Search Results (42)

The Polydimethylsiloxane (PDMS) SlipChip is a microfluidic platform enabling fluid manipulation without pumps or valves, simplifying operation and reducing reagent use. High-viscosity silicone oils (e.g., 5000–10,000 cSt) improve sealing but frequently block microfluidic channels, reducing usability. In contrast, low-viscosity oils (50–100 cSt) reduce blockages but may compromise sealing. This study addresses these challenges by optimizing the viscosity of silicone oil and the curing conditions of PDMS. Low-viscosity silicone oil (50 cSt) was identified as optimal, ensuring smooth slipping and reliable sealing without blockages. Curing conditions were also adjusted to balance adhesion and stiffness as follows: lower temperatures (50–60 °C) enhanced van der Waals adhesion, while higher temperatures (80 °C) increased stiffness. A mixed curing approach (80 °C for the top layer and 60 °C for the bottom layer) further improved performance. Biocompatibility testing using human osteosarcoma cells demonstrated minimal cytotoxicity with 50 cSt oil, supporting cell viability (95%) comparable to traditional multiwell plates. These findings provide practical guidelines for fabricating reliable and biocompatible SlipChips. Full article

Background/Objectives: Postoperative analgesia is important for reducing biologically invasive reactions to surgery. In Japan, postoperative analgesia, including indices of analgesia, has not been adequately addressed. This study aimed to determine the relationship between postoperative pain and postoperative course and the importance of analgesia for early recovery. Methods: Patients who underwent any of seven surgical procedures in gastrointestinal, thoracic, and cardiac surgery were enrolled. The primary endpoint was a median Prince Henry Pain Scale score from postoperative days 1 to 3. Secondary endpoints were the quality of recovery on postoperative day 7 (Quality of Recovery-15 [QoR-15]) and the length of postoperative hospital stay. Results: Median postoperative pain levels among surgeries were 3 on day 1, 2 on days 2 and 3, 1 on day 7, and 1 at discharge. In both univariate and multivariate analyses, the use of postoperative epidural analgesia and intravenous patient-controlled analgesia (IV-PCA) were significant predictors of early postoperative pain. Only early postoperative pain was a significant predictor of QoR-15 score. Regular use of acetaminophen, early postoperative pain, no appetite, and postoperative complications were significant in affecting the length of postoperative hospital stay. In the comparison of early postoperative pain according to whether epidural analgesia and IV-PCA were used, the group that used both methods had the least pain. Conclusions: In Japan, early postoperative pain persists after major surgical procedures and affects postoperative quality of recovery and length of hospital stay. The use of epidural analgesia, IV-PCA, or both appeared to be effective in overcoming early postoperative pain, thereby enhancing early postoperative recovery. Full article

Background and Objectives: We analyzed the changes in obesity, glucose metabolism, and body composition over a 3-year period in Japanese patients with obesity following laparoscopic sleeve gastrectomy (LSG). Methods: Body weight, parameters related to diabetes such as glycated hemoglobin (HbA1c), and electrical impedance analysis were used to assess body composition in forty-eight Japanese patients with obesity before surgery and 6 months, 1 year, 2 years, and 3 years after LSG. Results: At 6 months, 1, 2, and 3 years post-LSG, there were significant reductions in body weight, body mass index, blood pressure, fasting plasma glucose, triglyceride, and HbA1c levels. Six months after LSG, fat mass (FM), muscle mass (MM), and %FM all showed a decrease compared to pre-treatment values (all p < 0.05). FM and %FM remained in a decreased state until 3 years had passed. In contrast, %MM increased at 6 months post-LSG and was maintained up to 3 years post-LSG (all p < 0.05). Furthermore, changes in FM and %FM were associated with changes in body weight and A1C. In contrast, change in %MM exhibited a negative correlation with body weight and A1C following LSG. Finally, multivariate regression analyses demonstrated that alterations in FM were independent factors affecting body weight in patients with obesity 3 years after LSG. Conclusions: We observed improvements in FM, fasting plasma glucose, and HbA1c levels over a 3-year period in Japanese patients after LSG. The reduction in FM and maintenance of %MM after LSG were suggested as possible links between the effects of LSG on obesity and diabetes over 3 years. Full article

