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13782 KB  
Article
A Hybrid Framework for Red Blood Cell Labeling Using Elliptical Fitting, Autoencoding, and Data Augmentation
by Bundasak Angmanee, Surasak Wanram and Amorn Thedsakhulwong
J. Imaging 2025, 11(9), 309; https://doi.org/10.3390/jimaging11090309 (registering DOI) - 9 Sep 2025
Abstract
This study aimed to develop a local dataset of abnormal RBC morphology from confirmed cases of anemia and thalassemia in Thailand, providing a foundation for medical image analysis and future AI-assisted diagnostics. Blood smear samples from six hematological disorders were collected between April [...] Read more.
This study aimed to develop a local dataset of abnormal RBC morphology from confirmed cases of anemia and thalassemia in Thailand, providing a foundation for medical image analysis and future AI-assisted diagnostics. Blood smear samples from six hematological disorders were collected between April and May 2025, with twelve regions of interest segmented into approximately 34,000 single-cell images. To characterize cell variability, a convolutional autoencoder was applied to extract latent features, while ellipse fitting was used to quantify cell geometry. Expert hematologists validated representative clusters to ensure clinical accuracy, and data augmentation was employed to address class imbalance and expand rare morphological types. From the dataset, 14,089 high-quality single-cell images were used to classify RBC morphology into 36 clinically meaningful categories. Unlike existing datasets that rely on limited or curated samples, this dataset reflects population-specific characteristics and morphological diversity relevant to Southeast Asia. The results demonstrate the feasibility of establishing scalable and interpretable datasets that integrate computational methods with expert knowledge. The proposed dataset serves as a robust resource for advancing hematology research and contributes to bridging traditional diagnostics with AI-driven clinical support systems. Full article
(This article belongs to the Section Medical Imaging)
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Article
Fungifuels: Polyethylene Decomposition and Electricity Generation with Aspergillus ochraceopetaliformis in Microbial Fuel Cell Systems
by Rojas-Flores Segundo, Magaly De La Cruz-Noriega, Nancy Soto-Deza, Nélida Milly Otiniano, Cabanillas-Chirinos Luis and Anibal Alviz-Meza
Fermentation 2025, 11(9), 527; https://doi.org/10.3390/fermentation11090527 (registering DOI) - 9 Sep 2025
Abstract
Plastic pollution is an increasingly pressing environmental concern due to its persistence in ecosystems. To address this issue, this study evaluates polyethylene biodegradation and bioelectricity generation using Aspergillus ochraceopetaliformis in microbial fuel cells (MFCs). Single-chamber MFCs were designed (three) with carbon and zinc [...] Read more.
Plastic pollution is an increasingly pressing environmental concern due to its persistence in ecosystems. To address this issue, this study evaluates polyethylene biodegradation and bioelectricity generation using Aspergillus ochraceopetaliformis in microbial fuel cells (MFCs). Single-chamber MFCs were designed (three) with carbon and zinc electrodes, where the fungus was cultivated in a nutrient-rich medium to enhance its metabolic activity. Parameters such as pH, power density, and FTIR spectra were monitored to assess plastic biodegradation. The results demonstrated a significant reduction in polyethylene mass and structure, along with a maximum generation of 0.921 V and 4.441 mA on day 26, with a power density of 0.148 mW/cm2 and a current of 5.847 mA/cm2. The optimal pH for fungal activity in the MFC was recorded at 7.059. Furthermore, FTIR analysis revealed a decrease in peak intensity at 1470 cm−1 and 723 cm−1, indicating structural modifications in the treated plastics. Furthermore, microbial fuel cells connected in series successfully powered an LED bulb, generating a maximum voltage of 2.78 V. These findings confirm the feasibility of using Aspergillus ochraceopetaliformis for biodegradation and bioelectricity generation, although practical applications require further optimization of system conditions and improvements in long-term stability. This research contributes to the development of biotechnological strategies for plastic waste management, sustainable integrating approaches with energy potential. Full article
(This article belongs to the Section Industrial Fermentation)
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Article
Monitoring Sublethal Injury in Listeria monocytogenes During Heat Treatment of Pork Frankfurter-Type Sausages: A Single-Cell vs. Population Level Approach
by Marianna Arvaniti, Eleni Vlachou, Maria Kourteli, Anastasia E. Kapetanakou and Panagiotis N. Skandamis
Foods 2025, 14(17), 3144; https://doi.org/10.3390/foods14173144 (registering DOI) - 8 Sep 2025
Abstract
Listeria monocytogenes is a foodborne pathogen capable of contaminating ready-to-eat meat products, e.g., frankfurters. Post-packaging mild heat treatment via water immersion is commonly employed; however, this may be sublethal to cells located in protected niches or beneath the product surface. The objectives of [...] Read more.
