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17 pages, 2956 KiB  
Article
Impact of Photobiomodulation on the Pro-Osteogenic Activity of Dental Pulp Mesenchymal Stem/Stromal Cells
by Marcella Rodrigues Ueda Fernandes, Gabriella Teti, Valentina Gatta, Aurora Longhin, Ana Cecilia Corrêa Aranha and Mirella Falconi
Int. J. Mol. Sci. 2025, 26(17), 8174; https://doi.org/10.3390/ijms26178174 - 22 Aug 2025
Abstract
Photobiomodulation (PBM) consists of applying low-level laser light to biological tissues, leading to modulation of cellular functions. PBM has recently gained much attention in the field of regenerative dentistry thanks to its powerful effect on tissue repair and regeneration. Dental pulp mesenchymal stem/stromal [...] Read more.
Photobiomodulation (PBM) consists of applying low-level laser light to biological tissues, leading to modulation of cellular functions. PBM has recently gained much attention in the field of regenerative dentistry thanks to its powerful effect on tissue repair and regeneration. Dental pulp mesenchymal stem/stromal cells (DP-MSCs) represent the ideal targets in regenerative dentistry due to their ability to stimulate the regeneration of mineralized and soft tissues and the paracrine factors that they produce. Although there have been several studies evaluating the influence of PBM on DP-MSCs’ regenerative capacity, the results are conflicting, and there are few studies on the influence of PBM on the paracrine factors released by DP-MSCs. Therefore, the aim of this study was to investigate the effect of PBM, using different energy doses of laser irradiation, on the osteogenic capacity of DP-MSCs, focusing on changes in gene expression, mineralizing ability, and release of pro-osteogenic factors. DP-MSCs were irradiated in vitro and differentiated into an osteogenic phenotype. A cell viability assay, alizarin red staining, and TEM analysis were carried out to evaluate the effect of PBM on cell activity, morphology, and mineralization ability. The expression of the main osteogenesis-related markers Runx2, Col1A1, ALP, and BMP was measured to evaluate the influence of PBM on the ability of DP-MSCs to differentiate toward an osteogenic phenotype. The release of IL-6 and IL-8, which are mainly involved in bone remodeling processes, was investigated in the cell medium following PBM irradiation. The results showed a high level of cell viability, suggesting a lack of phototoxicity under the tested conditions. Furthermore, PBM had a significant effect on mineral deposition, IL-6 and IL-8 release, and expression of osteogenic markers. TEM analysis showed intracellular modifications linked mainly to mitochondria, the endoplasmic reticulum, and autophagic vesicles after PBM treatment. These findings demonstrated that the impact of PBM on the osteogenic potential of DP-MSCs is energy dose-dependent, supporting its potential as an effective strategy in regenerative dentistry, particularly for enhancing bone remodeling. Full article
(This article belongs to the Special Issue Application of Biotechnology to Dental Treatment)
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15 pages, 3387 KiB  
Article
Sodium Cuminate Inhibits the Mycelial Growth of Penicillium digitatum by Inducing Oxidative Stress and Damaging the Cell Membrane
by Mingchen Yang, Yonghua Zhang, Xiaoli Tan, Lu Li, Qiuli OuYang and Nengguo Tao
J. Fungi 2025, 11(9), 612; https://doi.org/10.3390/jof11090612 - 22 Aug 2025
Abstract
Green mold formed by Penicillium digitatum is a major disease that limits the yield and overall value of postharvest citrus fruits. The antifungal activity of sodium cuminate (SC) against P. digitatum and the corresponding mechanism were explored in this research. The minimal inhibitory [...] Read more.
