have been increasingly examined in recent years. To obtain further information on prestin, a method to stably obtain prestin as the material for this research is required. This study attempted to construct a stable expression system for prestin on Chinese hamster ovary (CHO) cells and its function was evaluated. His-tagged prestin expression vectors were transfected into CHO cells and high-expression clones were obtained by drug selection and limiting dilution method. The expression and activity of prestin were examined by Western blotting and immunofluorescent experiments and by patch clamping. Clones with high fluorescent intensity tended to exhibit bell-shaped non-linear capacitance, suggesting the active function of prestin in such clones. The stable expression and activity of prestin in CHO cells were confirmed in 7 clones out of twelve clones developed. Although expression levels were 1/60-1/30 of those in OHCs, cell line maintaining the stable expression of functional prestin with His tag will be advantageous for future analyses of its structure and function., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Motoo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Fatty acid binding protein 4 (FABP4) is highly expressed in adipocytes. Lipolysis, caused by an elevated adrenergic input, has been suggested to contribute to elevated serum FABP4 levels in patients with cardiovascular diseases. However, the relationship between the serum FABP4 and efferent sympathetic nerve activity remains poorly understood. Twenty-one healthy subjects (average age, 29.1 years; 15 men) performed an isometric handgrip (HG) exercise at 30% of the maximal voluntary contraction until they were fatigued. The beat-by-beat heart rate (HR), blood pressure (BP), and muscle sympathetic nerve activity (MSNA) were recorded. Blood samples were collected at rest and at the time of peak fatigue. The MSNA, HR, and systolic BP were significantly increased by the HG exercise (all, p < 0.05). MSNA was obtained from 14 patients. The change in the FABP4 on HG exercise was significantly correlated with the change in MSNA (bursts/100 heartbeats) (R = 0.808, p < 0.001) but not with changes in other parameters, which might, in part, reflect an association of efferent sympathetic drive with FABP4. Meanwhile, resting FABP4 levels were not associated with any parameters including MSNA, in healthy individuals. Future studies on patients with elevated sympathetic activity are warranted to examine the relationship further., (© 2024 The Author(s). Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society.)

This case study describes the successful use of ravulizumab in treating a 71-year-old woman with myasthenia gravis experiencing a myasthenic crisis. The patient initially presented with hypernasality and dysphagia; her medical history included untreated, complicated type 1 diabetes. The patient received several treatments approved in Japan for general myasthenia gravis, including immunoadsorption plasmapheresis, tacrolimus, intravenous immunoglobulin, and intravenous methylprednisolone. Despite these treatments, the patient's condition fluctuated, and she ultimately experienced a myasthenic crisis, which required ventilator management. The introduction of ravulizumab, a complement inhibitor targeting the complement protein C5, marked a significant turning point in the patient's treatment. Ravulizumab improved the patient's respiratory function, allowing ventilator weaning and discharge from the hospital. To the best of our knowledge, this case is the first report of successful weaning from mechanical ventilation after treatment with ravulizumab in a patient with a myasthenic crisis. This finding suggests the efficacy of ravulizumab in the management of refractory myasthenia gravis and highlights the potential of novel therapeutic approaches and combination strategies in improving the condition. Future clinical trials are needed to assess the efficacy and safety of ravulizumab in larger, more diverse populations of patients with myasthenia gravis., Competing Interests: Human subjects: Consent for treatment and open access publication was obtained or waived by all participants in this study. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Uchi et al.)

Cancer cells show a dynamic metabolic landscape, requiring a sufficient supply of nucleotides to proliferate. They are highly dependent on de novo purine biosynthetic pathways for their nucleotide requirements. Phosphoribosyl pyrophosphate amidotransferase (PPAT), catalyzing the first step of de novo purine biosynthesis, is highly expressed in various cancers. We observed an increased expression of PPAT in nasopharyngeal carcinoma (NPC). Moreover, our ribonucleic acid sequencing analysis showed high PPAT expression in Epstein-Barr virus-positive NPC, which was supported by in vitro analysis. Through a gene knockdown study, we showed that the suppression of PPAT expression reduced the proliferation and invasion of NPC cells. We also demonstrated the regulation of PPAT by glutamine, a cosubstrate for PPAT. A glutamine antagonist, 6-diazo-5-oxo-L-norleucine, blocked glutamine-mediated induction of PPAT and reduced NPC cell proliferation. Immunohistochemical analysis of PPAT in NPC tissues revealed increased expression of PPAT with disease progression, which was significantly associated with poor prognosis. In summary, this study highlighted the biological function of PPAT in NPC, establishing its potential as a novel prognostic biomarker for aggressive NPC and a promising therapeutic target., (© 2024 The Author(s). Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)

Introduction: Previous randomized controlled trials have not demonstrated the benefits of renal artery stenting with respect to kidney function. However, these trials did not focus on patients with severely impaired kidney function caused by severe bilateral stenosis. Therefore, the efficacy of stenting in such patients remains unclear., Case Presentation: We report 4 cases of successful percutaneous transluminal renal angioplasty (PTRA) with severely impaired kidney function with rapid decline caused by bilateral atherosclerotic stenosis. The catheterization before irreversible parenchymal damages was useful in improving kidney function dramatically in these cases of severe bilateral renal artery stenosis. Furthermore, we examined the clinical characteristics of the 4 cases to identify the potential predictors of PTRA effectiveness. Notably, bilateral renal artery >90% stenosis, elevated plasma renin activity, estimated glomerular filtration rate <15 mL/min/1.73 m2 with an accelerated decline within 6 months before PTRA (>50 mL/min/1.73 m2/6 months), and resistance index (RI) <0.7 were identified as common findings., Conclusion: PTRA should be considered a treatment strategy for patients with these features to preserve kidney function and avoid dialysis therapy., (© 2024 The Author(s). Published by S. Karger AG, Basel.)