Listeria monocytogenes is a foodborne pathogen capable of contaminating ready-to-eat meat products, e.g., frankfurters. Post-packaging mild heat treatment via water immersion is commonly employed; however, this may be sublethal to cells located in protected niches or beneath the product surface. The objectives of this study were to evaluate thermal injury of L. monocytogenes on frankfurters at single-cell versus population level and to comparatively estimate pathogens’ physiological status. Pork frankfurter-type sausages were inoculated (ca. 7.0‒7.5 log CFU/cm2) with L. monocytogenes strain EGDE-e. Heat treatment was performed at 61°C (max. 60 min) and 64 °C (max. 12 min). To determine the injured subpopulation from the total, tryptic soy agar with 0.6% yeast extract (TSAYE), supplemented or not with 5% NaCl, was used. Plating-based quantification of injured cells was compared to CFDA/PIstained cells analysed by fluorescence microscopy and quantified with Fiji software. Injury was recorded mainly after 2 and 4 min at 64 °C, whereas no injury was detected at 61 °C, at population level. Following exposure to 61 °C for 60 min, culturable cells dropped below the enumeration limit (0.3 log CFU/cm2), while a considerable number of CFDA+/PI and CFDA+/PI+ cells indicated viable-but-non-culturable induction and sublethal injury, respectively. These findings suggest that non-culturability may limit the accuracy of solely culture-based enumeration methods. Full article
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Article
Research on the Internal Flow Characteristics of Single- and Coaxial-Nozzle Ejectors for Hydrogen Recirculation in PEMFC
by Jaewoong Han, Seongjae Won and Jinwook Lee
Appl. Sci. 2025, 15(17), 9845; https://doi.org/10.3390/app15179845 (registering DOI) - 8 Sep 2025
Abstract
Hydrogen proton exchange membrane fuel cells (PEMFCs) are a promising clean energy technology for automotive applications owing to their high efficiency and environmentally friendly characteristics. Efficient hydrogen recirculation is critical for sustaining the PEMFC performance, and ejector-based systems offer a passive, energy-efficient solution. [...] Read more.
Hydrogen proton exchange membrane fuel cells (PEMFCs) are a promising clean energy technology for automotive applications owing to their high efficiency and environmentally friendly characteristics. Efficient hydrogen recirculation is critical for sustaining the PEMFC performance, and ejector-based systems offer a passive, energy-efficient solution. However, traditional ejectors suffer from performance degradation across varying fuel-cell loads owing to their limited adaptability. To address this limitation, this study investigated the internal flow behavior and recirculation performance of single- and coaxial-nozzle ejectors, focusing on the influence of the diameter ratio between the mixing chamber and nozzle throat. Numerical simulations were performed to evaluate the flow structures and recirculation ratios under various operating conditions. The diameter ratio between the mixing chamber and the nozzle throat played a crucial role in determining the flow uniformity and recirculation efficiency. Specifically, lower diameter ratios reduce the recirculation ratio across all operating conditions, whereas higher diameter ratios exhibit diminished performance only under very low power outputs (≤4 bar) but show enhanced performance at medium-to-high outputs. These findings suggest that tailoring the geometric parameters of coaxial-nozzle ejectors can significantly improve hydrogen recirculation adaptability in PEMFC systems, thereby supporting more stable and efficient operation across a wide range of vehicle load conditions. Full article
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Review
Differentiation of Human Pluripotent Cells into Pancreatic Beta Cells for Disease Modeling and Cell Replacement Therapy for Diabetes
by Anna A. Barinova, Alexandra Y. Bogomolova, Alexandra N. Bogomazova, Alyona A. Borisova, Sergey L. Kiselev and Alexandra V. Panova
Int. J. Mol. Sci. 2025, 26(17), 8749; https://doi.org/10.3390/ijms26178749 (registering DOI) - 8 Sep 2025
Abstract
Diabetes mellitus (DM) is a metabolic disease characterized by persistent hyperglycemia, resulting from defects in insulin secretion or impaired insulin action. In cases of severe pancreatic cell dysfunction and deficiency, the primary treatment remains lifelong insulin injections. A potential alternative is allogeneic pancreatic [...] Read more.