Green mold formed by Penicillium digitatum is a major disease that limits the yield and overall value of postharvest citrus fruits. The antifungal activity of sodium cuminate (SC) against P. digitatum and the corresponding mechanism were explored in this research. The minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of SC against P. digitatum were 0.4 and 0.8 g L−1, respectively. SC (8 × MFC) reduced the incidence of disease in Ponkan fruits without compromising their quality. The results of CFW staining and extracellular alkaline phosphatase assays revealed that 1/2MIC SC for 30 min had no impact on the cell wall integrity of P. digitatum. In contrast, 1/2MIC SC apparently destroyed cell membrane integrity, as shown by the increase in the content of reactive oxygen species (ROS), malondialdehyde, and H2O2. The addition of exogenous cysteine (Cys) or diphenyleneiodonium chloride (DPI) significantly mitigated the cytotoxic effects of SC. At the same time, mitochondrial membrane potential was significantly decreased by 1/2MIC SC, and the addition of exogenous Cys or DPI restored it to normal levels. In summary, the antifungal capacity of SC might be attributable to membrane damage in P. digitatum caused by oxidative stress. Full article
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18 pages, 3699 KiB  
Article
Magnolia figo Extract Induces Enamel Shade Recovery and Inhibits Porphyromonas gingivalis Biofilm Formation: An In Vitro, Dual-Action Natural Therapeutic Approach
by Chun-Sheng Kuo, Cheng-Wen Lin, Yuan-Man Hsu, Jen-Chieh Tsai and Dan-Jae Lin
Int. J. Mol. Sci. 2025, 26(17), 8157; https://doi.org/10.3390/ijms26178157 - 22 Aug 2025
Abstract
Dental enamel discoloration, extrinsic staining, and periodontal biofilms remain persistent challenges in oral health. This study explores the in vitro, dual-functional potential of Magnolia figo flower extract (FMO), a sesquiterpene-rich botanical active phytochemical ingredient (API), for aesthetic and antimicrobial oral applications. FTIR identified [...] Read more.
Dental enamel discoloration, extrinsic staining, and periodontal biofilms remain persistent challenges in oral health. This study explores the in vitro, dual-functional potential of Magnolia figo flower extract (FMO), a sesquiterpene-rich botanical active phytochemical ingredient (API), for aesthetic and antimicrobial oral applications. FTIR identified characteristic terpenoid and long-chain fatty acid functional groups, including β-elemene, γ-elemene, and caryophyllene oxide. Whitening efficacy on coffee-stained bovine enamel was quantified using CIELAB colorimetry. The 0.5% FMO treatment achieved ΔE* = 8.49, which was within the clinical perceptibility threshold and the optimal biocompatibility balance. SEM confirmed no demineralization on the enamel surface after immersion in 3.0% FMO for 12 h. Antimicrobial assays demonstrated inhibition of Porphyromonas gingivalis, with MIC and MBC values of 0.25% and 0.5%, respectively. Biofilm formation was reduced by over 50% at a 0.148% concentration. Cytocompatibility assays using HGF-1 cells with various concentrations of FMO showed reduced cell viability at higher concentrations. When exposed for 5 min (simulating daily oral care) or 2 h, 0.5% FMO exhibited greater biocompatibility with L929 cells compared to toothpaste and peroxide-based agents. These findings suggest that FMO may serve as a natural candidate for dual-function oral care; however, further in vivo and clinical investigations are needed to validate its potential use within oral care treatments. Full article
(This article belongs to the Special Issue Natural Compounds in Human Health and Disease)
18 pages, 2144 KiB  
Article
A Semi-Automated and Unbiased Microglia Morphology Analysis Following Mild Traumatic Brain Injury in Rats
by Luke Sumberg, Rina Berman, Antoni Pazgier, Joaquin Torres, Jennifer Qiu, Bodhi Tran, Shannen Greene, Rose Atwood, Martin Boese and Kwang Choi
Int. J. Mol. Sci. 2025, 26(17), 8149; https://doi.org/10.3390/ijms26178149 - 22 Aug 2025
Abstract
Mild traumatic brain injury (mTBI) affects over 40 million people every year. One of its features includes the activation of microglia, the resident immune cells of the brain. Microglia assume different morphological states depending on their level of activation, such as surveilling ramified [...] Read more.
Mild traumatic brain injury (mTBI) affects over 40 million people every year. One of its features includes the activation of microglia, the resident immune cells of the brain. Microglia assume different morphological states depending on their level of activation, such as surveilling ramified and activated hypertrophic, ameboid, and rod-like microglia. These states can be distinguished by multiple features, including the shape, span, and branching of microglia. Male Sprague–Dawley rats sustained mTBI using the Closed-Head Impact Model of Engineered Rotational Acceleration (CHIMERA) (3 times, 1.5 J per impact) or sham treatment. Four days after the injury, brains were collected and stained for microglia using the ionized calcium-binding adapter molecule-1 (Iba-1) antibody. Cortical injury sites were identified in a subset of CHIMERA animals. Using the MicrogliaMorphology ImageJ plugin and the MicrogliaMorphologyR package, 27 morphological features were quantified from individual microglia, and k-means clustering was used to classify microglia as ramified, rod-like, ameboid, and hypertrophic states. The CHIMERA injury altered microglia morphology features, which contributed to increased hypertrophic (activated) and decreased ramified (inactive) microglia compared to the sham controls. Combined with the clinically relevant mTBI paradigm and semi-automated/unbiased approach, the current findings may contribute to microglia morphology classification. Full article
15 pages, 2576 KiB  
Article
Dextromethorphan Enhances Apoptosis and Suppresses EMT in PANC-1 Pancreatic Cancer Cells: Synergistic Effects with Gemcitabine
by Gulsah Medet and Ahmet Inal
Int. J. Mol. Sci. 2025, 26(17), 8151; https://doi.org/10.3390/ijms26178151 - 22 Aug 2025
Abstract
This study aimed to evaluate the effects of dextromethorphan (DX), alone and in combination with gemcitabine (GEM), on cell viability, apoptosis, and epithelial–mesenchymal transition (EMT) markers in PANC-1 human pancreatic cancer cells. PANC-1 human pancreatic cancer cells were cultured and treated with varying [...] Read more.
