This study aimed to investigate the efficacy of dapagliflozin administration time using light-dark cycle-disrupted mice. Five-week-old male C57BL/6J mice were fed a control diet or high-fat diet (HFD) for 8-weeks under a disrupted light-dark cycle. During the final 2-weeks of the study, the mice were administered dapagliflozin at a fixed time (8:00) or 2-h after the light phase, respectively. An olive oil-ethanol emulsion was used as the vehicle. At the end of the experiment, the mice were euthanized after an 18-hour fasting period, and plasma and tissue samples (epididymal white adipose tissue, liver, and kidney) were collected. Dapagliflozin administration significantly reduced the HFD-induced body weight gain in both treatment groups. However, the effect of dapagliflozin on body weight reduction was stronger at a fixed time than 2-h after the disrupted light phase. In addition, the administration of dapagliflozin at a fixed time significantly decreased HFD-induced affects such as hyperglycemia and hyperinsulinemia. Although dapagliflozin administration 2-h after the light phase tended to be effective, the levels were lower than those at the fixed time. Our findings suggest that the time to take medicines such as dapagliflozin in shift workers should be fixed at a regular time rather than after waking up.

Gastric cancer (GC) is defined as the primary epithelial malignancy derived from the stomach. This study aims to investigate the predictive and prognostic values of mitotic spindle positioning (MISP) expression in patients with GC, providing new strategies for GC diagnosis and prognosis. MISP expression in GC tissues and adjacent tissues from 120 GC patients was detected. The correlation between pathological characteristics and MISP expression was analyzed. Pathological conditions of patients after 5 years of postoperative treatment were recorded. The correlation between MISP expression and postoperative outcomes of GC patients was analyzed using the Kaplan-Meier method. Cox proportional hazards model was used to analyze independent risk factors affecting the 5-year survival rate of GC patients. MISP expression in GC tissues was higher than in adjacent tissues. MISP expression was correlated with differentiation degree, invasion, lymph node metastasis, and Lauren classification. Patients in the high MISP expression group tended to have poorer postoperative outcomes. High MISP expression was an independent risk factor for the 5-year survival rate of GC patients. In conclusion, high expression of MISP in GC patients indicates a higher possibility of adverse prognosis and is an independent risk factor affecting the post-operative survival rate of GC patients.

Aging promotes mild but progressive decline of motor behaviors and cognitive functions, related to loss of plasticity. Recovery of neural plasticity may become a possible clinical treatment against aging. Perineuronal net (PNN) is the extracellular matrix surrounding cell bodies of neurons, and is known to affect plasticity and survival of neurons. The basal ganglia are motor-related regions of the brain, and earlier reports revealed that neurons in the output nuclei of the basal ganglia form PNN. However, the detailed characteristics are not fully clarified yet. We found intensive PNN expression in the entopeduncular nucleus (EP) and substantia nigra pars reticulata (SNr). PNN surrounded parvalbumin-expressing large axon terminals in the EP and SNr. The proportion of PNN-forming neurons was lower in young mice, and gradually increased with age. The fluorescent intensity of PNN showed similar changes in the EP and SNr. We also showed that the motor ability examined by the rotarod test correlated with PNN expression in the SNr. It may provide the possibility of PNN modulation to improve plasticity and quality of behaviors in aged animals.

Transient receptor potential vanilloid 2 (TRPV2) is a non-selective cation channel activated by mechanical stimuli and temperatures above 52°C. Although we have previously reported that TRPV2 regulates non-shivering thermogenesis through facilitating the expression of genes related to thermogenesis, how TRPV2 activity is regulated in brown adipocytes under physiological conditions remains unclear. Recently, methionine oxidation was reported to lower the activation temperature threshold of TRPV2 to around or even below core body temperature. In the present study, we investigated whether methionine oxidation activates TRPV2 and regulates thermogenesis in the differentiated brown adipocytes. As a result, treatment with Chloramine-T (ChT, a methionine oxidant) activated TRPV2 at temperatures of >30°C in mouse TRPV2-expressing HEK293T cells and in the differentiated brown adipocytes. Moreover, ChT treatment enhanced the expression of genes related to thermogenesis in the differentiated brown adipocytes. These results suggest that methionine oxidation might activate TRPV2 at around body temperature and increase thermogenesis-related gene expression.