Diabetes mellitus (DM) is a metabolic disease characterized by persistent hyperglycemia, resulting from defects in insulin secretion or impaired insulin action. In cases of severe pancreatic cell dysfunction and deficiency, the primary treatment remains lifelong insulin injections. A potential alternative is allogeneic pancreatic cell transplantation from a donor, which can stabilize glucose levels. However, the scarcity of donor material and the risk of immune rejection limit the widespread use of this approach. An alternative solution involves using in vitro-derived insulin-producing cells generated through the differentiation of pluripotent stem cells (PSCs), which could overcome the shortage of transplantable material. Furthermore, patient-specific cells—obtained directly from the patient via reprogramming of blood or skin cells into induced pluripotent stem cells (iPSCs)—would avoid immune rejection. Advances in this field have led to the active development and optimization of PSC differentiation into hormone-producing cells worldwide, with more than hundred patients dosed in clinical trials with ESC-derived cells and the single trial of iPSC-derived cells. This review highlights recent progress and prospects in generating insulin-producing cells from human PSCs, their applications in therapy development and disease modeling, as well as the current challenges and potential solutions. Full article
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Article
Ultra-Short-Term Corneal Changes to Nd:YAG Laser Capsulotomy: Energy-Dependent Changes Assessed by Specular Microscopy and Topographic Analysis
by Çağrı Mutaf, Ali Hakim Reyhan, Mübeccel Bulut and Funda Yüksekyayla
Diagnostics 2025, 15(17), 2280; https://doi.org/10.3390/diagnostics15172280 (registering DOI) - 8 Sep 2025
Abstract
Background: This prospective observational study was conducted to systematically assess immediate changes occurring (within one hour) in corneal endothelial cell morphology and anterior segment parameters following Nd:YAG laser posterior capsulotomy in pseudophakic patients and to analyze the correlation between these changes and laser [...] Read more.
Background: This prospective observational study was conducted to systematically assess immediate changes occurring (within one hour) in corneal endothelial cell morphology and anterior segment parameters following Nd:YAG laser posterior capsulotomy in pseudophakic patients and to analyze the correlation between these changes and laser energy parameters. Methods: A single-arm, within-subject pre–post design was employed to evaluate corneal endothelial morphology (cell density, count, area, coefficient of variation and hexagonal percentage) and anterior chamber parameters (depth, angle, volume) before and one hour after the procedure using specular microscopy and Pentacam analysis. Patient demographics (age), clinical parameters (best corrected visual acuity and intraocular pressure), postoperative-YAG laser interval, and laser energy parameters (energy per shot, pulse count, and total applied energy) were also documented. Results: Thirty-two pseudophakic patients (mean age 56.3 ± 19.2 years) underwent Nd:YAG laser posterior capsulotomy with mean energy per shot of 3.15 ± 1.07 mJ and pulse count of 34.3 ± 20.4. Specular microscopy revealed significant post-procedural decreases in endothelial cell density (2184.05 to 2057.2 cells/mm2; p = 0.006) and increases in average cell area (529.25 ± 242.72 to 587.75 ± 281.09 µm2; p = 0.004) and minimum cell area (199.3 ± 170.62 to 248.35 ± 202.7 µm2; p = 0.035). Corneal topography also decreased significantly in the anterior chamber angle (40.07 ± 10.34 to 35.42 ± 6.78 degrees; p = 0.048), with positive correlations between energy per shot and endothelial cell density (r = 0.557; p = 0.011) and average cell area (r = 0.544; p = 0.013). Conclusions: This study demonstrates that Nd:YAG laser capsulotomy causes immediate, energy-dependent alterations in corneal endothelial density and anterior chamber parameters within one hour post-procedurally. The identification of energy per shot as a key determinant represents a preliminary observation for optimizing laser parameters and reducing potential complications in pseudophakic patients. Full article
(This article belongs to the Section Clinical Diagnosis and Prognosis)
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Article
Nested Melanoma and Superficial Spreading Melanoma with Prominent Nests—A Retrospective Study on Clinical Characteristics and PRAME Expression
by Daniela Lenders, Valentin Aebischer, Maximilian Gassenmaier, Matthias Hahn, Gisela Metzler and Stephan Forchhammer
Diagnostics 2025, 15(17), 2279; https://doi.org/10.3390/diagnostics15172279 (registering DOI) - 8 Sep 2025
Abstract
Background: Nested melanoma is a rare subtype of superficial spreading melanoma. Due to its typical histology, characterized by the predominance of large melanocytic nests in an extensive horizontal spread, it is challenging to distinguish it from other benign nested melanocytic lesions. There is [...] Read more.