Fabry disease, an X-linked lysosomal storage disorder caused by galactosidase α (GLA) gene mutations, exhibits diverse clinical manifestations, and poses significant diagnostic challenges. Early diagnosis and treatment are crucial for improved patient outcomes, pressing the need for reliable biomarkers. In this study, we aimed to identify miRNA candidates as potential biomarkers for Fabry disease using the KingFisher™ automated isolation method and NanoString nCounter® miRNA detection assay. Clinical serum samples were collected from both healthy subjects and Fabry disease patients. RNA extraction from the samples was performed using the KingFisher™ automated isolation method with the MagMAX mirVanaTM kit or manually using the Qiagen miRNeasy kit. The subsequent NanoString nCounter® miRNA detection assay showed consistent performance and no correlation between RNA input concentration and raw count, ensuring reliable and reproducible results. Interestingly, the detection range and highly differential miRNA between the control and disease groups were found to be distinct depending on the isolation method employed. Nevertheless, enrichment analysis of miRNA-targeting genes consistently revealed significant associations with angiogenesis pathways in both isolation methods. Additionally, our investigation into the impact of enzyme replacement therapy on miRNA expression indicated that some differential miRNAs may be sensitive to treatment. Our study provides valuable insights to identify miRNA biomarkers for Fabry disease. While different isolation methods yielded various detection ranges and highly differential miRNAs, the consistent association with angiogenesis pathways suggests their significance in disease progression. These findings lay the groundwork for further investigations and validation studies, ultimately leading to the development of non-invasive and reliable biomarkers to aid in early diagnosis and treatment monitoring for Fabry disease., Competing Interests: JYF, SA, HS, and MGQ are current employees at Takeda and have ownership of Takeda stock. AFP was under employment of Takeda during the project. This work was funded by Takeda Pharmaceuticals International. This commercial affiliation does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2024 Fang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The liver is a complex organ with a highly organized structure in which tight junctions (TJs) play an important role in maintaining their function by regulating barrier properties and cellular polarity. Dysfunction of TJs is associated with liver diseases, including progressive familial intrahepatic cholestasis (PFIC). In this study, we investigated the molecular alterations in a liver-specific ZO-1 and ZO-2 double-knockout (DKO) mouse model, which exhibits features resembling those of PFIC4 patients with mutations in the ZO-2 gene. RNA-seq analysis revealed the upregulation of genes involved in the oxidative stress response, xenobiotic metabolism, and cholesterol metabolism in DKO livers. Conversely, the expression of genes regulated by HNF4α was lower in DKO livers than in the wild-type controls. Furthermore, age-associated analysis elucidated the timing and progression of these pathway changes as well as alterations in molecules related to TJs and apical polarity. Our research uncovered previously unknown implications of ZO-1 and ZO-2 in liver physiology and provides new insights into the molecular pathogenesis of PFIC4 and other tight junction-related liver diseases. These findings contribute to a better understanding of the complex mechanisms underlying liver function and dysfunction and may lead to the development of novel therapeutic strategies for liver diseases associated with tight junction impairment.Key words: tight junctions, ZO-1/ZO-2 knockout mouse, liver, transcriptome analysis, molecular pathological progression.

Optical magnetic dipole (MD) emission predominantly relies on emitters with significant MD transitions, which, however, rarely exist in nature. Here, we propose a strategy to transform electric dipole (ED) emission to a magnetic one by elegantly coupling an ED emitter to a silicon nanoparticle exhibiting a strong MD resonance. This emission mode transformation enables an artificially ideal magnetic dipole source with an MD purity factor of up to 99%. The far-field emission patterns of such artificial MD sources were experimentally measured, which unambiguously resolved their magnetic-type emission origin. This study opens the path to achieving ideal magnetic dipole emission with nonmagnetic emitters, largely extending the availability of magnetic light emitters conventionally limited by nature. Beyond the fundamental significance in science, we anticipate that this study will also facilitate the development of magnetic optical nanosource and enable potential photonic applications relying on magnetic light emission.

Astrocytes are key components of the neurovascular unit. While we have recently identified Olig2+ astrocyte progenitors (ASPs) in the developing mouse dentate gyrus (DG), their molecular signature remains incompletely characterized. Here we demonstrate that Olig2+ ASPs predominantly express brain lipid-binding protein (BLBP), while only a small population of them expresses gfap -GFP. These Olig2+/BLBP+ ASPs co-express the transcription factors Sox3, Sox9 and the proteoglycan NG2 but not Sox10, a marker for oligodendrocyte progenitors (OLPs). Olig2+ ASPs appear from embryonic day 18 (E18) onwards and decline at postnatal day 14 (P14). Consistent with the proliferation of both Olig2+ and NG2+ glial cells after brain injury, intrauterine intermittent hypoxia (IH) led to an increase in Olig2+/NG2+/BLBP+ ASPs in the postnatal DG. IH also promoted both angiogenesis and vascular coupling of Olig2+/NG2+ ASPs. Our data suggest that IH-induced expression of HIF1a increases Olig2+/NG2+/BLBP+ ASPs in a cell non-autonomous manner. Our data also revealed increased vascular coupling of GFAP+ astrocytes following IH, while the number of GFAP+ astrocytes remains unchanged. Given that BLBP, Olig2 and NG2 are expressed in reactive astrocytes, our findings suggest that Olig2+/NG2+/BLBP+ ASPs represent a subtype of reactive astrocyte progenitors. Furthermore, the enhanced vascular coupling of Olig2+/NG2+/BLBP+ ASPs appears to be an adaptive response to hypoxic brain injury. This study provides new insights into the molecular characteristics of Olig2+/NG2+/BLBP+ ASPs and their potential role in the brain's response to hypoxic injury, contributing to our understanding of neurovascular unit dynamics in both development and pathological conditions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Omura, Ogawa, Kawachi, Ogawa, Arai, Takayama, Masui, Kondo, Sugimoto, Shinohara, Takahashi, Maeda and Ohyama.)

Varicella-zoster virus (VZV) infections of the central nervous system can be atypical, particularly in elderly patients. Herein, we describe the case of an 80-year-old male presenting with persistent hiccups and dysphagia without the characteristic rash typically associated with VZV infection. Cerebrospinal fluid multiplex polymerase chain reaction (PCR) allowed rapid identification of VZV, thereby enabling the prompt initiation of antiviral therapy. This case underscores the importance of considering VZV in atypical neurological presentations and highlights the utility of multiplex PCR in diagnosing zoster sine herpes, potentially altering the diagnostic approach for similar cases., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Ethics Committee of Toho University Ohashi Medical Center issued approval exempt. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Uchi et al.)