This study aimed to evaluate the effects of dextromethorphan (DX), alone and in combination with gemcitabine (GEM), on cell viability, apoptosis, and epithelial–mesenchymal transition (EMT) markers in PANC-1 human pancreatic cancer cells. PANC-1 human pancreatic cancer cells were cultured and treated with varying concentrations of dextromethorphan (DX), gemcitabine (GEM), and 5-fluorouracil (5-FU), both as monotherapies and in combination. Cytotoxic effects were assessed using the MTT assay, and IC50 values were calculated at 24, 48, and 72 h. Apoptotic responses were evaluated using Annexin V-FITC/PI staining followed by flow cytometry. Protein expression levels of Bax, Bcl-2, and Vimentin were determined via immunocytochemistry, while EMT markers (E-cadherin, N-cadherin, Vimentin) were analyzed using flow cytometry. Relative mRNA expression of apoptotic and EMT-related genes was quantified by qRT-PCR. DX exhibited time- and dose-dependent cytotoxicity in PANC-1 cells, with IC50 values of 280.4 µM at 24 h, 163.2 µM at 48 h, and 105.6 µM at 72 h. For GEM, the 72 h IC50 was 57.53 µM. The combination of DX 50 µM + GEM 12.5 µM resulted in significantly lower cell viability (24.93 ± 3.12%) compared to GEM 25 µM (35.33 ± 5.22%) and DX 100 µM (51.40 ± 3.10%) (p < 0.001). Flow cytometry revealed significant increases in early (21.83 ± 1.32%) and late apoptotic cells (32.20 ± 0.84%) in the combination group, with a corresponding reduction in viable cells compared to control (24.93 ± 3.12% vs. 89.53 ± 0.97%, p < 0.001). Immunocytochemical analysis showed increased Bax-positive cell count (62.0 cells/unit area), and decreased Bcl-2 (19.0) and Vimentin (28.0) levels in the combination group compared to control (Bax: 15.0, Bcl-2: 60.0, Vimentin: 70.0) (p < 0.001). Flow cytometry for EMT markers demonstrated increased E-cadherin (83.84 ± 0.65%) and decreased Vimentin (71.04 ± 1.17%) and N-cadherin (30.47 ± 0.72%) expression in the DX + GEM group compared to EMT control (E-cadherin: 68.97 ± 1.43%, Vimentin: 91.00 ± 0.75%, N-cadherin: 62.47 ± 1.13%) (p < 0.001). qRT-PCR supported these findings with increased Bax (2.1-fold), E-cadherin (2.0-fold), and reduced Bcl-2 (0.3-fold) and XIAP (0.6-fold) in the combination group (p < 0.05). Dextromethorphan, particularly in combination with gemcitabine, appears to enhance apoptosis and suppress EMT-associated marker expression in PANC-1 cells, supporting its potential as an adjuvant agent in pancreatic cancer therapy. Full article
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15 pages, 1516 KiB  
Article
Association of Enterotoxigenic Bacteroides fragilis with Immune Modulation in Colorectal Cancer Liver Metastasis
by Rumiko Saito, Yasuyuki Shigematsu, Mahmut Amori, Gulanbar Amori, Manabu Takamatsu, Kenji Nishida, Hiroaki Kanda, Yu Takahashi, Yuji Miura, Kengo Takeuchi, Shunji Takahashi and Kentaro Inamura
Cancers 2025, 17(17), 2733; https://doi.org/10.3390/cancers17172733 - 22 Aug 2025
Abstract
Background: Enterotoxigenic Bacteroides fragilis (ETBF) carries the bft toxin gene, which influences the host immune response and inflammatory pathways and promotes colorectal cancer (CRC). This study investigated the potential role of ETBF in CRC liver metastasis. Methods: We reviewed the records [...] Read more.
