Search Results (7,897)

Colon cancer is one of the leading causes of death, requiring advanced therapies that need models for developing new drugs. Conventional cell culture models do not accurately and precisely reproduce the complexity of the tumor microenvironment, limiting their usefulness in research and therapy development. To address this weakness, patient-derived organoids have emerged as promising in vitro models. The implementation of these organoid-based models into more physiologically relevant systems is expected to improve their clinical relevance. Thus, integrating these organoids into microfluidic chips acting as bioreactors will likely improve the predictive response of therapies in personalized medicine. In this article, we report the development of a new colon cancer organoid-on-a-chip model that enables the in vitro culture of patient-derived colon cancer organoids under continuous culture media flow. We demonstrate how the developed organoids were derived from tumor biopsies of patients with colorectal cancer, expanded in standard three-dimensional (3D) culture, and cultured inside the microfluidic chips. The microfluidic chip chambers are designed to house organoids in a controlled environment, allowing the injection of therapies and monitoring by optical microscopy in real time. The in vitro therapies tested were a combination of drugs based on 5-fluorouracil and oxaliplatin at different concentrations. As a result, we demonstrate for the first time that this model proves the capability of this technology for in vitro testing colon cancer therapies. Full article

Cultural Heritage (CH) is becoming increasingly vulnerable to the impacts of climate change, aging, and environmental decay, necessitating advanced preventive conservation strategies. This study presents the results of the SPIDER project, focused on Marquis’s Palace in Botrugno, a small but representative case study in Southern Italy of a municipality overwhelmed with the management of valuable CH sites. The approach integrates multi-sensor surveys, subsurface diagnostics, HBIM modeling, and IoT microclimatic monitoring into a lightweight information model designed for operational flexibility. In addition to that,, the possibility of producing new, eco-friendly filaments for Fused Filament Fabrication (FFF) printing from industrial stone dust waste was explored through a preliminary morphological, structural, and chemical–physical investigation of the stone material historically used in construction, with the aim of identifying materials similar to the original using a simplified, low-cost process. The findings highlight that economic and social factors such as limited resources and the “digital divide” hinder effective technology transfer. Consequently, this study investigates whether a ”lightweight” Asset Information Model (AIM) can provide a more sustainable alternative to complex Digital Twins for small municipalities and other public bodies. For this reason, this research proposes a scalable, wide but basic framework of information management tools and methods aimed at enhancing territorial capacity building, fostering technology integration and social inclusion, and valorizing multidisciplinary approaches to address the challenges affecting CH. Full article

The ubiquitous co-contamination of traditional and novel brominated flame retardants (TBFRs and NBFRs) in anaerobic environments necessitates a comprehensive understanding of their combined environmental fate. This study investigated the anaerobic biotransformation of BDE 99 (a legacy aromatic TBFR) and β-TBCO (a cycloaliphatic NBFR) using a Dehalogenimonas-containing mixed culture (QY2-S1) under single- and co-exposure conditions. In a single-exposure system, QY2-S1 achieved efficient transformation of both substrates, with the observed first-order kinetic constants (k_obs) of 1.31 ± 0.09 d⁻¹ and 2.35 ± 0.13 d⁻¹ for BDE 99 and β-TBCO, respectively. However, the corresponding k_obs values decreased by approximately 3- to 5-fold in a co-exposure system, demonstrating pronounced reciprocal kinetic inhibition. Notably, while culture QY2-S1 maintained a unique ortho-regioselectivity for BDE 99, its stepwise debromination was truncated at the tetra-BDE stage during co-exposure. Absolute quantitative 16S rRNA sequencing identified Dehalogenimonas as the sole organohalide-respiring bacterium, with its abundance initially increasing in tandem with substrate transformation. However, the dehalogenation-coupled cell growth of Dehalogenimonas was significantly modulated by substrate type and combination, with its absolute biomass following the order of: single β-TBCO > co-exposure > single BDE 99. Collectively, these results suggest that the reciprocal kinetic inhibition and the truncation of the BDE 99 debromination pathway under co-exposure are collectively driven by the suppressed growth and substrate preference of Dehalogenimonas. This study provides critical insights for predicting ecological risks and developing bioremediation strategies for co-occurring BFRs in real-world scenarios. Full article