Objectives: In sepsis treatment, antibiotics are crucial, but overuse risks development of antibiotic resistance. Recent guidelines recommended the use of procalcitonin to guide antibiotic cessation, but solid evidence is insufficient. Recently, concerns were raised that this strategy would increase recurrence. Additionally, optimal protocol or difference from the commonly used C-reactive protein (CRP) are uncertain. We aimed to compare the effectiveness and safety of procalcitonin- or CRP-guided antibiotic cessation strategies with standard of care in sepsis., Data Sources: A systematic search of PubMed, Embase, CENTRAL, Igaku Chuo Zasshi, ClinicalTrials.gov , and World Health Organization International Clinical Trials Platform., Study Selection: Randomized controlled trials involving adults with sepsis in intensive care., Data Extraction: A systematic review with network meta-analyses was performed. The Grading of Recommendations, Assessments, Developments, and Evaluation method was used to assess certainty., Data Synthesis: Eighteen studies involving 5023 participants were included. Procalcitonin-guided and CRP-guided strategies shortened antibiotic treatment (-1.89 days [95% CI, -2.30 to -1.47], -2.56 days [95% CI, -4.21 to -0.91]) with low- to moderate-certainty evidence. In procalcitonin-guided strategies, this benefit was consistent even in subsets with shorter baseline antimicrobial duration (7-10 d) or in Sepsis-3, and more pronounced in procalcitonin cutoff of "0.5 μg/L and 80% reduction." No benefit was observed when monitoring frequency was less than half of the initial 10 days. Procalcitonin-guided strategies lowered mortality (-27 per 1000 participants [95% CI, -45 to -7]) and this was pronounced in Sepsis-3, but CRP-guided strategies led to no difference in mortality. Recurrence did not increase significantly with either strategy (very low to low certainty)., Conclusions: In sepsis, procalcitonin- or CRP-guided antibiotic discontinuation strategies may be beneficial and safe. In particular, the usefulness of procalcitonin guidance for current Sepsis-3, where antimicrobials are used for more than 7 days, was supported. Well-designed studies are needed focusing on monitoring protocol and recurrence., Competing Interests: The authors have disclosed that they do not have any potential conflicts of interest., (Copyright © 2024 by the Society of Critical Care Medicine and Wolters Kluwer Health, Inc. All Rights Reserved.)

Objectives: To investigate the characteristics of outpatients in the pediatric otolaryngology department of a tertiary Japanese hospital before and during the COVID-19 pandemic., Methods: In this retrospective study conducted at a tertiary center in Japan, pediatric patients aged 0-15 years who visited the pediatric otolaryngology department between 2017 and 2022 were included. The number of outpatients in the department was compared between before and during the COVID-19 pandemic (January 2017-December 2022) by year, age, sex, and the disease for which the patient was examined. Additionally, the diseases that most substantially contributed to the change in outpatient visits were evaluated in detail regarding their severity., Results: Among a total of 9219 outpatients, there were no significant differences between 2017-2019 and 2020-2022 (4650 vs. 4569). In contrast, the number of new outpatients decreased by 27.5% (from 1242 to 900) in each of the three years during the COVID-19 pandemic. In particular, the number of patients aged 1-6 years decreased as did that of male patients. Of eight disease categories, oral cavity- and pharyngeal- related diseases decreased the most significantly, by approximately 50%, and the decrease in the number of patients with obstructive sleep apnea (OSA) is thought to have had an impact on the decrease in the number of patients. Furthermore, in cases in which polysomnography testing was performed for OSA, the proportion of patients with moderate and severe categories showed a significant decrease., Conclusions: The number of new pediatric ORL patients, particularly those with severe OSA, reduced during the COVID 19 pandemic. These results may provide indications about the characteristics of OSA incidence and exacerbations., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

mRNA incorporated in lipid nanoparticles (LNPs) became a new class of vaccine modality for induction of immunity against COVID-19 and ushered in a new era in vaccine development. Here, we report a novel, easy-to-execute, and cost effective engineered extracellular vesicles (EVs)-based combined mRNA and protein vaccine platform (EV X-M+P vaccine) and explore its utility in proof-of-concept immunity studies in the settings of cancer and infectious disease. As a first example, we engineered EVs, natural nanoparticle carriers shed by all cells, to contain ovalbumin mRNA and protein (EV OvaM+P vaccine) to serve as cancer vaccine against ovalbumin-expressing melanoma tumors. EV OvaM+P administration to mice with established melanoma tumors resulted in tumor regression associated with effective humoral and adaptive immune responses. As a second example, we generated engineered EVs that contain Spike (S) mRNA and protein to serve as a combined mRNA and protein vaccine (EV SpikeM+P vaccine) against SARS-CoV-2 infection. EV SpikeM+P vaccine administration in mice and baboons elicited robust production of neutralizing IgG antibodies against RBD (receptor binding domain) of S protein and S protein specific T cell responses. Our proof-of-concept study describes a new platform with an ability for rapid development of combination mRNA and protein vaccines employing EVs for deployment against cancer and other diseases., Competing Interests: Declaration of competing interest MD Anderson Cancer Center and RK have filed patent applications and patents issued in the area of exosome biology, and some of them are licensed to PranaX, Inc. for non-cancer related use. MD Anderson and RK hold equity in PranaX and RK serves as an advisor on non-cancer related matters., (Copyright © 2024. Published by Elsevier B.V.)

[Purpose] This study investigated the test-retest reliability of the ten-repetition maximum leg press (LP) and chest press (CP) tests in untrained young males and females. The secondary aim was to examine the test-retest reliability of the 10RM test according to gender. [Participants and Methods] All participants underwent the ten-repetition maximum test for the LP and CP across three sessions separated by 2 to 6 days of recovery: the first session was familiarization, the second for the initial test, and the third for the retest. To evaluate test-retest reliability, we calculated the intraclass correlation coefficient, standard error of measurement, and minimal detectable change for both the LP and CP. These measures were analyzed separately for each gender to assess gender-related differences. [Results] High intraclass correlation coefficient index was obtained for both LP and CP for both genderes. The standard errors of measurement and minimal detectable changes were consistent with previously reported values. However, females showed more variability than males, particularly in the LP. [Conclusion] The ten-repetition maximum test for LP and CP is a reliable method for measuring strength in untrained young males and females. However, it should be noted that the margin of error may be higher among female participants compared to their male counterparts., Competing Interests: None., (2024©by the Society of Physical Therapy Science. Published by IPEC Inc.)

