Search Results (7,689)

Currently, the main method for producing the vitamin C precursor 2-keto-l-gulonic acid (2-KLG) is a two-step fermentation process, in which secondary sterilization and fermentation processes result in higher costs and energy consumption. Consequently, the development of a one-step fermentation process is seen as a more desirable approach for 2-KLG production. In this study, we used Gluconobacter oxydans WTF0512 and Ketogulonicigenium vulgare WTF0114 as co-cultured strains for the production of 2-KLG from d-sorbitol via one-step fermentation. The fermentation behaviors of G. oxydans WTF0512 and K. vulgare WTF0114 were initially investigated. Subsequently, the fermentation process and medium were optimized, and the titer of 2-KLG reached 132.99 ± 0.52 g/L, with a molar conversion rate of 92.42%, which, to the best of our knowledge, is the highest production via one-step fermentation reported to date. These findings will provide a basis for developing a more economical large-scale one-step fermentation process for the production of 2-KLG. Full article

Chondroitin sulfate (CS) chains are major components of the extra- and pericellular matrix in the central nervous system (CNS), and their sulfation patterns influence CNS development and function. Highly sulfated CS preparations, including CS-D- and CS-E-enriched forms, have been shown to facilitate neurite outgrowth in cultured mouse hippocampal neurons. Notably, neurons cultured on CS-D- or CS-E-enriched substrates exhibited the following distinct morphological characteristics: CS-D promoted the extension of multiple short neurites, whereas CS-E induced the formation of a single elongated neurite with a polarization-like morphology. These features are consistent with early stages of neuronal polarization. However, the specific roles of these highly sulfated CS forms in polarization-like morphology remain unclear. In this study, we demonstrate that polarization-like morphological transitions in hippocampal neurons can be modulated on mixed CS-D/CS-E substrates by varying their ratios. Compared with CS-D-enriched substrates, CS-E-enriched substrates more effectively promoted polarization-like neuronal morphology, accompanied by enhanced activation of Src-family kinases. Furthermore, forced activation of Fyn kinase induced morphological changes resembling polarization-like features in a neuroblastoma cell line, even in the absence of CS-D/CS-E mixed substrates. In conclusion, highly sulfated CS subtypes may function as extracellular cues that regulate neuronal morphology via Src-family signaling, with likely involvement of Fyn. Full article

Ledum palustre (L. palustre) is widely used in drug development because of its antibacterial and analgesic effects. However, wild L. palustre is often affected by wildfires, resulting in unstable yields. Forest fires represent a major disturbance in northern forest ecosystems and profoundly affect shrub vegetation and its associated rhizosphere microbial communities. In this study, we investigated a fire chronosequence (1991, 2004, 2012, 2017, and 2020) to systematically examine the morphological traits of L. palustre, rhizosphere soil physicochemical properties, and microbial community characteristics and to identify the key drivers underlying these patterns. The results revealed that postfire recovery time significantly influenced the morphological traits of L. palustre. The biomass, branch number, basal diameter, and plant height of the shrubs at the 1991 burned site increased by 270.49%, 36.11%, 79.32%, and 191.36%, respectively (p < 0.05). From unburned soils, 29 bacterial and 29 fungal isolates were obtained, with Bacillus sp. and Oidiodendron sp. being the dominant culturable bacterial and fungal taxa, respectively. With increasing postfire recovery time, soil moisture, total nitrogen, ammonium, nitrate, soil organic carbon, acid phosphatase (AP) and N-acetyl-β-D-glucosaminidase (NAG) activity significantly decreased. Early fire disturbance markedly altered soil microbial abundance and community composition, leading to an overall decrease in bacterial α diversity. The bacterial community structure at the 2020 burn site and the fungal community structure at the 2012 burn site significantly differed. Mantel tests revealed significant positive correlations between branch number and basal diameter (p < 0.01) and significant negative correlations between plant height and stem density (p < 0.001). Soil carbon and hydrolysable nitrogen were significantly positively correlated with AP and NAG activities (p < 0.001). Moreover, soil physicochemical properties significantly shaped soil microbial community structures, with bacterial communities in early postfire sites driven by total carbon and nitrogen (p < 0.05), whereas fungal communities in the 2012 burned site were influenced primarily by β-N-acetylglucosaminidase (BG) activity (p < 0.05). Fire disturbance drives successional changes in the rhizosphere microbial community structure and function by altering the soil nutrient status and enzyme activity, which in turn influences the morphological traits of L. palustre. This study provides a theoretical basis for improving the yield of L. palustre by exploring the variation in rhizosphere microorganisms. Full article