Nutrient supplementation may enhance muscle growth by supporting myogenic activity and regulating metabolic pathways involved in skeletal muscle development. However, the effect of specific nutrient supplementation on satellite cell (SC) activity and muscle-related metabolic pathways remains unclear. Two experiments were conducted to investigate dietary supplements, including guanidinoacetic acid (GAA), 5-hydroxytryptophan (5-HTP), and quercetin, on SC proliferation and differentiation in vitro and on muscle growth and transcriptomics in vivo. For the in vitro study, ovine SC were cultured with different levels (0, 12.5, 25, and 50 μmol/L) of 5-HTP, GAA, or quercetin to examine SC proliferation and differentiation. SC proliferation was greater with 5-HTP and quercetin supplementation, whereas GAA increased SC differentiation. Based on these results, 5-HTP and GAA were selected for the in vivo study because they increased proliferation and differentiation, respectively, and had a wider range of dose effectiveness (12.5 and 25 μmol/L). For the in vivo study, Texel lambs (n = 15; 33 ± 6 kg) were blocked by weight and assigned to one of three treatments: control, GAA, or 5-HTP for 28 d. Supplements were administered sublingually at a dose of 2.5 mg/kg body weight daily before feeding. GAA or 5-HTP did not influence (p > 0.05) average daily gain or dry matter intake. Longissimus muscle area was greater (p < 0.05) at the end of the study compared to the start for all treatment groups. mRNA sequencing showed that GAA supplementation altered gene expression (Padj < 0.05) compared to the control or 5-HTP. However, 5-HTP did not affect (p > 0.05) gene expression compared to the control. LC-MS also showed that plasma 5-hydroxyindoleacetic acid (5-HIAA) was elevated (p < 0.05) in the 5-HTP group. The results suggest that these dietary supplements can stimulate SC proliferation or differentiation in vitro. Short-term supplementation of GAA or 5-HTP in vivo did not alter growth performance, muscle mass, or miR-133a expression; however, GAA supplementation altered the transcriptome to promote amino acid metabolism pathways that are commonly linked to muscle growth through muscle protein synthesis and energy metabolism. Full article

Accurate species-level identification of Legionella during routine environmental surveillance remains challenging, as standard proteomic screening methods like MALDI-TOF MS often lack the resolution to distinguish closely related taxa. This study introduces a novel, cost-effective, and stepwise taxonomic framework that bridges routine environmental monitoring with multi-layered, genome-resolved analysis. We expanded upon our previous screening of four novel candidate lineages by focusing on the three remaining, uncharacterized environmental isolates, while using the recently validly described Legionella sheltonii (strain PATHC038) as a reference framework control. By applying nucleotide- and protein-based overall genomic relatedness indices (ANI, dDDH, AAI, and POCP) alongside core-genome phylogenomics, our framework independently confirmed the genomic distinctiveness of the three isolates, which exhibited AAI values of 94.3–96.6% and POCP values of 86.5–91.0%, with POCP values lying close to the proposed species delineation threshold. By defining clear criteria for escalating isolates from culture to targeted sequencing, this workflow prevents diagnostic misidentification during epidemiological outbreak investigations and provides public health agencies with a scalable tool to monitor hidden Legionella diversity, ultimately enhancing proactive water safety and risk assessment. Full article