Objective: To evaluate the efficacy and safety of first-line biological disease-modifying antirheumatic drugs (bDMARDs) in patients with rheumatoid arthritis (RA) with chronic kidney disease (CKD), including those undergoing haemodialysis (HD)., Methods: This retrospective cohort study included 425 patients with RA prescribed their first bDMARDs at two hospitals from 2004 to 2021. Patients were categorised by kidney function and bDMARD modality (TNFα inhibitors (TNFαis), interleukin-6 inhibitors (IL-6is), cytotoxic T-lymphocyte antigen-4 immunoglobulin (CTLA4-Ig)). The primary outcome was the 36-month drug retention rate, with secondary outcomes including changes in Disease Activity Score 28 (DAS28)-C reactive protein (CRP)/erythrocyte sedimentation rate (ESR), prednisolone dosage and reasons for discontinuation., Results: The 36-month drug retention rates by estimated glomerular filtration rate (eGFR) (≥60, 30-60, <30 mL/min/1.73 m 2 ) were as follows: all bDMARDs (45.2%, 32.0%, 41.4%), TNFαis (45.3%, 28.2%, 34.0%), IL-6is (47.4%, 66.7%, 71.4%) and CTLA-4Ig (50.0%, 31.3%, 33.3%). Even in groups with lower kidney function, the drug retention rate of bDMARDs was generally maintained. However, the retention rate of TNFαis was significantly lower in patients with eGFR <30 mL/min/1.73 m 2 . IL-6is showed the highest retention rate and the lowest discontinuation rate due to ineffectiveness in this group (HR 0.11, 95% CI 0.02 to 0.85, p=0.03). All bDMARDs improved DAS28-CRP/ESR and reduced prednisolone dosage across all groups., Conclusion: bDMARDs demonstrated effective and safe profiles in patients with RA with CKD, even among patients on HD. In particular, IL-6is had a significantly higher drug retention rate in patients with an eGFR of <30 mL/min/1.73 m 2 and fewer discontinuations due to ineffectiveness. IL-6is were more efficacious as monotherapy compared with the other bDMARDs., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ on behalf of EULAR.)

Hepatic glucose metabolism serves dual purposes: maintaining glucose homeostasis and converting glucose into energy sources; however, the underlying mechanisms are unclear. We quantitatively measured liver metabolites, gene expression, and phosphorylated insulin signaling molecules in mice orally administered varying doses of glucose, and constructed a transomic network. Rapid phosphorylation of insulin signaling molecules in response to glucose intake was observed, in contrast to the more gradual changes in gene expression. Glycolytic and gluconeogenic metabolites and expression of genes involved in glucose metabolism including glucose-6-phosphate, G6pc, and Pck1, demonstrated high glucose dose sensitivity. Whereas, glucokinase expression and glycogen accumulation showed low glucose dose sensitivity. During the early phase after glucose intake, metabolic flux was geared towards glucose homeostasis regardless of the glucose dose but shifted towards energy conversion during the late phase at higher glucose doses. Our research provides a comprehensive view of time- and dose-dependent selective glucose metabolism., (© 2024. The Author(s).)

Motivation: Visualization and analysis of biological networks play crucial roles in understanding living systems. Biological networks include diverse types, from gene regulatory networks and protein-protein interactions to metabolic networks. Metabolic networks include substrates, products, and enzymes, which are regulated by allosteric mechanisms and gene expression. However, the analysis of these diverse omics types is challenging due to the diversity of databases and the complexity of network analysis., Results: We developed iTraNet, a web application that visualizes and analyses trans-omics networks involving four types of networks: gene regulatory networks, protein-protein interactions, metabolic networks, and metabolite exchange networks. Using iTraNet, we found that in wild-type mice, hub molecules within the network tended to respond to glucose administration, whereas in ob/ob mice, this tendency disappeared. With its ability to facilitate network analysis, we anticipate that iTraNet will help researchers gain insights into living systems., Availability and Implementation: iTraNet is available at https://itranet.streamlit.app/., Competing Interests: None declared., (© The Author(s) 2024. Published by Oxford University Press.)

Background: Stanford type B-acute aortic dissection (type B-AAD) is often life-threatening without invasive surgery. Multilineage-differentiating stress enduring cell (Muse cells), which comprise several percent of mesenchymal stem cells (MSCs), are endogenous pluripotent-like stem cells that selectively home to damaged tissue and replace damaged/apoptotic cells by in-vivo differentiation., Methods: Mortality, aortic diameter expansion, cell localization, cell differentiation, and inflammation of the dissected aorta were evaluated in type B-AAD model mice intravenously injected with human-Muse cells, -elastin-knockdown (KD)-Muse cells, -human leukocyte antigen-G (HLA-G)-KD-Muse cells, or MSCs, all without immunosuppressant., Results: Here, we show the Muse (50,000 cells) group has a lower incidence of aortic rupture and mortality of AAD compared with the MSC-50K (50,000 human-MSCs) and vehicle groups. Spectrum computed tomography in-vivo dynamics and 3-dimensional histologic analyses demonstrate that Muse cells more effectively home to the AAD tissue and survive for 8 weeks in the Muse group than in the MSC-750K (750,000 human-MSCs containing 50,000 Muse cells) group. Homing of Muse cells is impeded in the HLA-G-KD-Muse (50,000 cells) group. Differentiation of homed Muse cells into CD31(+) and alpha-smooth muscle actin (+) cells, production and reorganization of elastic fibers in the AAD tissue, and suppression of diameter expansion are greater in the Muse group than in the MSC-750K and elastin-KD-Muse (50,000 cells) groups., Conclusions: Intravenously administered Muse cells reconstruct the dissected aorta and improve mortality and diameter enlargement rates. Moreover, small doses of purified Muse cells are more effective than large doses of MSCs. HLA-G is suggested to contribute to the successful survival and homing of Muse cells., (© 2024. The Author(s).)