Background/Objectives: The Black Caribbean population of Honduras, also referred to locally as Negro Inglés, constitutes one of the country’s nine recognized indigenous and Afro-descendant peoples. Predominantly settled in the Bay Islands and sections of the Caribbean coast, this community traces its ancestry predominantly to West Africa and has remained culturally and linguistically distinct for more than three centuries. Despite its demographic and historical relevance, no population-specific short tandem repeat (STR) database has been established for this group. Methods: Allele frequencies for 23 autosomal STR loci were characterized in 100 unrelated Black Caribbean individuals from the department of Islas de la Bahía. DNA was extracted from blood on FTA cards and amplified with the PowerPlex Fusion 6C System (Promega Corporation). Statistical parameters were computed using Genepop v4.2, Arlequin v3.5 and GDA v1.0. Results: A total of 241 distinct alleles were detected across all 23 loci (mean 10.48 ± 3.85 alleles/locus). Expected heterozygosity ranged from 0.6541 (D13S317) to 0.9350 (SE33), with a mean of 0.8150 ± 0.0664—values consistent with a population of predominantly West African origin. No locus exhibited a significant departure from Hardy–Weinberg equilibrium after Bonferroni correction (α = 0.0022). The combined power of discrimination exceeded 99.9999% and the combined chance of exclusion surpassed 99.9999%. Conclusions: This first genetic characterization of the Honduran Black Caribbean population delivers an essential, population-specific reference dataset for forensic casework, paternity testing, and population genetics research. The data also deepen the understanding of Afro-descendant genetic diversity in Central America and constitute a critical step towards equitable forensic genetic services for all Honduran ethnic communities. Full article

High-quality genomic DNA extraction remains a major bottleneck for fungal genomics, particularly for worldwide aerobic and non-photosynthetic mushroom species that rely on their rigid cell walls, interference between metabolites, polysaccharides, etc., and complex genomes. This study systematically compares five DNA extraction protocols involving four distinct sample preparation procedures (fresh (A), filtered (B), frozen (C) and cryogenic mycelium (D)) across mycelial cultures of eight Tunisian fungal strains representing Ascomycota and Basidiomycota to identify the optimal combination for genomic DNA extraction from mycelium. The eight phylogenetically diverse fungal species were analyzed using short-read (MiSeq and NextSeq550) and/or long-read (MinION Mk1C) sequencing technologies, giving a depth coverage between 3.7× and 83×. The generation and quality of the assemblies were assessed within the Galaxy platform, which revealed a gap percentage of 0–0.509%. Taxonomic characterization and phylogenetic inference were performed with SANGER technology using the Internal Transcribed Spacer (ITS) and D1/D2 region of the 26S rRNA gene, assigning the species to our eight different strains: Clitopilus baronii (BS6), Porostereum spadiceum (BS200), Trametes versicolor (BS22-9), Schizophyllum commune (BS23-13), Gloeophyllum abietinum (BS23-14), Irpex laceratus (BS100), Trichoderma asperellum (GC9) and Trichoderma harzianum (S3). The optimized DNeasy Plant Pro Kit protocol with cryogenic biomass treatment presents a safe and cost-effective method for fungal genome sequencing and taxonomic resolution. This integrated comparative evaluation of extraction for sequencing identifies an optimal Qiagen-based extraction strategy combined with cryogenic treatment for eight diverse Tunisian fungal species, guiding method selection based on specific cell wall characteristics rather than proposing a universal protocol limited by unequal replication and strain numbers. Full article