In this study, with the aim of adapting an enzyme-linked immunosorbent assay (ELISA) system for point-of-care testing (POCT), we propose an image analysis method for ELISAs using a centrifugal microfluidic device that automatically executes the assay. The developed image analysis method can be used to quantify the color development reaction on a TMB (3,3′,5,5′-tetramethylbenzidine) substrate. In a conventional ELISA, reaction stopping reagents are required at the end of the TMB reaction. In contrast, the developed image analysis method can analyze color in the color-developing reaction without a reaction stopping reagent. This contributes to a reduction in total assay time. The microfluidic devices used in this study could execute reagent control for ELISAs by steady rotation. In the demonstration of the assay and image analysis, a calibration curve for mouse IgG detection was successfully prepared, and it was confirmed that the image analysis method had the same performance as the conventional analysis method. Moreover, the changes in the amount of color over time confirmed that a calibration curve equal to the endpoint analysis was obtained within 2 min from the start of the TMB reaction. As the assay time before the TMB reaction was approximately 7.5 min, the developed ELISA system could detect TMB in just 10 min. In conventional methods using a plate reader, the assay required a time of 90 min for manual handling using microwell plates, and in the case of using automatic microfluidic devices, 30 min were required. The time of 10 min realized by this proposed method is equal to the time required for detection in an immunochromatographic assay with a lateral flow assay; therefore, it is expected that ELISAs can be performed sufficiently to adapt to POCT. Full article

Osteoclasts are bone-resorbing multinucleated giant cells formed by the fusion of monocyte/macrophage lineages. Various small GTPases are involved in the multinucleation and differentiation of osteoclasts. However, the roles of small GTPases regulatory molecules in osteoclast differentiation remain unclear. In the present study, we examined the role of Dennd2c, a putative guanine nucleotide exchange factor for Rab GTPases, in osteoclast differentiation. Knockdown of Dennd2c promoted osteoclast differentiation, resorption, and expression of osteoclast markers. Morphologically, Dennd2c knockdown induced the formation of larger osteoclasts with several protrusions. In contrast, overexpression of Dennd2c inhibited the multinucleation and differentiation of osteoclasts, bone resorption, and the expression of osteoclast markers. Dennd2c-overexpressing macrophages exhibited spindle-shaped mononuclear cells and long thin protrusions. Treatment of Dennd2c-overexpressing cells with the Cdc42 inhibitor ML-141 or the Rac1 inhibitor 6-thio-GTP prevented protrusion formation. Moreover, treatment of Dennd2c-overexpressing cells with the actin polymerization inhibitor latrunculin B restored multinucleated and TRAP-positive osteoclast formation. These results indicate that Dennd2c negatively regulates osteoclast differentiation and multinucleation by modulating protrusion formation in macrophages. Full article

It is believed that short-chain fatty acids (SCFAs)—the bacterial metabolites produced by the digestion of dietary fiber—potentially contribute to the prevention of colitis. However, this beneficial effect has not been conclusively proven. We thus attempted to verify this beneficial effect by examining whether colitis can be caused or worsened by the deficiency in dietary fiber in mice. We found that dextran sodium sulfate (DSS)-induced colitis was accelerated under a fiber-deficient condition, but the fiber deficiency itself did not provoke colitis. Moreover, episodes of diarrhea and epithelium damage in the large intestine were found upon analysis 24 h after the intervention. Unfortunately, these symptoms and tissue damage could not be ameliorated by administering SCFAs. On the other hand, a fiber-deficient condition increased the population of Desulfovibrio spp. and decreased the population of Lactobaccillus spp. regardless of the presence or absence of DSS upon analysis 24 h after the intervention. These results suggest that a deficiency in dietary fiber makes the intestinal environment irritable to colitis-inducing stimuli within the short term. This change does not appear to be related to the presence of SCFAs, but to the alteration of microbiota. Hence, a regular intake of dietary fiber is strongly recommended to avoid colitis and preserve intestinal health. Full article

(1) Background: Exosomal PD-L1 has garnered attention owing to its role in instigating systemic immune suppression. The objective of this study is to elucidate whether bone and soft tissue sarcoma cells possess the capacity to secrete functionally active exosomal PD-L1 and whether radiotherapy (RT) induces the exosomal PD-L1 release. (2) Methods: Human osteosarcoma cell line 143B and human fibrosarcoma cell line HT1080 were utilized. Exosomes were isolated from the culture medium and blood via ultracentrifugation. The expression of PD-L1 on both tumor cells and exosomes was evaluated. The inhibitory effect on PBMC was employed to assess the activity of exosomal PD-L1. Post radiotherapy, changes in PD-L1 expression were compared. (3) Results: Exosomal PD-L1 was detected in the culture medium of tumor cells but was absent in the culture medium of PD-L1 knockout cells. Exosomal PD-L1 exhibited an inhibitory effect on PBMC activation. In tumor-bearing mice, human-derived exosomal PD-L1 was detected in the bloodstream. Following radiotherapy, tumor cells upregulated PD-L1, and human-derived exosomal PD-L1 were detected in the bloodstream. (4) Conclusions: Exosomal PD-L1 emanates from bone and soft tissue sarcoma cells and is disseminated into the circulatory system. The levels of PD-L1 in tumor cells and the release of exosomal PD-L1 were augmented after irradiation with RT. Full article