Background: Enterotoxigenic Bacteroides fragilis (ETBF) carries the bft toxin gene, which influences the host immune response and inflammatory pathways and promotes colorectal cancer (CRC). This study investigated the potential role of ETBF in CRC liver metastasis. Methods: We reviewed the records of 226 consecutive patients who underwent curative-intent (R0) resection of CRC liver metastases. ETBF DNA in fresh-frozen metastasis specimens was quantified using droplet digital PCR (ddPCR). Patients were grouped into very-low (≤80%; N = 178), low (80–90%; N = 24), and high (>90%; N = 24) ETBF-DNA groups. Three tissue cores per specimen were stained for CD8, CD4, CD20, FOXP3, CD68, and CD163, and immune-cell densities were measured digitally (cells/mm2). Results: ETBF DNA was detected in 219 of 226 lesions (96.9%). The densities of cytotoxic CD8+ T-cells, effector CD4+ T-cells, CD20+ B-cells, and CD163+ macrophages did not differ significantly by ETBF-DNA group (Ptrend all > 0.12). FOXP3+ regulatory T-cells (Tregs) decreased (Ptrend = 0.010), and CD68+ macrophages increased (Ptrend = 0.020) as ETBF-DNA levels increased. ETBF-DNA levels in CRC liver metastases were not associated with disease-free survival or overall survival or serum C-reactive protein levels. Conclusions: ETBF was present in almost all CRC liver metastases. Higher ETBF levels were associated with a tumor-immune microenvironment enriched in CD68+ macrophages and deficient in FOXP3+ Tregs, suggesting that ETBF facilitates immune evasion without loss of effector lymphocytes. Although ETBF-DNA levels did not predict survival in this single-center cohort, the potential role of ETBF in immune remodeling and as a candidate biomarker and therapeutic target in metastatic CRC warrants further study. Full article
(This article belongs to the Special Issue Colorectal Cancer Liver Metastases)
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18 pages, 6929 KiB  
Article
4-Propylphenol Alters Membrane Integrity in Fungi Isolated from Walnut Anthracnose and Brown Spot
by Xiaoli Yu, Shuhan Yang, Panhong Su, Haiyao Bi, Yaxuan Li, Xingxing Peng, Xiaohui Sun and Qunqing Wang
J. Fungi 2025, 11(9), 610; https://doi.org/10.3390/jof11090610 - 22 Aug 2025
Abstract
Walnut anthracnose (Colletotrichum gloeosporioides and C. siamense) and brown spot (Alternaria alternata) cause severe yield losses globally. Conventional fungicides face the challenges of pathogen resistance and environmental toxicity. This study evaluates 4-propylphenol, a plant-derived phenolic compound, as an eco-friendly [...] Read more.
Walnut anthracnose (Colletotrichum gloeosporioides and C. siamense) and brown spot (Alternaria alternata) cause severe yield losses globally. Conventional fungicides face the challenges of pathogen resistance and environmental toxicity. This study evaluates 4-propylphenol, a plant-derived phenolic compound, as an eco-friendly alternative against key fungal pathogens of walnut. In vitro assays determined EC50 values against target pathogens (29.11–31.89 mg·L−1) via mycelial growth inhibition and conidial germination suppression (EC50 = 55.04–71.85 mg·L−1). Mechanistic analyses confirmed membrane disruption through propidium iodide staining (9.5-to-14.0-fold fluorescence intensity increase), DNA leakage (77.82–85.15% at 250 mg·L−1), and protein efflux (58.10–66.49%). In field trials, we implemented a phenology-driven strategy: 100 mg·L−1 ground/canopy spray at flowering to reduce primary inoculum, followed by 400 mg·L−1 canopy application at fruiting. This protocol achieved 86.67% control efficacy against disease complexes with negligible phytotoxicity (SPAD variation < 5%). 4-propylphenol provides a sustainable solution through membrane-targeting action, effectively overcoming fungicide resistance in woody crops. Full article
(This article belongs to the Special Issue Plant Pathogens and Mycotoxins)
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13 pages, 2535 KiB  
Article
Effects of Platelet-Rich Fibrin Treated with No-Ozone Cold Plasma on the Alkaline Phosphatase in Rat Bone Marrow Cells: An In Vitro Study
by Byul Bo Ra Choi and Gyoo Cheon Kim
Appl. Sci. 2025, 15(17), 9229; https://doi.org/10.3390/app15179229 - 22 Aug 2025
Abstract
Background/Objectives: Herein, we investigated the effect of platelet-rich fibrin (PRF) treatment combined with no-ozone cold plasma (NCP) on growth factor levels, rat bone-marrow stem cell (rBMSC) proliferation, and alkaline phosphatase (ALP) activity in the early stage of differentiation into osteoblasts. Methods: [...] Read more.