Microbial contamination in aquatic environments poses severe threats to aquaculture sustainability, ecological balance and public health. Traditional culture-based detection methods, while standardized, are time-consuming and labor-intensive, often failing to meet the urgent need for rapid on-site monitoring required to prevent disease outbreaks and manage water quality effectively. By integrating latest research advances (2020–2025), this study reviews advances in rapid detection technologies for aquatic microorganisms, including the evolution of nucleic acid amplification strategies, with a focused comparison of the analytical sensitivity and field deployability of quantitative polymerase chain reaction (qPCR) and mainstream isothermal amplification techniques (loop-mediated isothermal amplification, LAMP; recombinase polymerase amplification, RPA). Furthermore, this study reports on the emergence of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated protein (Cas) systems as next-generation diagnostic tools, highlighting their integration with microfluidic Lab-on-a-Chip (LOC) platforms to achieve attomolar sensitivity. We also consider the application of portable nanopore sequencing for real-time pathogen identification and the growing role of Artificial Intelligence (AI) in analyzing complex diagnostic datasets. Advanced molecular methods have achieved significant reductions in time consumption—from days to less than one hour—while challenges regarding sample preparation and environmental matrix inhibition remain. The future of aquatic monitoring lies in integrated, automated systems that combine the specificity of CRISPR-Cas diagnostics with the connectivity of IoT-enabled biosensors. Comparative analysis indicates that isothermal amplification methods (LAMP, RPA) coupled with CRISPR-Cas systems offer the optimal balance of sensitivity, speed, and field deployability for point-of-care aquaculture diagnostics, while qPCR/dPCR remain indispensable for quantitative regulatory applications. We propose a structured technology selection framework to guide researchers and practitioners in choosing appropriate detection modalities based on specific sensitivity, cost, throughput, and deployment requirements. Full article

The hypoxic microenvironment plays a critical role in the progression of canine osteosarcoma (OSA) by promoting different cellular responses, including the release of extracellular vesicles (EVs). Given the clinical aggressiveness of canine OSA, the aim of this study was to evaluate the miRNAome profile in EVs released in vitro by four canine OSA cell lines under hypoxic conditions. In particular, for this study we used two commercial canine osteosarcoma cell lines (D17 and D22) and two primary osteosarcoma cell lines obtained in our laboratory (Penny and Wall). D17, D22, Penny, and Wall cell lines were cultured under normoxic and hypoxic conditions (200 µM CoCl₂) for 24 h. EVs were isolated by size-exclusion chromatography and characterized by nanoparticle tracking analysis and Western blotting. miRNAs extracted from EVs were then sequenced and analyzed using bioinformatics approaches. The most representative miRNAs were identified and validated by qPCR using the miRCURY LNA miRNA PCR assay. miRNome profiling identified 233 miRNAs differentially expressed in EVs across all analyzed cell lines. Among these, 94 miRNAs were detected exclusively under hypoxic conditions. From this subset, 43 miRNAs were selected for further validation by qPCR. The qPCR results showed that miR-222-3p and miR-186-5p were significantly downregulated in the Wall cell line under hypoxia (p ≤ 0.05). TargetScan and pathway enrichment analyses demonstrated that miR-186-5p regulates target genes involved in different cellular processes. In human osteosarcoma, low serum levels of miR-222-3p are associated with poor prognosis, while miR-186-5p is recognized as a key hypoxia-responsive miRNA. Collectively, these results suggest the potential of EV-associated miRNAs as biomarkers in canine OSA and support their relevance in translational and comparative oncology. Full article

Social Virtual Reality (VR) is emerging as a powerful medium for remote social interaction and collaboration, enabling multiple users to share experiences together while apart. Likewise, recent advances in multimedia technologies have proposed strategically combining diverse content formats and introducing interaction techniques for recreating virtual environments and engaging with them, respectively. This study pioneers the joint exploration of Social VR enhanced with holographic communication, multimodal content integration, and advanced interaction methods to deliver realistic and interactive group visits to reconstructed cultural heritage sites, specifically an existing restaurant–museum. The reconstructed space is further augmented with Points of Interest (PoIs), which can be freely visited and dynamically activated to provide rich contextual and historical information about the venue. The proposed technology and scenario have been evaluated objectively and subjectively. Results from objective tests offer relevant insights into the technical requirements, performance metrics (including bandwidth usage and latency), and overall system stability. Results from subjective tests with 22 participant pairs reveal high levels of user satisfaction, particularly in terms of immersion, presence, togetherness, and interaction quality regardless of whether participants acted as Guides (interacting with the VR environment) or Followers (observing and following the Guide’s actions). Beyond demonstrating feasibility, the findings from this study prove, for the first time, how strategically combining multi-user holoportation with multimodal content and role-based interactions can enable guided, collaborative cultural or touristic visits that preserve social presence while supporting rich exploration and contextual learning. Full article