This study evaluated the potential of gelatin methacryloyl (GelMA) for single-cell screening compared to polyethylene glycol diacrylate (PEGDA). GelMA photopolymerized at 1000–2000 mJ/cm² produced consistent patterns and supported HeLa cell viability. GelMA (5%w/v) facilitated better cell collection within 2 days due to its shape retention. GelMA demonstrated biocompatibility with HeLa cells exhibiting exponential proliferation and biodegradation over 5 days. The average cell displacement over 2 days was 16 µm. Two targeted cell recovery strategies using trypsin were developed: one for adherent cells encapsulated at 800 mJ/cm², and another for floating cells encapsulated at 800 mJ/cm², enabling the selective removal of unwanted cells. These findings suggest GelMA as a promising biomaterial for single-cell screening applications, offering advantages over PEGDA in cell encapsulation and targeted recovery. Full article

Background: Chronic obstructive pulmonary disease (COPD) typically causes airflow blockage and breathing difficulties, which may result in the abnormal morphology and motion of the lungs or diaphragm. Purpose: This study aims to quantitatively evaluate respiratory diaphragm motion using a thoracic sagittal magnetic resonance imaging (MRI) series, including motion asynchronization and limitations. Method: First, the diaphragm profile is extracted using a deep-learning-based field segmentation approach. Next, by measuring the motion waveforms of each position in the extracted diaphragm profile, obvious differences in the independent respiration cycles, such as the period and peak amplitude, are verified. Finally, focusing on multiple breathing cycles, the similarity and amplitude of the motion waveforms are evaluated using the normalized correlation coefficient (NCC) and absolute amplitude. Results and Contributions: Compared with normal subjects, patients with severe COPD tend to have lower NCC and absolute amplitude values, suggesting motion asynchronization and limitation of their diaphragms. Our proposed diaphragmatic motion evaluation method may assist in the diagnosis and therapeutic planning of COPD. Full article

A new generation of nanoscale photosensitizer agents has improved photothermal capabilities, which has increased the impact of photothermal treatments (PTTs) in cancer therapy. Gold nanostars (GNS) are promising for more efficient and less invasive PTTs than gold nanoparticles. However, the combination of GNS and visible pulsed lasers remains unexplored. This article reports the use of a 532 nm nanosecond pulse laser and polyvinylpyrrolidone (PVP)-capped GNS to kill cancer cells with location-specific exposure. Biocompatible GNS were synthesized via a simple method and were characterized under FESEM, UV–visible spectroscopy, XRD analysis, and particle size analysis. GNS were incubated over a layer of cancer cells that were grown in a glass Petri dish. A nanosecond pulsed laser was irradiated on the cell layer, and cell death was verified via propidium iodide (PI) staining. We assessed the effectiveness of single-pulse spot irradiation and multiple-pulse laser scanning irradiation in inducing cell death. Since the site of cell killing can be accurately chosen with a nanosecond pulse laser, this technique will help minimize damage to the cells around the target cells. Full article

Aim: Primary malignant bone tumor osteosarcoma can metastasize to the lung. Diminishing lung metastasis would positively affect the prognosis of patients. Our previous studies demonstrated that highly metastatic osteosarcoma cell lines are significantly softer than low-metastasis cell lines. We therefore hypothesized that increasing cell stiffness would suppress metastasis by reducing cell motility. In this study, we tested whether carbenoxolone (CBX) increases the stiffness of LM8 osteosarcoma cells and prevents lung metastasis in vivo. Methods: We evaluated the actin cytoskeletal structure and polymerization of CBX-treated LM8 cells using actin staining. Cell stiffness was measured using atomic force microscopy. Metastasis-related cell functions were analyzed using cell proliferation, wound healing, invasion, and cell adhesion assays. Furthermore, lung metastasis was examined in LM8-bearing mice administered with CBX. Results: Treatment with CBX significantly increased actin staining intensity and stiffness of LM8 cells compared with vehicle-treated LM8 cells (p < 0.01). In Young’s modulus images, compared with the control group, rigid fibrillate structures were observed in the CBX treatment group. CBX suppressed cell migration, invasion, and adhesion but not cell proliferation. The number of LM8 lung metastases were significantly reduced in the CBX administration group compared with the control group (p < 0.01). Conclusion: In this study, we demonstrated that CBX increases tumor cell stiffness and significantly reduces lung metastasis. Our study is the first to provide evidence that reducing cell motility by increasing cell stiffness might be effective as a novel anti-metastasis approach in vivo. Full article