Myasthenia gravis (MG) patients often require long-term glucocorticoid therapy, which may affect bone health. This study aimed to assess long-term changes in bone mineral density (BMD), evaluate osteoporotic fracture incidence, and examine the relationship between MG-specific factors and bone health outcomes over a 10-year period. This single-center, prospective cohort study included 28 MG patients. BMD, T-scores, Z-scores, and bone turnover markers were measured at baseline. FRAX ® scores were calculated and adjusted for glucocorticoid dose. Fracture occurrence was monitored for over 10 years. Five (17.9%) patients experienced major osteoporotic fractures during follow-up. The fracture group had significantly lower baseline BMD and T-scores than the no-fracture group. Baseline FRAX ® scores for major osteoporotic fracture risk were significantly higher in the fracture group (median 19.0% vs. 5.7%, p = 0.001). The fracture group progressed from osteopenia at baseline to osteoporosis by the end of this study. This study highlights the importance of early and regular bone health assessments in MG patients, particularly those receiving long-term glucocorticoid therapy. The FRAX ® tool may be valuable for fracture risk stratification in this population. These findings can inform clinical practice and improve long-term management strategies for MG patients who are at risk of osteoporotic fractures.

Carcinomas are associated with metastasis to specific organs while sparing others. Breast cancer presents with lung metastasis but rarely kidney metastasis. Using this difference as an example, we queried the mechanism(s) behind the proclivity for organ-specific metastasis. We used spontaneous and implant models of metastatic mammary carcinoma coupled with inflammatory tissue fibrosis, single-cell sequencing analyses and functional studies to unravel the causal determinants of organ-specific metastasis. Here we show that lung metastasis is facilitated by angiopoietin 2 (Ang2)-mediated suppression of lung-specific endothelial tight junction protein Claudin 5, which is augmented by the inflammatory fibrotic microenvironment and prevented by anti-Ang2 blocking antibodies, while kidney metastasis is prevented by non-Ang2-responsive Claudins 2 and 10. Suppression of Claudins 2 and 10 was sufficient to induce the emergence of kidney metastasis. This study illustrates the influence of organ-specific vascular heterogeneity in determining organotropic metastasis, independent of cancer cell-intrinsic mechanisms., (© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.)

This study investigates the promotion of sodium chlorate (NaClO 3 ) crystallization through optical trapping, enhanced by the addition of gold nanoparticles (AuNPs) and silicon nanoparticles (SiNPs). Using a focused laser beam at the air-solution interface of a saturated NaClO 3 solution with AuNPs or SiNPs, the aggregates of these particles were formed at the laser focus, the nucleation and growth of metastable NaClO 3 (m-NaClO 3 ) crystals were induced. Continued laser irradiation caused these m-NaClO 3 crystals to undergo repeated cycles of growth and dissolution, eventually transitioning to a stable crystal form. Our comparative analysis showed that AuNPs, due to their significant heating due to higher photon absorption efficiency, caused more pronounced size fluctuations in m-NaClO 3 crystals compared to the stable behavior observed with SiNPs. Interestingly, the maximum diameter of the m-NaClO 3 crystals that appeared during the size fluctuation step was consistent, regardless of nanoparticle type, concentration, or size. The crystallization process was also promoted by using polystyrene nanoparticles, which have minimal heating and electric field enhancement, suggesting that the reduction in activation energy for nucleation at the particle surface is a key factor. These findings provide critical insights into the mechanisms of laser-induced crystallization, emphasizing the roles of plasmonic heating, particle surfaces, and optical forces., (© 2024. The Author(s).)

We investigated the presence and functionality of the carbon storage regulator (Csr) system in Aeromonas salmonicida SWSY-1.411. CsrA, an RNA-binding protein, shared 89% amino acid sequence identity with Escherichia coli CsrA. CsrB/C sRNAs exhibited a typical stem-loop structure, with more GGA motifs, which bind CsrA, than E. coli. CsrD had limited sequence identity with E. coli CsrD; however, it contained the conserved GGDEF and EAL domains. Functional analysis in E. coli demonstrated that the Csr system of A. salmonicida influences glycogen biosynthesis, biofilm formation, motility, and stability of both CsrB and CsrC sRNAs. These findings suggest that in A. salmonicida, the Csr system affects phenotypes like its E. coli counterpart. In A. salmonicida, defects in csr homologs affected biofilm formation, motility, and chitinase production. However, glycogen accumulation and protease production were unaffected. The expression of flagellar-related genes and chitinase genes was suppressed in the csrA-deficient A. salmonicida. Northern blot analysis indicated the stabilization of CsrB and CsrC in the csrD-deficient A. salmonicida. Similar to that in E. coli, the Csr system in A. salmonicida comprises the RNA-binding protein CsrA, the sRNAs CsrB and CsrC, and the sRNA decay factor CsrD. This study underscores the conservation and functionality of the Csr system and raises questions about its regulatory targets and mechanisms in A. salmonicida.

Control of circularly polarized light (CPL) is important for next-generation optical communications as well as for investigating the optical properties of materials. In this study, we explore dielectric-sphere oligomers for chiral nanoantenna applications, leveraging the cathodoluminescence (CL) technique, which employs accelerated free electrons for excitation and allows mapping the optical response on the nanoscale. For a certain particle-dimers configuration, one of the spheres becomes responsible for the left-handed circular polarization of the emitted light, while right-handed circular polarization is selectively yielded when the other sphere is excited by the electron beam. Similar patterns are also observed in trimers. These phenomena are understood in terms of optical coupling between the electric and magnetic modes hosted by the dielectric spheres. Our research not only expands the understanding of CPL generation mechanisms in dielectric-sphere oligomer antennas but also underscores the potential of such structures in optical applications. We further highlight the utility of CL as a powerful analytical tool for investigating the optical properties of nanoscale structures as well as the potential of electron beams for light generation with switchable CPL parities., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

The mechanism of oxidation of phenols by tetrahedral copper(II)-halide complexes was investigated to demonstrate that phenols with an electron-withdrawing substituent are oxidized via a proton-transfer/electron-transfer (PTET) mechanism, whereas phenols with an electron-donating substituent involve a concerted proton/electron transfer (CPET) mechanism. The importance of the tetrahedral geometry of the metal centre as well as the effects of the halide ligands of the substrates were explored.