Background/Objectives: Herein, we investigated the effect of platelet-rich fibrin (PRF) treatment combined with no-ozone cold plasma (NCP) on growth factor levels, rat bone-marrow stem cell (rBMSC) proliferation, and alkaline phosphatase (ALP) activity in the early stage of differentiation into osteoblasts. Methods: The PRF used in the experiment was prepared by collecting blood from the jugular vein of rats, followed by centrifugation. The obtained PRF was treated with NCP, and the cell culture media were conditioned with the PRF extracts alone or with NCP-treated PRF extracts. Three different experimental groups were defined: no treatment (NT); cell culture media extracted from PRF (PRF); and cell culture media extracted from PRF treated with NCP (PRF + NCP). Enzyme-linked immunosorbent assays were performed to determine the levels of transforming growth factor (TGF)-β and platelet-derived growth factor (PDGF) AB. Water-soluble Tetrazolium-1 assay was performed to measure cell proliferation in rBMSCs. To analyze cell differentiation into osteoblasts, ALP staining and real-time PCR were performed. Results: Growth factor levels increased in response to treatment (TGF-β: p < 0.001, PDGF AB: p < 0.05), and the cell proliferation rate increased with treatment (145.29% and 150.05% for PRF and the PRF + NCP groups, respectively, relative to the NT group, p < 0.001). Evaluation of the ALP staining intensity and mRNA expression levels showed that the ALP activity was highest in the PRF + NCP group (p < 0.001). Conclusions: Our results confirmed that NCP treatment enhanced the release of several different growth factors contained in PRF to the culture media and that treatment with PRF and NCP increased the proliferation of rBMSCs and their differentiation into osteoblasts. Full article
(This article belongs to the Special Issue Oral Diseases and Clinical Dentistry)
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11 pages, 973 KiB  
Article
Reversible Platelet Aggregation Induced by Low-Temperature Storage in Heparinized Whole Blood Samples
by Yuriko Hayashi, Manato Miyazaki, Ryusuke Kimura, Ririka Arai, Miu Takada, Ayuko Takahashi and Hirokazu Kimura
Hematol. Rep. 2025, 17(5), 42; https://doi.org/10.3390/hematolrep17050042 - 22 Aug 2025
Abstract
Background/Objectives: Platelet counts can be affected by storage conditions, potentially leading to pseudothrombocytopenia. The present study aimed to investigate temperature-dependent changes in platelet counts and morphology in whole blood samples anticoagulated with heparin or EDTA. We also examined the molecular mechanism of [...] Read more.
Background/Objectives: Platelet counts can be affected by storage conditions, potentially leading to pseudothrombocytopenia. The present study aimed to investigate temperature-dependent changes in platelet counts and morphology in whole blood samples anticoagulated with heparin or EDTA. We also examined the molecular mechanism of cold-induced aggregation via integrin GPIIb/IIIa–fibrinogen interaction using established bioinformatics technologies (docking simulation). Methods: Peripheral blood was collected from healthy volunteers (n = 6) and treated with either heparin or EDTA. The samples were stored at 4 °C, room temperature, or incubated at 37 °C. Platelet counts were measured using an automated hematology analyzer. The morphology of various blood cells in smears was assessed using the May-Grünwald Giemsa staining method. Docking simulations using an available software (HADDOCK 2.4) were performed to evaluate integrin–fibrinogen binding at different temperatures. Results: In automated blood cell counting, platelet counts in heparinized blood were significantly decreased under low-temperature conditions (4 °C), but this decrease was restored to levels comparable to those at room temperature upon warming to 37 °C (p < 0.05). No significant changes were observed in EDTA-treated samples. Microscopical findings showed platelet aggregation only in heparinized samples at 4 °C, with normal morphology restored upon warming (37 °C). Docking simulations estimated stronger integrin GPIIb/IIIa–fibrinogen binding at 4 °C than at 37 °C (p = 0.0286), suggesting temperature-dependent enhancement of molecular interactions. Conclusions: These findings indicate that heparin can induce reversible platelet aggregation at low temperatures in whole blood samples, leading to pseudothrombocytopenia. This phenomenon may be mediated by increased integrin GPIIb/IIIa–fibrinogen binding. Full article
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16 pages, 2970 KiB  
Review
Safety and Efficacy of Diquafosol Compared to Artificial Tears for the Treatment of Dry Eye: A Systematic Review and Meta-Analysis
by José Gerardo Serrano-Robles, Ana Karen Pérez-Vázquez, Guillermo Raul Vera-Duarte, Alejandro Navas, Arturo Ramirez-Miranda, Enrique O. Graue-Hernandez and Nicolás Kahuam-López
Int. J. Mol. Sci. 2025, 26(17), 8113; https://doi.org/10.3390/ijms26178113 - 22 Aug 2025
Abstract
Dry eye disease (DED) is a prevalent and disabling condition. Artificial tears are commonly used but often inadequate for moderate-to-severe cases. Secretagogues such as pilocarpine, cevimeline, and diquafosol offer potential alternatives, though their comparative effectiveness remains unclear. To evaluate the safety and efficacy [...] Read more.