The specific contributions of apoptosis, necrosis, autophagy, and cellular senescence pathways in malignant tumor cells (especially p-53 defective or mutated tumors) are unclear. We evaluated the cell viability, apoptosis, autophagic cell death, and cell senescence of T98G cells irradiated with 6 megavoltage (MV) X-rays at 0, 2, 4, and 8 Gy. The cell-death rate obtained by a colony formation assay was used to derive the cell-surviving fraction at each irradiation dose. We evaluated the relative balance between each irradiation dose and the cell-death pathway by a four-dimensional (4D) plot generated by incorporating the cell-death rate, autophagic cell death, and cell senescence. A dose-dependent decrease in the cell-survival rate was observed, with no significant apoptosis induction at any dose. Autophagic cell death and cellular senescence both exhibited dose-dependent increases, with autophagy dominating at 2-4 Gy and senescence more prominent at 8 Gy. The complex relationships among apoptosis, autophagy, and senescence in T98G remain unresolved. Our results highlight the dose-dependent shifts in autophagy and senescence, providing a foundational understanding of cell-death dynamics in radiation-resistant tumors and perhaps paving the way for novel strategies that exploit these pathways to enhance glioblastoma treatments' efficacy.

The health benefits of apple polyphenols are well-documented. Phlorizin (Pz), an apple-derived polyphenol, is a known inhibitor of sodium-glucose cotransporters (SGLTs); however, the effects of other apple constituents on SGLTs remain unclear. In this study, we examined the inhibitory effects of an unripe apple polyphenol extract, ApplePhenon® (AP), and its constituents-including Pz, phloretin, procyanidins B1 and B2, chlorogenic acid, gallic acid, catechin, epicatechin, epigallocatechin gallate, and quercetin-on SGLTs using the Ussing chamber in mouse small intestinal mucosa. AP exhibited noncompetitive inhibition of glucose-induced short-circuit current (Isc), reflecting electrogenic Na+ transport via SGLTs (apparent Ki = 22.23 ± 3.06 μg·mL-1). A similar inhibitory effect was observed in the case of Pz (Ki = 1.88 ± 0.32 μg·mL-1), phloretin (Ki = 18.82 ± 3.52 μg·mL-1), and quercetin (Ki = 220.9 ± 3.52 μg·mL-1) but not in other components. Based on the total Pz content of 6.2%, 86.0% of the inhibitory effect can be attributed to Pz. Although Pz is known to inhibit SGLT1 competitively, AP and its constituents, including Pz, phloretin, and quercetin, showed noncompetitive inhibition in this study. These findings warrant further investigation into the mechanisms underlying SGLT inhibition by AP.

We investigated the roles of muscular ubiquitin-specific protease (USP) 2 in the integrity of liver and muscle in metabolic dysfunction-associated steatotic liver disease (MASLD) using a mouse model. Choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) for 8 weeks caused apparent steatosis and fibrosis in the liver of C57BL/6N mice, whereas serum triglyceride and total cholesterol levels were reduced. Although CDAHFD promoted steatosis, inflammation, and fibrosis in the liver, it failed to affect the tissue mass of the gastrocnemius and soleus muscles. Muscle-specific Usp2 knockout (musUsp2KO) mice and control Usp2fl/fl mice exhibited the CDAHFD-induced changes in blood lipids, liver weight, and liver histology at a comparable level. Similarly, soleus muscle weight, diameter of muscle fibers, and physical activity were indistinguishable between musUsp2KO mice and Usp2fl/fl mice in CDAHFD-induced MASLD condition. Comprehensive RNA sequencing and subsequent RT-qPCR analyses indicated that USP2 deficiency potentiated C1qtnf3 expression in the muscle of MASLD mice. Accordingly, chemical inhibition of USP2 elevated C1qtnf3 mRNA in C2C12 myotubes. Application of recombinant C1QTNF3 stimulated mitochondrial biogenesis in C2C12 cells. Considering that USP2 mitigates mitochondrial oxidative stress, induction of C1QTNF3 might compensate for the depletion of USP2 in skeletal muscle.