A tumor microenvironment (TME)-responsive nanoprobe composed of a fluorescent dye-decorated silicon (Si) nanosphere core and a thin MnO 2 shell is proposed for simple and intelligent detection of cancer cells. The Si nanosphere core with diameters of 100-200 nm provides environment-independent Mie scattering imaging, while, simultaneously, the MnO 2 shell provides the capability to switch the on/off state of the dye fluorescence reacted to the glutathione (GSH) and/or H 2 O 2 levels in a cancer cell. Si-MnO 2 core-shell nanosphere probes are fabricated in a solution-based process from crystalline Si nanosphere cores. The fluorescence switching under exposure to GSH is demonstrated, and the mechanism is discussed based on detailed optical characterizations including single-particle spectroscopy. Different types of human cells are incubated with the nanoprobes, and a proof of concept experiment is performed. From the combination of the robust scattering images and GSH- and H 2 O 2 -sensitive fluorescence images, the feasibility of cancer cell detection by the multimodal nanoprobes is demonstrated.

Developing a cost-effective method for separating and concentrating tritium water (HTO) from light water (H 2 O) without consuming additional energy is crucial for achieving reliable and safe nuclear fission and fusion energy technologies. However, this presents a significant challenge because of the difficulties in obtaining basic information, such as the chemical and physical properties of HTO molecules. Here, we investigate the isotope exchange reaction (IER) between HTO molecules in H 2 O solution and H 2 O vapor in the atmosphere. The reduction and purification rates of HTO-containing water were measured by varying the system conditions, such as temperature (20-50 °C) and humidity (50 %-90 %), under an equilibrium state between the liquid phase (water) and vapor phase (air). Our findings indicate that the concentration of HTO in the solution can be significantly reduced by increasing H 2 O vapor in the atmosphere. This result can be quantitatively explained by considering the entropy of mixing between the solution and vapor phases. The results obtained here provide both basic understanding on the exchange process between liquid- and vapor-water molecules and a passive technology for treating HTO-containing water., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Inflammation and tissue damage associated with pancreatitis can precede or occur concurrently with pancreatic ductal adenocarcinoma (PDAC). We demonstrate that in PDAC coupled with pancreatitis (ptPDAC), antigen-presenting type I conventional dendritic cells (cDC1s) are specifically activated. Immune checkpoint blockade therapy (iCBT) leads to cytotoxic CD8 + T cell activation and elimination of ptPDAC with restoration of life span even upon PDAC rechallenge. Using PDAC antigen-loaded cDC1s as a vaccine, immunotherapy-resistant PDAC was rendered sensitive to iCBT with elimination of tumors. cDC1 vaccination coupled with iCBT identified specific CDR3 sequences in the tumor-infiltrating CD8 + T cells with potential therapeutic importance. This study identifies a fundamental difference in the immune microenvironment in PDAC concurrent with, or without, pancreatitis and provides a rationale for combining cDC1 vaccination with iCBT as a potential treatment option.

Nanotechnology has revolutionized the fabrication of hybrid species with tailored functionalities. A milestone in this field is the deoxyribonucleic acid (DNA) conjugation of nanoparticles, introduced almost 30 years ago, which typically exploits the affinity between thiol groups and metallic surfaces. Over the last decades, developments in colloidal research have enabled the synthesis of an assortment of nonmetallic structures, such as high-index dielectric nanoparticles, with unique properties not previously accessible with traditional metallic nanoparticles. However, to stabilize, integrate, and provide further functionality to nonmetallic nanoparticles, reliable techniques for their functionalization with DNA will be crucial. Here, we combine well-established dibenzylcyclooctyne-azide click-chemistry with a simple freeze-thaw method to achieve the functionalization of silica and silicon nanoparticles, which form exceptionally stable colloids with a high DNA surface density of ∼0.2 molecules/nm 2 . Furthermore, we demonstrate that these functionalized colloids can be self-assembled into high-index dielectric dimers with a yield of over 50% via the use of DNA origami. Finally, we extend this method to functionalize other important nanomaterials, including oxides, polymers, core-shell, and metal nanostructures. Our results indicate that the method presented herein serves as a crucial complement to conventional thiol functionalization chemistry and thus greatly expands the toolbox of DNA-functionalized nanoparticles currently available.

Introduction: Myasthenia gravis (MG), an immune disorder affecting nerve-muscle transmission, often necessitates tailored therapies to alleviate longitudinal symptom fluctuations. Here, we aimed to examine and compare the treatment cycle intervals and efficacy of efgartigimod in four patients. This case series mainly offers insights into personalized treatment cycle intervals and the efficacy of efgartigimod for patients with MG in our facility in Japan., Methods: We retrospectively analyzed four patients with MG (2 patients with early-onset, 1 with late-onset, and 1 with seronegative MG, mainly managed with oral immunosuppressants as prior treatments) who completed four or more cycles of efgartigimod treatment from January 2022 to September 2023. We focused on changes in serum immunoglobulin (IgG) level, acetylcholine receptor antibody (AChR-Ab) titer, and quantitative MG (QMG) score., Results: Efgartigimod, administered at a median of 5.0 [IQR 5.0, 7.5] weeks between cycles, led to decreased serum IgG levels in all patients and reduced AChR-Ab titers in seropositive patients. All patients showed sustained MG symptom improvement, with considerably reduced QMG scores before efgartigimod treatment. None of the patients required rescue medications or developed treatment-related adverse events., Conclusions: Customized efgartigimod administration intervals effectively enhanced clinical outcomes in patients with MG without notable symptom fluctuations, demonstrating the benefits of individualized treatment approaches and validating the safety of efgartigimod during the study period.