Background: Nested melanoma is a rare subtype of superficial spreading melanoma. Due to its typical histology, characterized by the predominance of large melanocytic nests in an extensive horizontal spread, it is challenging to distinguish it from other benign nested melanocytic lesions. There is a need to identify additional histopathological parameters that can support the diagnosis of nested melanoma. Methods: In this retrospective case-control study, we analyzed immunohistochemical staining for PRAME in 10 cases of superficial spreading melanoma with prominent nests and 26 nested melanomas. Dysplastic melanocytic nevi were used as the control group. Results: We found a diffuse PRAME positivity (>75% of melanocytes) in 60% of superficial spreading melanoma with prominent nests and 19% of nested melanoma cases, whereas the control group showed no diffuse PRAME positivity. Furthermore, using Melan A immunohistochemistry, we found an absence of pagetoid spread in 31% of nested melanoma and single cells in suprabasal epidermal layers in 69% of cases. All cases with no pagetoid spread were PRAME negative, whereas 28% of cases with a mild pagetoid spread demonstrated diffuse PRAME positivity. Conclusions: We found lower PRAME positivity in nested melanoma compared to superficial spreading melanoma with prominent nests. Particularly in cases without pagetoid intraepidermal spread of melanocytes, negative PRAME staining does not rule out the possibility of nested melanoma. The diagnosis should be made based on typical histomorphological and clinical criteria. Full article
(This article belongs to the Section Pathology and Molecular Diagnostics)
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Article
Risk Factors and Development of a Predictive Model for In-Hospital Mortality in Hemodynamically Stable Older Adults with Urinary Tract Infection
by Tzu-Heng Cheng, Wei Lu, Chen-Bin Chen, Chen-June Seak and Chieh-Ching Yen
Medicina 2025, 61(9), 1625; https://doi.org/10.3390/medicina61091625 (registering DOI) - 8 Sep 2025
Abstract
Background and Objectives: Urinary tract infections (UTIs) are a major cause of emergency department (ED) visits and hospital admissions among older adults. Although most seniors present hemodynamically stable, a sizeable fraction deteriorate during hospitalization, and no ED-specific tool exists to identify those [...] Read more.
Background and Objectives: Urinary tract infections (UTIs) are a major cause of emergency department (ED) visits and hospital admissions among older adults. Although most seniors present hemodynamically stable, a sizeable fraction deteriorate during hospitalization, and no ED-specific tool exists to identify those at greatest risk. We sought to determine risk factors for in-hospital mortality in this population and to develop a predictive model. Materials and Methods: We analyzed the MIMIC-IV-ED database (2011–2019) and enrolled culture-confirmed UTI patients aged ≥ 65 years who were hemodynamically stable—defined as a systolic blood pressure ≥ 100 mm Hg without vasopressor support. Demographics, comorbidities, triage vital signs, and initial laboratory tests were extracted. Least Absolute Shrinkage and Selection Operator (LASSO) regression with 10-fold cross-validation was performed for variable selection. Discrimination was quantified with the C-statistic, calibration with the Hosmer–Lemeshow test, and clinical utility with decision curve analysis. Internal validation was assessed via 1000-sample bootstrap resampling. Results: Among 1571 eligible encounters (median age 79 years, 33% male), in-hospital mortality was 4.5%. LASSO selected eight variables; six remained significant in multivariable analysis: age, systolic blood pressure, oxygen saturation, white blood cell count, red cell distribution width, and blood urea nitrogen. The predictive nomogram demonstrated a C-statistic of 0.73 (95% CI 0.66–0.79) and outperformed traditional early warning scores. Conclusions: A six-variable nomogram may stratify mortality risk in hemodynamically stable older adults with UTI. Because the model was developed in a single U.S. tertiary-care ED, it remains hypothesis-generating until validated in external, multicenter cohorts to confirm generalizability. Full article
(This article belongs to the Section Urology & Nephrology)
13 pages, 324 KB  
Review
Acute and Chronic Immunological Responses to Different Exercise Modalities: A Narrative Review
by Ebru Sever, Sıla Yılmaz and Mitat Koz
Healthcare 2025, 13(17), 2244; https://doi.org/10.3390/healthcare13172244 - 8 Sep 2025
Abstract
The relationship between exercise and immune function has been widely studied, yet findings remain inconsistent regarding how different exercise modalities and intensities influence acute and chronic immunological responses. Previous reviews have often focused on single exercise types or limited outcomes, leaving a gap [...] Read more.