Amyloid β (Aβ) (1‒40) is the major form in amyloid plaques, while Aβ (1‒42) is predominant in neuronal plaques. Anti-Aβ antibodies are clinically accepted for Alzheimer's disease treatment to remove Aβ from neuronal plaques; however, increase of intracranial hemorrhagic risk is a major concern. We reported that Aβ (1‒40) inhibits thrombin receptor-activating protein (TRAP)-induced platelet activation in healthy volunteers, and the responsiveness of Aβ (1‒40) to the platelet activation is related to brain atrophy in diabetes mellitus (DM) patients. We investigated the difference between the effects of Aβ (1‒40) and Aβ (1‒42) on the platelet activation in DM participants. Both isoforms suppressed the platelet aggregation, but the effect of Aβ (1‒42) was smaller than Aβ (1‒40). The effect of Aβ (1‒42) on the TRAP-stimulated phosphorylation of p38 mitogen-activated protein kinase and stress-activated protein kinase/c-Jun N-terminal kinase was smaller than Aβ (1‒40). Although the differences were not clarified, the effective ratio of Aβ (1‒40) to Aβ (1‒42) on the PDGF-AB secretion effect was related to the ratio on the aggregation and the phosphorylated-HSP27 secretion. These results suggest that the difference of the effects exists between Aβ (1‒40) and Aβ (1‒42) on the TRAP-stimulated platelet activation individually in the DM patients.

Myocardial ischemia/reperfusion (I/R) injury can lead to substantial cardiac damage and pose a significant threat to human health and survival. Micheliolide (MCL), a sesquiterpene lactone extracted from Michelia (Magnoliaceae), has been reported to exhibit anti-inflammatory, anti-fibrotic and anti-oxidant properties. However, the potential regulatory role of MCL in the progression of myocardial I/R injury remains unclear. In this study, we demonstrated that MCL improved cell viability in H9c2 cells subjected to oxygen-glucose deprivation/reoxygenation (OGD/R). Furthermore, MCL treatment significantly attenuated the inflammatory response induced by OGD/R in H9c2 cells. OGD/R stimulation markedly increased reactive oxygen species (ROS) production, which was effectively reduced by MCL administration. In addition, MCL inhibited the elevated apoptosis observed in OGD/R-treated H9c2 cells. Mechanistically, MCL suppressed the activation of the NF-κB signaling pathway following OGD/R exposure. In summary, this study provides the first evidence that MCL mitigates OGD/R-induced inflammation and oxidative stress in H9c2 cells, suggesting its potential as a therapeutic candidate for the treatment of myocardial I/R injury.

The distribution of transient receptor potential melastatin 7 (TRPM7) was investigated in the trigeminal (TG), petrosal (PG), jugular (JG), and nodose ganglia (NG) in a rat model. Approximately half of the sensory neurons contained TRPM7-immunoreactivity in the TG, PG, JG, and NG. TRPM7-immunoreactive (-IR) sensory neurons had small- to medium-sized cell bodies. Triple immunofluorescence analysis revealed that TRPM7-IR sensory neurons showed mainly transient receptor potential vanilloid 1 (TRPV1) immunoreactivity, whereas less than 10% of TRPM7-IR neurons contained TRPV2-immunoreactivity. In the facial skin, some nerve fibers contained TRPM7-immunoreactivity and formed free nerve endings. Using a retrograde tracing method, 56% of cutaneous TG neurons contained TRPM7-immunoreactivity. Approximately half of TRPM7-IR cutaneous TG neurons contained TRPV1-immunoreactivity. Approximately two-thirds of PG, JG, and NG neurons innervating the external ear canal skin contained TRPM7-immunoreactivity. Most TRPM7-IR cutaneous PG, JG, and NG neurons contained TRPV1-immunoreactivity. However, TRPM7-IR neurons that innervate the tooth pulp and pharynx are relatively rare. This study suggests that the distribution of TRPM7 depends on the size of nerve cells and peripheral innervation. TRPM7 may be involved in pain sensation in the trigeminal, glossopharyngeal, and vagal ganglia sensory neurons.