Background: Long-bone non-unions complicated by osteomyelitis remain a major reconstructive and healthcare challenge, particularly in resource-limited settings with a high prevalence of multidrug-resistant (MDR) pathogens. Conventional staged management is associated with a prolonged treatment burden, repeated procedures, and delayed functional recovery. This study evaluated the clinical, radiological, functional, and short-term safety outcomes of a single-stage approach using custom-threaded antibiotic-coated locking nails (TACLNs) in a high-resistance cohort. Methods: This prospective single-center cohort study enrolled 30 adults with osteomyelitis-associated femoral or tibial nonunion at a tertiary hospital in Peshawar, Pakistan. All patients underwent radical debridement and single-stage stabilization with a chest tube mold TACLN loaded with vancomycin and gentamicin, with culture-directed adjunctive antibiotics for resistant organisms. Outcomes were assessed at baseline, Weeks 3 and 6, and Month 6 using inflammatory markers, RUST score, VAS pain, EQ-5D-5L, ASAMI criteria, and return to work or usual activity. No formal sample size calculation was performed, and this study was exploratory in nature. Results: The cohort (mean age 44.9 ± 9.9 years) had a challenging microbiological profile, with 40.0% MDR and 13.3% extensively drug-resistant (XDR) infections. By Month 6, short-term infection control was achieved in 96.7% of patients, with significant reductions in ESR and CRP (both p < 0.001). Radiographic union was achieved in 90.0% of cases at a mean of 18.6 weeks, and the mean RUST score improved from 4.87 to 10.43 at the final follow-up. The VAS pain decreased from 5.23 at week 3 to 0.73 at month 6, EQ-5D-5L improved from 0.39 to 0.84, and 90.0% returned to work or usual activity by month 6. No cement debonding, implant failure, or nephrotoxicity was noted. Conclusions: In this single-arm exploratory cohort, TACLNs were associated with favorable short-term infection control, radiographic union, and functional recovery in osteomyelitis-associated long-bone nonunion, including in an MDR/XDR setting. The independent contribution of the threaded core design cannot be established. Larger multicenter comparative studies with longer follow-ups are needed to confirm the durability and implementation feasibility. Full article

Chemotherapy-induced peripheral neuropathy (CIPN) is a dose-limiting toxicity affecting 30–40% of patients treated with neurotoxic chemotherapy. Sensory symptoms arise from injury to dorsal root ganglion (DRG) neurons and their axons; yet, the underlying mechanisms remain incompletely understood. While human induced pluripotent stem cell (iPSC)-derived sensory neuron (iSN) monolayers have provided mechanistic insight, they lack the three-dimensional architecture and cellular heterogeneity of native DRG tissue. Here, we generated human iPSC-derived DRG organoids (iDRGOs) containing mixed neuronal and peripheral glial populations and established a quantitative neurite outgrowth assay to model chemotherapy-induced neurotoxicity in a 3D context. iDRGOs from three healthy donors were exposed to bortezomib, vincristine, or paclitaxel. All three drugs caused dose-dependent neurite outgrowth impairment without significant short-term changes in organoid size, consistent with early axonal injury. Vincristine reduced MAP2 levels when normalized to total protein, whereas bortezomib and paclitaxel showed divergent microtubule-associated responses compared to monolayer cultures. The developmental stage significantly influenced the baseline neurite outgrowth, highlighting the need for age standardization. These results establish iDRGOs as a physiologically relevant human platform that complements monolayer models for mechanistic studies and therapeutic screening in CIPN. Full article