The relationship between exercise and immune function has been widely studied, yet findings remain inconsistent regarding how different exercise modalities and intensities influence acute and chronic immunological responses. Previous reviews have often focused on single exercise types or limited outcomes, leaving a gap for an integrated synthesis. This narrative review aims to address this gap by summarizing and comparing immunological effects across aerobic exercise, resistance training, high-intensity interval training (HIIT), blood flow restriction (BFR), isometric exercise, mind–body interventions, and hypoxic training. A structured narrative approach was adopted. Literature published between January 2000 and December 2024 was searched in PubMed, Scopus, and Web of Science. Experimental and observational studies on humans and animal models were included, with study selection and data extraction performed by two reviewers. Findings were synthesized thematically by exercise modality to capture both acute and chronic immune responses. Twenty-four eligible studies were identified. Aerobic and mind–body exercises consistently demonstrated anti-inflammatory and immunoprotective effects, including increased IL-10 production, improved T cell profiles, and reduced inflammatory markers. Isometric training showed favorable modulation of cytokines and T cell balance, while resistance training evidence was limited but suggested cortisol-lowering benefits. HIIT, BFR, and hypoxic exercise produced mixed results, often combining transient pro-inflammatory responses with immunological benefits. Acute and chronic immunological responses to exercise are highly modality- and intensity-dependent. Aerobic and mind–body interventions provide the most consistent benefits, whereas HIIT, BFR, and hypoxic training show variable effects. Further high-quality trials are needed to clarify mechanisms and guide exercise-based immune recommendations. Full article
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14 pages, 2938 KB  
Article
Genomic and Phylogenomic Characterization of Three Novel Corynebacterium Species from Camels: Insights into Resistome, Mobilome Virulence, and Biochemical Traits
by Haitham Elbir
Microorganisms 2025, 13(9), 2090; https://doi.org/10.3390/microorganisms13092090 - 8 Sep 2025
Abstract
The genus Corynebacterium is commonly isolated from camel uteri, yet it is rarely identified to the species level. During our routine clinical examination of she-camels brought to the hospital with history of reproductive and systemic health issues, four isolates from the uterus and [...] Read more.
The genus Corynebacterium is commonly isolated from camel uteri, yet it is rarely identified to the species level. During our routine clinical examination of she-camels brought to the hospital with history of reproductive and systemic health issues, four isolates from the uterus and one isolate from blood could not be assigned to any valid Corynebacterium species. Therefore, we aim to identify these isolates, determine any potential virulence factors, and describe how gene turnover contributed to the evolution of these species. Genome-based and phenotypic identification, along with resistome, mobilome, virulome and phylogenomics analysis, was used to characterize the isolates. The isolates were Gram stain-positive, catalase-positive, and rod-shaped. The isolates were assigned to the genus Corynebacterium based on 16S rRNA gene sequence similarity and phylogenetic analysis. The isolates 3274 and ayman were classified as two new Corynebacterium species based on the average nucleotide identity (ANI) values of 78.46% and 68.88% and digital DNA–DNA hybridization (dDDH) values of 20.9% and 22.4%. The isolates 2581A, 2583C, and 4168A constitute a single Corynebacterium species based on their pairwise ANI value of 99% and dDDH value of more than 90%. In addition, isolates 2581A, 2583C, and 4168A showed ANI values of 75.99%, 75.86%, and 76.04% and dDDH values of 23.1%, 23%, and 22.5% with closely related species, and were designated as single new