The gastrointestinal (GI) tract serves as a dynamic chemosensory interface that integrates signals from dietary phytochemicals and microbiota-derived metabolites to regulate host physiology. Beyond digestion and absorption, specialized epithelial and enteroendocrine cells detect luminal compounds via receptors such as taste (sweet, bitter), olfactory, and transient receptor potential (TRP) channels. Phytochemicals - including terpenoids, glycosides, flavonoids, and volatile compounds - activate these receptors to modulate gut hormone secretion, appetite, energy balance, and immune function. Similarly, microbiota-derived metabolites such as short-chain fatty acids, bile acids, and tryptophan derivatives act through G protein-coupled and nuclear receptors to coordinate metabolic, immune, and neuroendocrine processes. Together, these receptor-mediated pathways form a complex communication network linking diet, microbes, and host signaling systems, influencing metabolic health and disease. Future research integrating multi-omics and advanced imaging is expected to clarify these molecular interactions and support the development of precision nutrition strategies targeting gut chemosensory systems for the prevention and treatment of obesity, diabetes, and related disorders.

Trace amine-associated receptor 1 (TAAR1) is highly expressed in rat parotid acinar cells; however, its role in exocrine Ca2+ signaling remains unclear. We herein demonstrated that the selective TAAR1 agonist RO5256390 induced rapid and transient increases in intracellular Ca2+ levels that were completely abolished by the TAAR1 antagonist EPPTB, confirming receptor specificity. Pharmacological dissection revealed dual contributions from extracellular Ca2+ influx via L-type, T-type, and receptor-operated channels and Ca2+ release from intracellular stores mediated by phospholipase C, inositol trisphosphate receptors, and ryanodine receptors. Inhibition of adenylyl cyclase or protein kinase A (PKA) nearly abolished the Ca2+ response, and displacement of PKA from AKAPs using the cell-permeable AKAP-displacing peptide st-Ht31 produced a comparable suppression, underscoring the requirement for microdomain-restricted PKA activity. Exchange protein directly activated by cAMP 2 (Epac 2) and downstream calmodulin-dependent protein kinase II were also indispensable, whereas Epac 1 was dispensable. The present study identified a TAAR1-centered, 3',5'-cyclic adenosine monophosphate microdomain network that co-ordinates Ca2+ entry and release, thereby providing novel targets for the modulation of salivary secretion.

Radiological and nuclear accidents require reliable biomarkers for rapid detection of radiation exposure and dose estimation. While conventional biodosimetry has focused on nuclear DNA damage in lymphocytes, mitochondrial DNA (mtDNA) may provide an additional index because of its high copy number and limited repair capacity. Previous studies using Epstein-Barr virus-transformed lymphocytes and HeLa-FUCCI cells demonstrated radiation-induced changes in mtDNA copy number (mtDNAcn), suggesting compensatory replication as a characteristic response, but in vivo evidence has been limited. In this study, we analyzed peripheral blood from C57BL/6N male mice (8 weeks old) exposed to 0, 0.05, 0.2, 0.5, or 2 Gy of X-rays. MtDNAcn and intact copy ratio, defined as the proportion of undamaged copies, were quantified at 1 day and 1 week post-irradiation. We found that mtDNAcn was significantly increased only in the 2 Gy group at both 1 day and 1 week, whereas the intact copy ratio was significantly decreased in the 0.5 Gy and 2 Gy groups at 1 day but returned to baseline by 1 week. These findings indicate that peripheral blood-derived mtDNA indices are promising biomarkers for radiation biodosimetry, and that intact copy ratio may be particularly useful for detecting low-dose exposure.