Brain organoids represent three-dimensional structures that allow for human-specific studies in brain development, pathology and therapeutics. These self-organizing systems, formed through the differentiation of human pluripotent stem cells, can mimic important cellular and molecular events of brain development and therefore serve as a platform for the investigation of neurodevelopmental and neurodegenerative diseases, brain injuries, and tumorigenesis. Although brain organoids show promising perspectives in the study of human physiology, existing brain organoid platforms are hindered by issues of under vascularization, immaturity and protocol variability. Nevertheless, the rapid development of new bioengineering, microfluidic and multi-omics tools and approaches allows us to overcome existing problems and increase the physiological significance of these organoids. Brain organoid transplantation and functional studies further enhance the applications of brain organoids in drug screening, disease modeling and personalized medicine. Here, we provide an overview of recent developments in the field of brain organoid cultures, functional characteristics and translational applications. Full article

Activating PIK3CA mutations occur in approximately 40% of hormone receptor-positive (HR+)/HER2-negative breast cancers and represent a major driver of endocrine resistance. The PI3Kα-selective inhibitor alpelisib, in combination with fulvestrant, significantly improves progression-free survival in patients with PIK3CA-mutant disease, as demonstrated in the SOLAR-1 trial. However, this therapeutic strategy is frequently complicated by treatment-induced hyperglycemia, a metabolic disturbance that promotes oxidative stress, mitochondrial dysfunction, and inflammatory signaling, thereby increasing cardiovascular vulnerability. Sodium–glucose cotransporter-2 (SGLT2) inhibitors have emerged as cardiometabolic modulators with benefits extending beyond glucose lowering. In this study, we used a human cardiomyocyte in vitro model designed to recapitulate the hyperglycemic metabolic milieu observed in breast cancer patients receiving PI3Kα-targeted therapy, to investigate whether the SGLT2 inhibitor dapagliflozin directly protects cardiomyocytes from alpelisib- and fulvestrant-induced injury. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were cultured under hyperglycemic conditions (25 mM glucose) to mimic the metabolic environment associated with PI3Kα inhibitor-induced dysglycemia. Cells were exposed to alpelisib (100 nM) and fulvestrant (100 nM), alone or in combination, in the absence or presence of dapagliflozin (1 μM). Cardiomyocyte viability was assessed using the MTS assay, mitochondrial function by TMRM-based mitochondrial membrane potential (ΔΨm) measurements, and apoptosis by caspase-3 quantification. Cardiomyocyte injury was evaluated by release of cardiac troponin I and heart-type fatty acid binding protein (H-FABP). Lipid peroxidation markers (MDA and 4-HNE) were measured to assess oxidative membrane damage. Intracellular inflammasome-related signaling (NLRP3 and MyD88) and secreted inflammatory mediators (IL-1β, IL-18, IL-6, TNF-α, and CCL2) were quantified by ELISA. Exposure to alpelisib, particularly in combination with fulvestrant, significantly reduced cardiomyocyte viability, induced mitochondrial depolarization, and increased caspase-3-mediated apoptotic signaling. These alterations were accompanied by elevated lipid peroxidation (MDA and 4-HNE) and increased release of cardiac injury biomarkers (troponin I and H-FABP). Alpelisib-based treatments also activated inflammasome-related signaling, as indicated by increased intracellular NLRP3 and MyD88 levels and enhanced secretion of pro-inflammatory mediators (IL-1β, IL-18, IL-6, TNF-α, and CCL2). Co-treatment with dapagliflozin significantly attenuated these alterations, preserving mitochondrial membrane potential, reducing apoptotic signaling, limiting oxidative membrane damage, and suppressing inflammatory cytokine release. This study provides evidence that alpelisib-based therapy under hyperglycemic conditions is associated with oxidative, mitochondrial, and inflammatory stress responses in human cardiomyocytes, recapitulating key features of cardiometabolic stress relevant to PI3Kα-targeted therapy. Importantly, dapagliflozin markedly attenuated these alterations, supporting a potential cardioprotective role that may extend beyond glycemic control. These findings provide a mechanistic rationale for further investigation of SGLT2 inhibition as a cardiometabolic protective strategy in patients receiving PI3Kα inhibitor-based cancer therapy. Full article