Corynebacterium species. Genes for mycolic acid and menaquinone biosynthesis were detected in all isolates. The isolates were susceptible to ceftiofur, linezolid, penicillin, erythromycin, and tetracycline. All isolates harbored the antiseptic resistance gene qacA. Moreover, virulence factors involved in cell adhesion and iron acquisition were detected. The evolution of these species is dominated by gene gain rather than gene loss. The majority of these genes are acquired through horizontal gene transfer, mediated by prophages and genomic islands. In summary, we characterized three new Corynebacterium species, expanding the number of new Corynebacterium species from animals. Moreover, we described the mechanism underlying the genome evolution of these new species. The clinical findings and detection of virulence genes highlight the significance of these isolates as possible pathogens, contributing to the development of endometritis in camels. Full article
(This article belongs to the Special Issue Epidemiology of Zoonotic Pathogens)
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35 pages, 2282 KB  
Review
Cancer-Associated Fibroblasts in Solid Tumors and Sarcomas: Heterogeneity, Function, and Therapeutic Implications
by Omar Badran, Idan Cohen and Gil Bar-Sela
Cells 2025, 14(17), 1398; https://doi.org/10.3390/cells14171398 - 7 Sep 2025
Abstract
Cancer-associated fibroblasts (CAFs) are crucial regulators of the tumor microenvironment (TME), promoting cancer progression, immune suppression, and therapy resistance. Single-cell transcriptomics has identified at least five distinct CAF subtypes: myofibroblastic (myCAFs), inflammatory (iCAFs), antigen-presenting (apCAFs), metabolic (meCAFs), and vascular/developmental (vCAFs/dCAFs), each with unique [...] Read more.
Cancer-associated fibroblasts (CAFs) are crucial regulators of the tumor microenvironment (TME), promoting cancer progression, immune suppression, and therapy resistance. Single-cell transcriptomics has identified at least five distinct CAF subtypes: myofibroblastic (myCAFs), inflammatory (iCAFs), antigen-presenting (apCAFs), metabolic (meCAFs), and vascular/developmental (vCAFs/dCAFs), each with unique localization, signaling, and functions. While CAFs are well studied in epithelial cancers, their roles in sarcomas are less understood despite the shared mesenchymal origin of tumor and stromal cells. This overlap blurs the line between malignant and non-malignant fibroblasts, raising fundamental questions about the identity of CAFs in mesenchymal tumors. In this narrative review, we explore the heterogeneity and plasticity of CAFs across solid tumors, focusing on their role in immune evasion, epithelial-to-mesenchymal transition (EMT), and resistance to chemotherapy, targeted therapy, and immunotherapy. We highlight emerging evidence on CAF-like cells in sarcomas and their contribution to tumor invasion, immune exclusion, and metastatic niche formation. We also assess new strategies to target or reprogram CAFs and suggest that CAF profiling may serve as a potential biomarker for patient stratification. Understanding CAF biology across various tumor types, including those with dense stroma and immunologically cold sarcomas, is crucial for developing more effective, personalized cancer treatments. Full article
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24 pages, 4173 KB  
Article
The Effect of Lurbinectedin as a Monotherapy and in Combination with Ionizing Radiation on Sarcoma Cell Lines
by Hannah Alsheimer, Paula Schiller, Sabine Semrau, Felix Grabenbauer, Rainer Fietkau, Luitpold V. Distel and Laura S. Hildebrand
Cancers 2025, 17(17), 2930; https://doi.org/10.3390/cancers17172930 - 7 Sep 2025
Abstract
Background/Objectives: Soft tissue sarcomas are rare, heterogeneous tumors with limited therapeutic options and suboptimal outcomes in advanced stages. Lurbinectedin is a promising new antineoplastic alkylating agent. This study investigates its cytotoxic effects and its potential as a radiosensitizing agent on soft tissue sarcoma. [...] Read more.