Plant secondary metabolites remain indispensable for pharmaceuticals, nutraceuticals, and cosmeceuticals, yet conventional plant culture systems are increasingly limited by inconsistent yields, poor scalability, and inadequate capacity for real-time process monitoring. Microfluidic technologies and organ-on-a-chip (OoC) platforms, originally developed for mammalian biology, are now emerging as powerful tools to overcome these constraints. These systems enable laminar flow, precise gradient generation, single-cell resolution, and biosensor integration, providing unprecedented control over the cellular microenvironment and supporting non-destructive, real-time metabolomic monitoring. While recent reviews have surveyed plant microfluidics broadly covering developmental biology, single-cell phenotyping, and root–microbe interactions, this review provides, to our knowledge, the first synthesis focused specifically on organ-on-a-chip approaches for plant secondary metabolite biosynthesis and real-time metabolomic monitoring. Advances in device fabrication, including PDMS, paper-based, hydrogel, and thermoplastic materials, surface engineering, gradient-based elicitation strategies, and integration of optical, electrochemical, and mass spectrometric detection systems have also been critically examined. Special emphasis is placed on root-on-a-chip, shoot meristem, protoplast, callus, and 3D organoid platforms for studying cell wall mechanics, vacuolar dynamics, cytoskeletal responses, and signalling cascades. However, challenges remain in long-term culture stability and scalability; nonetheless, these technologies offer a roadmap toward programmable ‘plant biosynthetic factories’ to produce high-value natural products. Full article

Cannabinoid content can vary several-fold across Cannabis sativa L. cultivars that carry functional, closely related CBDA synthase (CBDAS) and THCA synthase (THCAS) alleles, a difference that coding-sequence variation among functional alleles does not fully explain. Structural and copy-number variation at the synthase loci, the functional or pseudogenised state of synthase alleles, and linkage at the B locus account for much of the qualitative drug-versus-hemp chemotype; however, these genetic factors do not fully explain the quantitative, several-fold variation in cannabinoid content among cultivars that carry functional, near-identical synthase alleles. Three independent lines of evidence now indicate that chromatin-level regulation contributes to this variation. H3K4me3 and H3K56ac co-occupy the promoters and gene bodies of THCAS, CBDAS, OLS, and OAC exclusively in glandular trichome tissue while H3K27me3 marks the identical loci in vegetative tissues, indicating that a Polycomb-to-Trithorax chromatin switch is associated with trichome-specific cannabinoid gene expression, potentially independently of transcription factor availability. Progressive DNA hypomethylation accumulates during micropropagation in a cultivar-specific manner, with promoter-region differentially methylated positions reaching up to 22% by twenty subcultures, identifying DNA methylation maintenance as a candidate determinant of epigenetic stability in clonally propagated material, although the functional consequences for cannabinoid pathway gene expression and yield remain to be quantitatively established. In Cannabis indica cell suspension cultures, UV irradiation increases genome-wide DNA methylation (detected by MSAP, which does not resolve locus-specific changes) and elevates CBDAS transcript levels approximately 4-fold relative to non-irradiated controls. This coupling of an environmentally triggered methylation change to cannabinoid pathway gene expression in the Cannabis genus is suggestive, but the genome-wide MSAP signal and the use of C. indica cell cultures mean the locus-specific methylation change at the CBDAS promoter remains to be demonstrated. These findings identify specific, experimentally documented chromatin states as candidate targets for dCas9-effector intervention, including the H3K27me3 repressive state at cannabinoid loci that could be addressed by dCas9-KDM6A and the propagation-induced CG hypomethylation that could be addressed by dCas9-DNMT3A; these editing strategies remain hypotheses that have not yet been tested in Cannabis. We synthesize functional evidence from Catharanthus roseus, Papaver somniferum, and Artemisia annua demonstrating that equivalent chromatin switches control secondary metabolite yield in medicinal plants, evaluate which dCas9-effector architectures are most appropriate for each Cannabis chromatin target, and identify the critical mechanistic gaps that must be closed before epigenome editing can be rationally deployed for cannabinoid yield enhancement in Cannabis. Full article