The combination of the mammalian target of rapamycin and proteasome inhibitors is a new treatment strategy for various tumors. Herein, we investigated the synergistic effect of everolimus and bortezomib on tumor growth and metastasis in bone and soft tissue sarcomas. The antitumor effects of everolimus and bortezomib were assessed in a human fibrosarcoma (FS) cell line (HT1080) and mouse osteosarcoma (OS) cell line (LM8) by MTS assays and Western blotting. The effects of everolimus and bortezomib on HT1080 and LM8 tumor growth in xenograft mouse models were evaluated using tumor volume and the number of metastatic nodes of the resected lungs. Immunohistochemistry was used to evaluate cleaved PARP expression. The combination therapy decreased FS and OS cell proliferation compared with either drug alone. This combination induced more intense p-p38, p-JNK, and p-ERK and activated apoptosis signals, such as caspase-3, compared with single-agent treatment. The combination treatment reduced p-AKT and MYC expression, decreased FS and OS tumor volumes, and suppressed lung metastases of OS. The combination therapy inhibited tumor growth in FS and OS and metastatic progression of OS via the JNK/p38/ERK MAPK and AKT pathways. These results could aid in the development of new therapeutic strategies for sarcomas. Full article

The association between oral frailty (OFr) and body action has been investigated, but its association with systemic function remains unclear. Therefore, this cross-sectional study examined the association between OFr with decreased bone mineral density (BMD) and renal function in residents of Shika town, Ishikawa Prefecture, Japan aged ≥40 years. This study included 400 inhabitants. The OFr total score was assessed using three oral domains in the Kihon Checklist (a self-reported comprehensive health checklist), the number of teeth, and brushing frequency per day. Measurements were the estimated glomerular filtration rate (eGFR) and the osteo-sono assessment index (OSI). Using a two-way analysis of covariance (p = 0.002), significantly lower OSI was indicated in the eGFR < 60 and OFr group than in the eGFR of < 60 and non-OFr group after adjusting for age, body mass index, and drinking and smoking status as confounding factors. Multiple logistic regression analysis confirmed this relationship (p = 0.006). Therefore, lower BMD seems to be associated with lower renal function only when accompanied by OFr. Further longitudinal studies are needed to confirm these results. Full article

Rab44 was recently identified as an atypical Rab GTPase that possesses EF-hand and coiled-coil domains at the N-terminus, and a Rab-GTPase domain at the C-terminus. Rab44 is highly expressed in immune-related cells such as mast cells, macrophages, osteoclasts, and granulocyte-lineage cells in the bone marrow. Therefore, it is speculated that Rab44 is involved in the inflammation and differentiation of immune cells. However, little is known about the role of Rab44 in inflammation. In this study, we showed that Rab44 was upregulated during the early phase of differentiation of M1- and M2-type macrophages. Rab44-deficient mice exhibited impaired tumor necrosis factor alpha and interleukin-10 production after lipopolysaccharide (LPS) stimulation. The number of granulocytes in Rab44-deficient mice was lower, but the lymphocyte count in Rab44-deficient mice was significantly higher than that in wild-type mice after LPS stimulation. Moreover, Rab44-deficient macrophages showed impaired nickel-induced toxicity, and Rab44-deficient mice showed impaired nickel-induced hypersensitivity. Upon nickel hypersensitivity induction, Rab44-deficient mice showed different frequencies of immune cells in the blood and ears. Thus, it is likely that Rab44 is implicated in immune cell differentiation and inflammation, and Rab44 deficiency induces impaired immune responses to nickel allergies. Full article

Background and Objectives: This study aimed to investigate the changes in obesity severity, glucose metabolism, and body composition in patients with obesity and type 2 diabetes mellitus treated with glucagon-like peptide 1 receptor agonist (GLP1-RA) semaglutide. Materials and Methods: Body weight (BW), metabolic parameters, and body composition were examined before and 3 months after semaglutide administration. The mass of body fat (FM), fat weight percentage (%FM), mass of skeletal muscle (MM), skeletal MM percentage (%MM), and limb muscles were measured using the bioelectrical impedance method. Results: Semaglutide dramatically reduced the weight, the body mass index (BMI), and the levels of the glucose metabolic markers, including fasting blood glucose and hemoglobin A1c, and accelerated the loss of excess BW. FM, MM, and %FM after semaglutide treatment also decreased. Conversely, semaglutide had no effect on the %MM after 3 months. In limb muscle analyses, right upper and lower leg muscle percentages, left upper and lower leg muscles, and the ratios of the lower/upper muscles were maintained by semaglutide treatment. Conclusions: These results suggest that the GLP1-RA semaglutide effectively reduces body adiposity while maintaining the MM in obese type 2 diabetic patients. Full article