Background/Objectives: Soft tissue sarcomas are rare, heterogeneous tumors with limited therapeutic options and suboptimal outcomes in advanced stages. Lurbinectedin is a promising new antineoplastic alkylating agent. This study investigates its cytotoxic effects and its potential as a radiosensitizing agent on soft tissue sarcoma. Methods: Four soft tissue sarcoma cell lines were treated with lurbinectedin alone or in combination with ionizing radiation. Single-dose irradiation in a 4-day protocol was compared with prolonged treatment and an additional fractionated ionizing radiation scheme in a 6-day protocol. Cellular responses were analyzed by flow cytometry for apoptosis (Annexin V)/necrosis (7AAD) and cell cycle (Hoechst), clonogenic cell survival, and scratch assays for cell migration. Results: In the 4-day protocol, lurbinectedin induced G2/M arrest in all cell lines (p = 0.029) and significantly increased apoptosis/necrosis (p = 0.029) in SW-872. Lurbinectedin-treatment resulted in a decrease (p ≤ 0.002) of clonogenic cells in all cell lines. In the scratch assay, cell migration was delayed in two cell lines (p = 0.048) after lurbinectedin-treatment. Additional radiotherapy had no significant effect compared to lurbinectedin-monotherapy in apoptosis/necrosis and G/2M arrest in the 4-day protocol, clonogenic cell assay, and scratch assay. In the 6-day protocol, lurbinectedin induced an increase (p = 0.029) in G2/M arrest in all cell lines and apoptosis/necrosis in three cell lines, while resulting in a decrease (p < 0.001) of clonogenic cells. Additional radiotherapy had a significant effect on the decrease in clonogenic cells (p ≤ 0.048) in two cell lines but did not increase G2/M arrest and apoptosis/necrosis. Conclusions: Lurbinectedin had strong effects on three of the selected cell lines by inducing G2/M arrest, promoting apoptosis/necrosis, and reducing clonogenic survival, suggesting that it may be a promising chemotherapeutic agent in soft tissue sarcoma treatment. The effect on the fourth cell line was limited, as well as the effect on cell migration. Single-dose irradiation occasionally interfered with the effects of Lurbinectedin, whereas adding fractionated irradiation caused an additional decrease in clonogenic survival, indicating that the combination of Lurbinectedin with fractionated ionizing radiation may have promising effects. Full article
(This article belongs to the Special Issue Radiotherapy for Sarcoma)
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17 pages, 2482 KB  
Article
Identification and Genomic Insights into the Biological Control and Growth-Promoting Mechanism of Bacillus velezensis L11-7, a Potential Biocontrol Agent of Passion Fruit Stem Basal Rot
by Ming Jin, Yuanfeng Tang, Rui Yang, Quan Zeng, Mingxiao Duan, Jieqiu Li and Jiaorong Meng
Microorganisms 2025, 13(9), 2084; https://doi.org/10.3390/microorganisms13092084 - 7 Sep 2025
Abstract
Basal stem rot caused by Fusarium solani is among the most destructive soil-borne diseases affecting passion fruit (Passiflora spp.). While biological control employing antagonistic microorganisms offers a promising plant protection strategy, reports on antagonists specifically targeting passion fruit basal stem rot remain [...] Read more.
Basal stem rot caused by Fusarium solani is among the most destructive soil-borne diseases affecting passion fruit (Passiflora spp.). While biological control employing antagonistic microorganisms offers a promising plant protection strategy, reports on antagonists specifically targeting passion fruit basal stem rot remain limited. Here, a screen for F. solani antagonists led to the identification of Bacillus velezensis strain L11-7, whose whole genome was subsequently sequenced. Pot experiments demonstrated that strain L11-7 significantly reduced the severity of stem basal rot, achieving control efficiencies of 92.85%, and exhibited broad antagonistic properties against other plant pathogenic fungi. L11-7 possesses cellulase, glucanase, and protease activities, alongside capabilities for nitrogen and phosphorus production. L11-7 was identified as B. velezensis through morphological analysis, 16S rRNA, gyrB, and rpoB gene sequencing, and whole-genome analysis. Its genome features a single circular 3.97 Mb chromosome harboring 13 s metabolite biosynthetic gene clusters (e.g., fengycin, surfactin, macrolactin H, bacillaene, difficidin) and genes encoding essential cell wall hydrolases. Several genes related to plant growth promotion, including those involved in nitrogen fixation and IAA production, are also present. These results indicate that B. velezensis L11-7 is a prospective biocontrol agent against passion fruit basal stem rot and has plant growth-promoting properties. Full article
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19 pages, 5339 KB  
Article
The RNA-Binding Protein RBMX Mediates the Immunosuppressive Microenvironment of Osteosarcoma by Regulating CD8+T Cells
by Yu Qiu, Chao Pu, Chengguang Wang and Zhengxue Quan
Cancers 2025, 17(17), 2928; https://doi.org/10.3390/cancers17172928 - 6 Sep 2025
Abstract
Background: The progression of osteosarcoma is closely related to the immune microenvironment. Related studies have found that the RNA-binding motif protein, X-linked (RBMX), plays a regulatory role in modulating the biological characteristics of the tumor microenvironment (TME). However, its regulatory mechanism in osteosarcoma [...] Read more.