Finerenone is a novel non-steroidal mineralocorticoid receptor antagonist and exhibits anti-fibrotic effects against diabetic nephropathy (DN), but its underlying mechanism of action remains unclear. This study aimed to elucidate the effects of Finerenone on high glucose (HG)-induced fibrosis in HK-2 cells and explore the underlying mechanisms. Finerenone was used to treat HG-stimulated HK-2 cells. ELISA and immunofluorescence assays were used to evaluate extracellular matrix (ECM) accumulation and epithelial-to-mesenchymal transition (EMT). Western blotting and molecular docking were performed to investigate the interaction of Finerenone with TGF-β1/Smad signaling. ALK5 knockdown experiments were conducted to confirm Finerenone's target specificity. Finerenone significantly reduced HG-induced fibronectin, collagen III, collagen IV and α-SMA expression. Moreover, Finerenone restored E-cadherin while suppressed N-cadherin and vimentin levels. Notably, it did not alter TGF-β1 production, but inhibited Smad2/3 phosphorylation. Molecular docking showed its competitively binding site to TGF-βR1, knockdown of which abolished Finerenone's anti-fibrotic effects. Finerenone mitigates HG-induced fibrosis in HK-2 cells by targeting TGF-βR1 and inhibiting downstream Smad signaling to modulate ECM accumulation and EMT. These findings provided mechanistic insights into Finerenone's potential as a therapeutic agent for DN.

Cluster of differentiation 36 (CD36) is a cell-surface receptor that exhibits multifunctional properties with diverse ligands. A short segment of CD36, consisting of amino acids 149-168 (CD36149-168), has been shown to constitute a site for recognizing its lipid ligands (e.g., distinct forms of oxidized phospholipids). Recently, lauric acid was found to bind to the CD36 segment. Of interest, the fatty acid and some of its derivatives such as laurylamine and 1-monolaurin are well known to have a cytotoxic activity. This study aimed to assess whether lipids with cytotoxicity, including lauric acid derivatives, could be CD36 ligands by conducting an assay in which a fluorescently-labeled peptide containing human CD36149-168 was utilized. The assay results indicated that laurylamine and 1-monolaurin could bind to CD36149-168. Conversely, no evidence was obtained for non-cytotoxic derivatives (e.g., lauryl mercaptan) interacting with the CD36 segment. Other cytotoxic fatty amines (e.g., myristylamine) and 1-monoglycerides (e.g., 1-monocaprin) were also shown to be recognized by CD36149-168. Identification of these lipids offers an additional category for CD36 ligands, namely, "cytotoxic lipids," which further supports the role of this receptor as a multipurpose player.

The aim of this study is to clarify whether adult pancreatic islet cells would strictly maintain the ability to produce a single pancreatic hormone, or whether they would retain the plasticity to produce two or more pancreatic hormones simultaneously. The ability to produce pancreatic hormones was estimated by the number of hormone gene transcripts, which was evaluated by a spatial transcriptomics analysis using Xenium. The spatial analysis was performed using 477 probes on pancreatic tissues surgically removed from three patients with pancreatic tumors. Eleven pancreatic islets were selected from pancreatic tissue sections, and a detailed analysis was performed on the number of glucagon, insulin, somatostatin and pancreatic polypeptide gene transcripts in the total of 773 pancreatic islet cells. The spatial analysis revealed that a large number of pancreatic islet cells possessed multiple gene transcripts of four pancreatic hormones. Based on these results, as far as gene expression is concerned, it is possible to speculate that adult pancreatic islet cells have the plasticity to produce two or more pancreatic hormones simultaneously. These results suggested that the hormone-producing capabilities of the conventional A, B, D and PP pancreatic islet cells would need to be re-examined.