Dry eye disease (DED) is a prevalent and disabling condition. Artificial tears are commonly used but often inadequate for moderate-to-severe cases. Secretagogues such as pilocarpine, cevimeline, and diquafosol offer potential alternatives, though their comparative effectiveness remains unclear. To evaluate the safety and efficacy of these secretagogues versus artificial tears in adults with DED, we searched CENTRAL, PubMed, Scopus, LILACS, ClinicalTrials.gov, and WHO ICTRP without language restrictions. Randomized controlled trials (RCTs) comparing secretagogues to artificial tears were eligible. Data extraction and synthesis were conducted using Covidence and the Cochrane RoB 2 tool, and 19 RCTs (n = 2697) were included. Fifteen were analyzed quantitatively; however, only eight trials evaluating diquafosol were suitable for meta-analysis, as data for pilocarpine and cevimeline were insufficient for quantitative synthesis. GRADE was used to assess evidence certainty. PROSPERO registration: CRD42020218407. Diquafosol significantly improved rose bengal staining at 4 weeks and OSDI scores and TBUT in post-cataract patients at 4 and 12 weeks. However, it increased mild adverse events (RR, 1.81; 95% CI, 1.15–2.84). Evidence for pilocarpine and cevimeline was limited. Diquafosol 3% shows greater efficacy than artificial tears in post-cataract DED but with more side effects. Further research is needed for other secretagogues. Full article
(This article belongs to the Special Issue Molecular Advances in Dry Eye Syndrome)
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6 pages, 2701 KiB  
Case Report
Corneal Edema from Accidental Instillation of Stamper Ink Mistaken for Artificial Tears: A Case Report
by Lily S. Ardiani, Sharita R. Siregar and Iwan Soebijantoro
BioMed 2025, 5(3), 18; https://doi.org/10.3390/biomed5030018 - 22 Aug 2025
Abstract
Background: The incidence of chemical ocular trauma after accidentally instilling the “wrong” eyedrops is still frequent, but cases resulting from stamper ink refills are rare. Case Presentation: A 73-year-old man presented to our emergency department with a history of inadvertently instilling stamper ink [...] Read more.
Background: The incidence of chemical ocular trauma after accidentally instilling the “wrong” eyedrops is still frequent, but cases resulting from stamper ink refills are rare. Case Presentation: A 73-year-old man presented to our emergency department with a history of inadvertently instilling stamper ink refill into both eyes (BEs) instead of artificial tears. Immediate irrigation and evaluation were performed. The initial visual acuity (VA) was 0.4 in the right eye (RE) and 0.8 in the left eye (LE). Slit lamp examination showed edema palpebra with periocular blue staining in BEs, chemotic conjunctiva with a much darker color in the RE than the LE, and epithelial defects with a positive fluorescein test in BEs. A diagnosis of bilateral corneal abrasion and chemotic conjunctiva was established. Ten hours after the emergency visit, RE VA decreased to 0.2, and corneal edema was found during the follow-up examination. Medications including levofloxacin antibiotic, sodium hyaluronate, sodium chloride, combined polymyxin sulfate–neomycin sulfate and dexamethasone eyedrops, mefenamic acid, and ascorbic acid tablets were prescribed. The RE corneal edema still occurred, and the endothelial cell count was 1952 and 987 cells/mm2 in the RE and LE at the one-week follow-up. After three weeks, corneal edema had fully resolved, and the VA was 0.4 and 0.8 in the RE and LE, respectively. Conclusions: This case report adds to the spectrum of the continuing problem of chemical ocular trauma after mistakenly instilling the eyedrops. Promoting and changing to different packages for non-ophthalmic products in plastic bottles mimicking eyedroppers is essential to minimize these injuries. Full article
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16 pages, 6764 KiB  
Article
Hepatocellular Early Apoptosis Associated with HES 130/0.4 Administration for Volume Replacement in Pigs After Severe Bleeding
by Helena Vala, Ana I. Faustino-Rocha, Rita Cruz, Carlos Venâncio, Aura Silva, João R. Mesquita, Ana Liza Ortiz and David A. Ferreira
Vet. Sci. 2025, 12(9), 787; https://doi.org/10.3390/vetsci12090787 - 22 Aug 2025
Abstract
Hydroxyethyl starch (HES) 130/0.4 is commonly used for volume replacement, yet its hepatic effects in the context of acute haemorrhage remain unclear. This study aimed to evaluate hepatic histopathological changes related to HES 130/0.4 administration when compared to Ringer’s lactate (RL) in healthy [...] Read more.