Background: The progression of osteosarcoma is closely related to the immune microenvironment. Related studies have found that the RNA-binding motif protein, X-linked (RBMX), plays a regulatory role in modulating the biological characteristics of the tumor microenvironment (TME). However, its regulatory mechanism in osteosarcoma remains unclear. Methods: In this study, the expression of RBMX in osteosarcoma was analyzed using the results of bulk and single-cell transcriptome sequencing of human osteosarcoma. The RBMX knockout cell line was constructed via lentivirus transfection. The mouse subcutaneous implantable tumor model and single-cell transcriptome sequencing analysis revealed the effects of RBMX on the osteosarcoma microenvironment, as verified via multiplex immunofluorescence, flow cytometry, and PCR experiments. Results: Using the TARGET database and multiplex immunofluorescence, we found that RBMX is highly expressed in human osteosarcoma and is associated with poor prognosis. The high expression of RBMX may mediate the immunosuppressive microenvironment of human osteosarcoma. In vitro cell experiments showed that knockout of RBMX significantly inhibited the proliferation of mouse osteosarcoma cells. Through single-cell transcriptome sequencing analysis of subcutaneous implantable tumors in mice, we determined that RBMX deletion substantially elevated the recruitment of cytotoxic CD8+T cells within the mouse TME, which was further verified through flow cytometry analysis. Cell coculture assay confirmed that knockout of RBMX significantly enhanced the cytotoxic activity of CD8+T cells. Finally, cell communication and in vitro experimental verification revealed that knocking out RBMX might enhance the infiltration of CD8+T cells by upregulating histocompatibility 2, K1, and K region (H2-K1) and downregulating thrombospondin 1 (THBS1). Conclusions: This study may provide potential targets for reshaping the immune microenvironment of osteosarcoma and improving its therapeutic efficacy. Full article
43 pages, 4033 KB  
Review
Updated Applications of Stem Cells in Hypoplastic Left Heart Syndrome
by Rui Xiao, Haleema Darr, Zarif Khan and Qingzhong Xiao
Cells 2025, 14(17), 1396; https://doi.org/10.3390/cells14171396 - 6 Sep 2025
Abstract
Hypoplastic left heart syndrome (HLHS) is a severe congenital heart disease affecting 2–3 neonates every 10,000 live births. While prior research has highlighted associations of HLHS with specific chromosomal abnormalities and genetic mutations, the precise pathophysiology remains elusive. Despite early surgical intervention potentially [...] Read more.
Hypoplastic left heart syndrome (HLHS) is a severe congenital heart disease affecting 2–3 neonates every 10,000 live births. While prior research has highlighted associations of HLHS with specific chromosomal abnormalities and genetic mutations, the precise pathophysiology remains elusive. Despite early surgical intervention potentially allowing most HLHS patients to survive their critical heart disease with a single-ventricle physiology, patients frequently experience complications of arrhythmias and right ventricular heart failure, culminating in the need for an eventual heart transplant. Scarcity of suitable donors combined with limited understanding of mechanisms of development highlights the need for furthering our understanding of HLHS and alternative treatment options. Over the past decades, stem cell research has significantly advanced our understanding of cardiac conditions, repair, development, and therapy, opening the door for a new exciting field of regenerative medicine in cardiology with significant implications for HLHS. This review serves to provide a comprehensive overview of a much focused-on area related to HLHS. Specifically, we will first discuss the key pathophysiological basis and signalling molecules of HLHS. We then outline the emerging role of stem cell-based therapy, with a focus on adult stem cells and pluripotent stem cells (PSCs) in uncovering the pathophysiology of HLHS and optimising future treatment directions. Finally, we will also explore the latest and possible future directions of stem cell-derived techniques such as cardiac organoids and bioengineering cardiac tissues and their utility for investigating disease mechanisms, drug screening, and novel therapy for HLHF. Full article
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