Background: Myasthenia gravis (MG) is an immune disorder that causes muscle weakness with an increasing prevalence, particularly among the elderly in Japan. Glucocorticoid treatment for MG is problematic for bone health because of reduced bone density and increased fracture risk. The fracture risk assessment tool (FRAX®) can estimate fracture risk, but its applicability in patients with MG remains uncertain., Methods: A prospective cohort study was conducted on 54 patients with MG between April and July 2012. Bone mineral density (BMD) was measured, and FRAX® scores were calculated with and without BMD. We also adjusted FRAX® scores based on glucocorticoid dosage. Patients were monitored for major osteoporotic fractures (MOF) until June 2022. Statistical analyses included Kaplan-Meier curves and Cox proportional hazards models., Results: The study group included 12 men and 42 women with a mean age of 62 years. Higher FRAX® scores correlated with increased fracture risk, particularly in the hip and lumbar regions. The 10-year fracture-free rate was significantly lower in the high-FRAX® score group. The FRAX® score using BMD is a significant predictor of MOF risk. The hazard ratio for FRAX® scores was 1.17 (95% CI 1.10-1.26)., Conclusion: We demonstrated the effectiveness of the FRAX® tool in assessing fracture risk among patients with MG. High FRAX® scores correlated with increased fracture risk, emphasizing its importance. These findings support the incorporation of FRAX® assessment into clinical management to enhance patient care and outcomes. However, the small sample size and observational nature suggest a need for further research., Competing Interests: Declaration of competing interest None declared., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Pituitary tumors (PTs) represent about 10% of all intracranial tumors, and most are benign. However, some PTs exhibit continued growth despite multimodal therapies. Although temozolomide (TMZ), an alkylating chemotherapeutic agent, is a first-line medical treatment for aggressive PTs, some PTs are resistant to TMZ. Existing literature indicated the involvement of autophagy in cell growth in several types of tumors, including PTs, and autophagy inhibitors have anti-tumor effects. In this study, the expression of several autophagy-inducible genes, including Atg3, Beclin1, Map1lc3A, Map1lc3b, Ulk1, Wipi2, and Tfe3 in two PT cell lines, the mouse corticotroph AtT-20 cells and the rat mammosomatotroph GH4 cells were identified. Down regulation of Tfe3, a master switch of basal autophagy, using RNA interference, suppressed cell proliferation in AtT-20 cells, suggesting basal autophagy contributes to the maintenance of cellular functions in PT cells. Expectedly, treatment with bafilomycin A1, an autophagy inhibitor, suppressed cell proliferation, increased the cleavage of PARP1, and reduced ACTH production in AtT-20 cells. Treatment with two additional autophagy inhibitors, chloroquine (CQ) and monensin, demonstrated similar effects on cell proliferation, apoptosis, and ACTH production in AtT-20 cells. Also, treatment with CQ suppressed cell proliferation and growth hormone production in GH4 cells. Moreover, the combination of CQ and TMZ had an additive effect on the inhibition of cell proliferation in AtT-20 and GH4 cells. The additive effect of anti-cancer drugs such as CQ alone or in combination with TMZ may represent a novel therapeutic approach for PTs, in particular tumors with resistance to TMZ., Competing Interests: Declaration of competing interest The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Evaluating the heterogeneity of extracellular vesicles (EVs) is crucial for unraveling their complex actions and biodistribution. Here, we identify consistent architectural heterogeneity of EVs using cryogenic transmission electron microscopy (cryo-TEM), which has an inherent ability to image biological samples without harsh labeling methods while preserving their native conformation. Imaging EVs isolated using different methodologies from distinct sources, such as cancer cells, normal cells, immortalized cells, and body fluids, we identify a structural atlas of their dominantly consistent shapes. We identify EV architectural attributes by utilizing a segmentation neural network model. In total, 7,576 individual EVs were imaged and quantified by our computational pipeline. Across all 7,576 independent EVs, the average eccentricity was 0.5366 ± 0.2, and the average equivalent diameter was 132.43 ± 67 nm. The architectural heterogeneity was consistent across all sources of EVs, independent of purification techniques, and compromised of single spherical, rod-like or tubular, and double shapes. This study will serve as a reference foundation for high-resolution images of EVs and offer insights into their potential biological impact.

Maternal immunoglobulin (Ig)G is present in breast milk and has been shown to contribute to the development of the immune system in infants. In contrast, maternal IgG has no known effect on early childhood brain development. We found maternal IgG immunoreactivity in microglia, which are resident macrophages of the central nervous system of the pup brain, peaking at postnatal one week. Strong IgG immunoreactivity was observed in microglia in the corpus callosum and cerebellar white matter. IgG stimulation of primary cultured microglia activated the type I interferon feedback loop by Syk. Analysis of neonatal Fc receptor knockout (FcRn KO) mice that could not take up IgG from their mothers revealed abnormalities in the proliferation and/or survival of microglia, oligodendrocytes, and some types of interneurons. Moreover, FcRn KO mice also exhibited abnormalities in social behavior and lower locomotor activity in their home cages. Thus, changes in the mother-derived IgG levels affect brain development in offsprings., (© 2024. The Author(s).)

Introduction: This study aimed to analyze the clinical course of TAFRO syndrome in patients through extended follow-up, focusing on recurrent cases and long-term remission., Methods: This was a retrospective case series study. We assessed the clinical course of patients diagnosed with TAFRO syndrome between January 2012 and September 2022 at Toranomon Hospital or Toranomon Hospital Kajigaya, excluding those patients who died during the initial hospitalization., Results: Twelve patients were included. Baseline characteristics, laboratory findings, treatment modalities, and outcomes were assessed. During the median follow-up period of 1474 days, two patients experienced recurrence following a reduction in tocilizumab (TCZ) dose, whereas two achieved remission for >400 days without TCZ treatment. The remaining eight patients maintained remission under the continued TCZ therapy. Recurrence diagnosis was complicated by the non-simultaneous presentation of the five manifestations of TAFRO syndrome. The patients who experienced recurrence showed milder manifestations and faster recovery than the initial onset. Glomerular endotheliopathy was evident in kidney biopsies during recurrence, which was similar to the initial presentation. In a case where only inflammation preceded other manifestation, a kidney biopsy was pivotal in distinguishing TAFRO syndrome relapse from other inflammatory conditions such as infection. Pretreatment serum IL-6 levels were within the reference range only in patients who experienced long-term remission without TCZ treatment., Conclusions: This is the first study to perform kidney biopsies on recurrent TAFRO cases, highlighting recurrence after TCZ dosage reduction, non-simultaneous manifestation of symptoms, the utility of kidney biopsies in recurrence diagnosis, and potential non-IL-6 pathogenesis factors. Pretreatment serum IL-6 levels may help identify patients suitable for maintenance therapy without TCZ. Further investigation is warranted to identify stratified treatment approaches based on individual etiologic factors., Competing Interests: The authors declare that they have no competing or conflicts of interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of the ERA.)