Hydroxyethyl starch (HES) 130/0.4 is commonly used for volume replacement, yet its hepatic effects in the context of acute haemorrhage remain unclear. This study aimed to evaluate hepatic histopathological changes related to HES 130/0.4 administration when compared to Ringer’s lactate (RL) in healthy pigs subjected to acute bleeding under general anaesthesia. Eighteen pigs were randomised into three groups: RL (n = 6), HES 130/0.4 (n = 6), and a non-bleeding control (n = 6). Liver tissue was collected postmortem and analysed using haematoxylin–eosin staining, cytochrome c immunohistochemistry, the TUNEL assay, and M30 immunofluorescence. No statistically significant differences were observed in general histopathological changes, TUNEL, or cytochrome c expression (p > 0.050). However, the pigs that received HES 130/0.4 for volume replacement showed significantly higher intensity of the liver M30 immunostaining in the Q-score (p < 0.010), H-score (p < 0.010), and c indexc index (p < 0.050) when compared to animals that received Ringer’s lactate solution or animals in the control group. These findings suggest that HES 130/0.4 induces increased early hepatocellular apoptosis when compared to RL in this model, raising concerns about its hepatic safety profile under haemorrhagic conditions. Full article
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22 pages, 11866 KiB  
Article
Study on the Mechanism of RuHaoDaShi Granules in Treating H1N1 Viral Pneumonia Based on Network Pharmacology and Experimental Validation
by Aixin Chen, Tianhang Chen, Yu He, Jiehong Yang and Haitong Wan
Pathogens 2025, 14(8), 834; https://doi.org/10.3390/pathogens14080834 - 21 Aug 2025
Abstract
Objective: This study aims to investigate the pharmacodynamic effects and underlying mechanisms of the Chinese herbal formula RuHaoDaShi (RHDS) granules against the influenza virus in experimental models. Methods: This study aims to employ network pharmacology to identify the active components of RHDS and [...] Read more.
Objective: This study aims to investigate the pharmacodynamic effects and underlying mechanisms of the Chinese herbal formula RuHaoDaShi (RHDS) granules against the influenza virus in experimental models. Methods: This study aims to employ network pharmacology to identify the active components of RHDS and its potential targets and mechanisms of action against H1N1. The molecular docking approach validated the interactions between the core targets and the RHDS compounds. In vitro, the antiviral activity of RHDS was assessed by therapeutic, prophylactic, and premixed administration to H1N1-infected A549 cells. An in vivo experiment was conducted using a mouse H1N1 pneumonia model. The model was treated with a dose of 1.04, 2.08, and 4.16 g/kg of RHDS, administered via gavage daily. The study’s objective was to evaluate the antiviral activity and mechanism of action of RHDS in mice. Mice were evaluated on day 6 by assessing survival, viral load (RT-qPCR), lung pathology (HE staining), inflammatory cytokines (ELISA, immunohistochemistry), and ferroptosis markers (WB, qPCR). Results: Network pharmacology identified 77 biologically active RHDS compounds (e.g., quercetin and kaempferol) and 32 core targets common to RHDS, H1N1, and ferroptosis. Molecular docking was used to verify a high affinity for binding between the core targets HIF-1α, MAPK3, and key RHDS compounds. In vitro studies demonstrated that RHDS exhibited protective properties against H1N1-infected cells, with the therapeutic delivery method proving the most efficacious. In vivo studies have shown that RHDS reduces mortality, lung index, and viral load in mice while attenuating histopathological damage. The study demonstrated a reduction in the release of inflammatory cytokines, including IL-6, IFN-γ, and IL-17A, and decreased expression levels of MPO and F4/80 proteins in lung tissue. Mechanistically, the administration of RHDS resulted in the up-regulation of the expression levels of GPX4, SLC7A11, and Nrf2 proteins while concomitantly inhibiting the expression of HIF-1α, COX2, and ACSL4. These findings confirm the modulatory effect of RHDS on the GPX4/SLC7A11/Nrf2 pathway. Conclusions: RHDS demonstrated a protective effect against H1N1-induced cytopathy in vitro and was effective in attenuating H1N1-induced pneumonia in murine models. The study suggests that RHDS has antiviral potential to treat H1N1 viral pneumonia by modulating inflammatory cytokines and the GPX4/SLC7A11/Nrf2 pathway. Full article
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20 pages, 3581 KiB  
Article
Long-Term Durability and Variant-Specific Modulation of SARS-CoV-2 Humoral and Cellular Immunity over Two Years
by Lilia Matei, Mihaela Chivu-Economescu, Laura Denisa Dragu, Camelia Grancea, Coralia Bleotu, Raluca Hrișcă, Corneliu Petru Popescu, Carmen C. Diaconu and Simona Maria Ruţă
Int. J. Mol. Sci. 2025, 26(16), 8106; https://doi.org/10.3390/ijms26168106 - 21 Aug 2025
Abstract
There is an increasing need to understand the long-term dynamics and quality of SARS-CoV-2 immune memory—both humoral and cellular—particularly with emerging variants. This study aimed to evaluate immune durability and variant-specific modulation through a longitudinal analysis of individuals with diverse SARS-CoV-2 exposure histories, [...] Read more.