Background/Objectives: Peritoneal micrometastases (micromets) remain a major barrier to durable cytoreduction in ovarian and other intra-abdominal cancers because lesions are difficult to visualize and are often resistant to systemic therapy. Liposomal doxorubicin (Dox) improves pharmacokinetics but can be limited by slow intratumoral release. Porphyrin-phospholipid (PoP) liposomes enable near-infrared light–triggered release of Dox (chemophototherapy (CPT)), creating an opportunity for intraoperative fluorescence-guided treatment planning and monitoring. Here, we evaluate a laparoscopic fluorescence imaging platform for quantifying light-triggered drug delivery. Methods: LC-Dox-PoP was applied to SCC2095sc and SKOV-3 cultures in 2D monolayers and 3D spheroid clusters. Dox fluorescence was quantified using a laparoscopic fluorescence imaging system over 1–9 μg/mL concentrations and compared with standard well-plate reader measurements. Porphyrin fluorescence was monitored to assess spheroid localization and photobleaching after activation light exposure. Results: For both cell lines, Dox fluorescence exhibited an approximate 4-fold increase at the maximum administered LC-Dox-PoP concentration, following a linear trend in both SCC2095sc and SKOV-3 cultures (R² = 0.97, 0.98 for 2D and R² = 0.98, 0.98 for spheroids). Laparoscope-derived fluorescence measurements agreed with well-plate reader measurements (R² = 0.89–0.96). Porphyrin fluorescence provided stronger complementary contrast for localizing spheroid constructs and decreased after activation light exposure, consistent with photobleaching during triggered release. Conclusions: These results support a quantitative imaging framework for fluorescence-guided monitoring of light-triggered liposomal drug release and may enable individualized CPT dosimetry for peritoneal micrometastases. Findings in SCC2095sc additionally suggest potential relevance of fluorescence-guided CPT for head and neck/oral cancer, where localized post-resection adjuvant treatment may improve control of residual disease. Full article

Human adipose biology is strongly influenced by three-dimensional (3D) architecture, cell–cell interactions, and local oxygen availability maintained over a long-term culture period, features that are not reproduced in traditional two-dimensional (2D) culture systems. To address this gap, we established a long-term human adipose-derived stem cell (hASC) spheroid model using elastin-like polypeptide–polyethyleneimine (ELP-PEI) coating. The ELP-PEI coating facilitated stable spheroid formation and sustained adipogenic differentiation over 56 days. As spheroids enlarged and matured, they exhibited hallmark features of adipocytes, including lipid accumulation, morphological compaction, and transition out of the proliferative state. Glucose uptake increased during maturation and declined as spheroids became larger. This reduction coincided with a marked rise in hypoxia-inducible factor-1α (HIF-1α) expression, indicating the emergence of a hypoxic microenvironment within larger spheroids. Notably, inhibiting HIF-1α restored insulin-stimulated glucose uptake, demonstrating that hypoxia was the primary driver of impaired insulin responsiveness in late-stage spheroids. These findings position ELP-PEI-supported hASC spheroids as a practical and physiologically relevant platform for studying human adipocyte biology, particularly the development and reversibility of hypoxia-associated metabolic dysfunction. This model offers new opportunities for mechanistic studies and for evaluating therapeutic strategies targeting insulin resistance and adipose tissue pathology. Full article

The accurate interpretation of structural edges requires distinguishing geometry-driven discontinuities from reflectance- and illumination-induced variations. Conventional photogrammetric pipelines rely primarily on radiometric and geometric cues, which often lack physical interpretability under complex material and lighting conditions. This study proposes a photogrammetry–polarimetry fusion framework for physics-guided semantic classification of 3D structural edges. Radiometric, geometric, and polarimetric features are integrated within a noise-normalized representation to enable modality-independent interpretation. A rule-based classification scheme is introduced to assign edges to physically meaningful categories, including geometric, material, specular, illumination, and polarization-driven phenomena. The method is evaluated on a calibrated geometric object and a cultural heritage statue. Results show that polarization provides complementary information that reduces ambiguity between geometry-driven and reflectance-driven edge responses while preserving the underlying reconstructed geometry. On the calibrated dataset, edge detection achieves 88.4% precision, 95.5% recall, and an F1-score of approximately 0.92. Multi-view integration further improves the completeness of geometry-dominant 3D edges. The proposed framework introduces a physics-guided semantic sensing layer for multi-modal 3D perception, enabling more robust and interpretable structural analysis in photogrammetric workflows. Full article