Sepsis-associated encephalopathy (SAE) is a critical neurological complication of sepsis with limited therapeutic options. This study investigated the neuroprotective potential of remimazolam (REMI) and dexmedetomidine (DEX) in a lipopolysaccharide (LPS)-induced murine model of SAE. We assessed cognitive function via trace fear conditioning, hippocampal CA3 neuronal integrity, neuroinflammatory markers (TNF-α, IL-6, IL-1β), blood-brain barrier (BBB) permeability through hippocampal albumin levels, and lung Netrin-1 expression as an indicator of vagus nerve pathway activity. LPS administration resulted in significant cognitive deficits, CA3 neuronal damage, elevated systemic and central pro-inflammatory cytokines, increased BBB permeability, and reduced lung Netrin-1. Pre-treatment with either REMI or DEX substantially ameliorated these LPS-induced neuropathological changes and cognitive impairments. Notably, the protective effects of both agents on cognitive function were significantly attenuated by the α7 nicotinic acetylcholine receptor (α7nAChR) antagonist, methyllycaconitine. These findings suggest that REMI and DEX exert neuroprotective effects against LPS-induced acute brain injury and cognitive decline, likely mediated, at least in part, through the α7nAChR pathway, highlighting their potential therapeutic relevance in mitigating SAE.

Pro-gastrin-releasing peptide (ProGRP) is the precursor of gastrin-releasing peptide (GRP), a neuropeptide hormone with diverse biological activities. ProGRP is an established tumor marker for small cell lung carcinoma (SCLC). Recently, we reported the usefulness of ProGRP as a tumor marker for Ewing's sarcoma (ES) family tumors (ESFTs). We performed in vitro and in vivo studies on the expression of ProGRP in ES cell lines to assess the reliability of ProGRP as a tumor marker for ESFTs. High levels of GRP mRNA and ProGRP protein were detected in ES cell lines, including A-673, SK-N-MC, SK-NEP-1, and DMS 53 SCLC cell line, but no expression was detected in SU-CCS-1 clear cell sarcoma cell line. ProGRP expression was also detected in in vivo studies using tumor-bearing athymic mice inoculated with ES and SCLC cell lines. In tumor growth experiment using A-673 cells, plasma ProGRP was detected in all mice one week after tumor cell inoculation. Plasma ProGRP levels increased with increasing tumor size, and surgical resection of the tumor on Day 21 eliminated plasma ProGRP completely. These results demonstrate that ProGRP is produced and released by ES cells and strongly indicate the high ability of ProGRP as a diagnostic marker for ESFTs.

The translocator protein (TSPO) is a mitochondrial outer membrane protein that is constitutively expressed in various immune cells, including macrophages, dendritic cells, T cells, and B cells. TSPO has been implicated in mitochondrial function and immune regulation, particularly in macrophages, which are also osteoclast precursors. However, its role in osteoclast differentiation remains unclear. This study examined the effects of the TSPO ligand Ro5-4864 on osteoclast differentiation using murine bone marrow-derived macrophages (BMMs) and the RAW 264.7 macrophage cell line. TSPO was confirmed to be expressed constitutively in BMMs and remained detectable after stimulation with receptor activator of NF-κB ligand (RANKL), consistent with a potential role in osteoclastogenesis. Treatment with Ro5-4864 suppressed RANKL-induced osteoclast differentiation in both BMMs and RAW264.7 cells. This suppression of osteoclast differentiation was accompanied by downregulation of osteoclast-associated genes, including Nfatc1, Acp5, Mmp9, and Ctsk. Metabolic analysis revealed that Ro5-4864 decreased cellular ATP levels without altering lactate production. Ro5-4864 also increased the mitochondrial membrane potential. Although the specific role of TSPO in osteoclastogenesis remains unclear, our findings suggest that Ro5-4864 inhibits osteoclast differentiation via mechanisms related to TSPO and mitochondrial energy metabolism.

During the delayed emesis phase, methotrexate tended to increase 5-hydroxytryptamine (5-HT) levels in midbrain and significantly increased 5-hydroxyindole-acetic acid (5-HIAA), a metabolite of 5-HT, levels in the area postrema and in the cerebral cortex in rats. However, methotrexate did not increase mRNA expression of 5-HT biosynthesis/metabolism enzyme in the area postrema. The increased 5-HIAA levels in the area postrema may result from enhanced intestinal 5-HT synthesis, with 5-HIAA reaching the area postrema via circulation. Additionally, the rise in cortical 5-HIAA levels might reflect stress-related increases in brain tryptophan influx.