There is an increasing need to understand the long-term dynamics and quality of SARS-CoV-2 immune memory—both humoral and cellular—particularly with emerging variants. This study aimed to evaluate immune durability and variant-specific modulation through a longitudinal analysis of individuals with diverse SARS-CoV-2 exposure histories, over two years after infection and/or vaccination. The study involved assessing anti-spike IgG and IgA levels over time and analyzing their relationship with neutralizing activity against both ancestral and Omicron SARS-CoV-2 variants. Persistence of T cell responses was evaluated using intracellular cytokine staining (ICS) and activation-induced marker (AIM) assays. Anti-S IgG levels remained stable over time and increased after each immune stimulation, suggesting cumulative immune memory. Neutralizing capacity correlated strongly with IgG levels, showing long-term stability for pre-Omicron variants, but a moderate decline for Omicron. CD4+ and CD8+ T cell responses persisted across all groups, largely unaffected by Omicron mutations. However, cytokine profiles revealed subtle, variant-dependent changes. These findings underscore the durability of cellular immunity and the comparatively reduced robustness of Omicron-specific humoral responses. Such insights are crucial for understanding long-term protection against evolving SARS-CoV-2 variants and guiding public health strategies. Full article
(This article belongs to the Special Issue COVID-19: Molecular Research and Novel Therapy)
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12 pages, 2542 KiB  
Article
Cumulative Low-Dose-Rate Radiation Induces Oxidative Stress, Apoptosis, and Fibrosis in Mouse Testis
by Eun-Jin Kim, Anjas Happy Prayoga, Jina Ha, Deok Gyeong Kang, Jinsung Yang, Sohi Kang, Jin-Mok Kim, Byeonggyu Ahn, Dang Long Cao, Seung Pil Yun, Bo Hyun Lee, Joong-Sun Kim and Dawon Kang
Antioxidants 2025, 14(8), 1028; https://doi.org/10.3390/antiox14081028 - 21 Aug 2025
Abstract
Ionizing radiation is a well-known environmental stressor capable of generating excessive reactive oxygen species (ROS), leading to oxidative damage in sensitive tissues, including the reproductive system. While oxidative stress is increasingly implicated in male reproductive dysfunction, the long-term effects of low-dose-rate (LDR) radiation [...] Read more.
Ionizing radiation is a well-known environmental stressor capable of generating excessive reactive oxygen species (ROS), leading to oxidative damage in sensitive tissues, including the reproductive system. While oxidative stress is increasingly implicated in male reproductive dysfunction, the long-term effects of low-dose-rate (LDR) radiation on testicular structure and oxidative status remain underexplored. In this study, mice were exposed to continuous LDR radiation (0.39, 1.29, and 3.46 mGy/h) for 21 days to assess testicular histopathology and oxidative status. Although testis weight did not significantly differ among groups, histological analysis revealed basal membrane disruption and reduced spermatogenic cell populations in irradiated groups. Masson’s Trichrome and Sirius Red staining demonstrated dose-dependent collagen deposition, indicating progressive testicular fibrosis. TUNEL assays confirmed increased germ cell apoptosis in the mid- and high-dose-rate groups. ROS levels were significantly elevated only in the highest-dose group, suggesting a threshold-dependent oxidative stress response. These findings indicate that chronic LDR radiation induces testicular damage primarily through apoptosis and fibrosis, with oxidative stress potentially contributing at higher exposure levels. Full article
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