Background: Asking questions is common in conversations, and while asking questions, we need to listen carefully to what others say and consider the perspective our questions adopt. However, difficulties persist in verifying the effect of asking questions on older adults' cognitive function due to the lack of a standardized system for conducting experiments at participants' homes., Objective: This study examined the intervention effect of cognitive training moderated by robots on healthy older adults. A focus on the feasibility of the intervention at participants' homes was also maintained. Feasibility was evaluated by considering both the dropout rate during the intervention and the number of questions posed to each participant during the experiment., Methods: We conducted a randomized controlled trial with 81 adults older than 65 years. Participants were recruited through postal invitations and then randomized into 2 groups. The intervention group (n=40) received sessions where participants listened to photo-integrated stories and posed questions to the robots. The control group (n=41) received sessions where participants listened to photo-integrated stories and only thanked the robots for confirming participation. The participants participated in 12 dialogue sessions for 2-3 weeks. Scores of global cognitive functioning tests, recall tests, and verbal fluency tasks measured before and after the intervention were compared between the 2 groups., Results: There was no significant intervention effect on the Telephone Interview for Cognitive Status-Japanese scores, recall tests, and verbal fluency tasks. Additionally, our study successfully concluded with no participant dropouts at follow-up, confirming the feasibility of our approach., Conclusions: There was no statistically significant evidence indicating intervention benefits for cognitive functioning. Although the feasibility of home-based interventions was demonstrated, we identified areas for improvement in the future, such as setting up more efficient session themes. Further research is required to identify the effectiveness of an improved cognitive intervention involving the act of asking questions., (© Seiki Tokunaga, Takuya Sekiguchi, Kumi Watanabe Miura, Hikaru Sugimoto, Masato S Abe, Kazuhiro Tamura, Taishiro Kishimoto, Takashi Kudo, Mihoko Otake-Matsuura. Originally published in JMIR Aging (https://aging.jmir.org).)

Surgery is the most effective treatment for early-stage lung cancer; however, it poses a higher physical burden than other treatment options. Therefore, understanding the perioperative course of patients is important. Using the Short Form Health Survey 36, we prospectively measured the physical quality of life of patients who underwent anatomical pulmonary resection for non-small cell lung cancer at Shonan Kamakura General Hospital, Kanagawa, Japan (n = 87). In the preoperative setting, patients who had lower performance status and lived alone had significantly worse physical quality of life scores on multivariate analysis (regression coefficient (95% confidence interval), -9.37 (-13.43--5.32) and -10.22 (-13.74--7.40), respectively, p < 0.0001 for both). At 6 months postoperatively, patients who stopped smoking within 1 year preoperatively (stopped smoking within 1 year vs. remote or never smokers, 41.0 ± 10.5 vs. 48.6 ± 7.2, p = 0.002), had lower performance status (0 vs. 1-2, 49.3 ± 6.6 vs. 38.6 ± 9.6, p < 0.0001), lived alone (living alone vs. living with somebody, 41.6 ± 9.7 vs. 48.1 ± 7.9, p = 0.021), and had higher comorbid burden (Charlson comorbidity index <3 vs. ≥3, 48.2 ± 6.9 vs. 39.1 ± 14.7, p = 0.003) had significantly worse physical quality of life scores on univariate analysis. More recent smoking (regression coefficient (95% confidence interval), -4.90 (-8.78-1.0), p = 0.014), lower performance status (8.90 (5.10-12.70), p < 0.0001), living alone (5.76 (1.39-10.13), p = 0.01), and higher comorbid burden (-6.94 (-11.78--2.10), p = 0.006) were significant independent predictors of worse postoperative physical quality of life on multivariate analysis. Therefore, patients with these conditions might need additional support to maintain their physical condition after anatomical lung cancer surgery.

Drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome is a severe drug eruption that causes multiple organ damage. The renal impairment in these patients usually improves with immunosuppressants, but subsequent infections often develop. We herein report a rare case of DRESS syndrome leading to hemodialysis and multiple infections with Pneumocystis pneumonia, cytomegalovirus and Aspergillus despite the administration of low-dose prednisolone. We also present a literature review of cases requiring dialysis after DRESS syndrome. In patients with chronic kidney disease, it is important to be alert for not only the development of DRESS syndrome but also subsequent infections.

Context: Sodium-glucose cotransporter 2 (SGLT2) inhibitors lower blood glucose levels by promoting urinary glucose excretion, but their overall effects on hormonal and metabolic status remain unclear., Objective: We here investigated the roles of insulin and glucagon in the regulation of glycemia in individuals treated with an SGLT2 inhibitor using mathematical model analysis., Methods: Hyperinsulinemic-euglycemic clamp and oral glucose tolerance tests were performed in 68 individuals with type 2 diabetes treated with the SGLT2 inhibitor dapagliflozin. Data previously obtained from such tests in 120 subjects with various levels of glucose tolerance and not treated with an SGLT2 inhibitor were examined as a control. Mathematical models of the feedback loops connecting glucose and insulin (GI model) or glucose, insulin, and glucagon (GIG model) were generated., Results: Analysis with the GI model revealed that the disposition index /clearance , which is defined as the product of insulin sensitivity and insulin secretion divided by the square of insulin clearance and represents the glucose-handling ability of insulin, was significantly correlated with glycemia in subjects not taking an SGLT2 inhibitor but not in those taking dapagliflozin. Analysis with the GIG model revealed that a metric defined as the product of glucagon sensitivity and glucagon secretion divided by glucagon clearance (designated production index /clearance ) was significantly correlated with blood glucose level in subjects treated with dapagliflozin., Conclusion: Treatment with an SGLT2 inhibitor alters the relation between insulin effect and blood glucose concentration, and glucagon effect may account for variation in glycemia among individuals treated with such drugs., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Endocrine Society.)