Floral yeasts are generally recognized to exhibit high metabolic activities. We isolated 437 yeast strains from 45 flower samples collected from different plants in the Beijing Olympic Forest Park, a unique urban ecosystem harboring diverse plants, which facilitated a thorough survey of floral yeast diversity. Based on a sequence analysis of the D1/D2 domain of the large subunit ribosomal RNA gene (LSU rRNA) and the internal transcribed spacer region (ITS), these strains were assigned to 69 species, with Filobasidium magnum, Starmerella bombicola, Teunia globosa, Aureobasidium pini, and Kwoniella ovata being the dominant taxa. Ten representative strains were characterized comprehensively and identified as two novel genera and four novel species. Integrating the molecular data, genome information, and phenotypic/physiological traits, the ten novel yeast strains were described as six novel yeast taxa, including two novel species of two novel genera, Fanglaniella lipolytica gen. nov. sp. nov., Polychromogenomyces tardus gen. nov. sp. nov., and four novel species as Pseudotremella jasmini sp. nov., Teunia pruni sp. nov., Kurtzmanomyces yulaniae sp. nov., and Trigonosporomyces otomorphus sp. nov. Furthermore, qualitative profiling of the lipolytic activity across all six novel taxa was conducted using Tween 20 and Tween 80 plate culture assays, and the lipolytic functional genes were subsequently predicted. Our findings highlighted the high diversity of flower-inhabitant yeast strains and the potential lipolytic activity of floral yeasts. Full article

Three-dimensional scanning, additive manufacturing, and extended reality are now widely used in digital heritage, but they are often discussed as separate technical tools rather than as parts of a user-facing workflow. This review approaches them from the standpoint of user experience, asking how digital capture, model preparation, physical replication, immersive interpretation, and hybrid access affect the way heritage content is understood, used, and trusted. The paper develops a critical narrative discussion of recent work and design practices, without adopting a systematic review protocol. Particular attention is given to educational use, museum interpretation, accessibility, tactile interaction, XR navigation, perceived authenticity, and the evaluation of user experience. The discussion indicates that UX is shaped before the final interface appears: incomplete capture, opaque model editing, poorly readable replicas, or confusing XR layers can all weaken the cultural value of an otherwise advanced system. Recurring barriers include technical complexity, interoperability problems, limited staff training, institutional constraints, accessibility gaps, and uncertainty around automated reconstruction. The review argues that automated digital heritage workflows should be planned as human-centered systems, where efficiency is balanced with usability, inclusion, transparent interpretation, and user-based evaluation. Full article

Background: Dermatophilus congolensis, a Gram-positive actinomycete, is a well-known animal pathogen but a rare cause of human skin infections. Human dermatophilosis is an infection associated with tropical environments and animal contact that remains frequently underdiagnosed. Case Presentation: We report a case of recurrent folliculitis in a 34-year-old male Muay Thai fighter returning from Thailand to Spain. The patient presented with pustular lesions on his lower limbs following frequent leg shaving and close physical contact during training. Microbiological culture from skin swabs yielded beta-hemolytic colonies, identified as D. congolensis by MALDI-TOF MS. Identification was confirmed through Whole-Genome Sequencing (WGS), which also revealed an absence of antimicrobial resistance genes. Despite the lack of established EUCAST breakpoints, the isolate showed low MICs for most tested antibiotics. The patient was treated with doxycycline, although clinical follow-up was not possible due to travel. Conclusions: This case highlights D. congolensis as an emerging differential diagnosis for persistent folliculitis in travelers and athletes. Our findings suggest a potential horizontal transmission route through contact sports and fomites (e.g., mats) in high-humidity settings. Full article

Cumulative reports of psychiatric and neurological outcomes due to synthetic cathinones continue to raise public concern. However, the understanding of the neurotoxic mechanism of action is still poorly understood, particularly for the under-explored αPHP, one of the main MDPV derivatives. In particular, the effects of this synthetic drug on neural stem/progenitor cell cultures are still unknown. Therefore, in the proposed in vitro study, the effects of increasing αPHP concentrations (50–2000 μM) on cell morphology, neuronal/glial differentiation, cell death pathways, and ultrastructure have been evaluated after exposure in murine 2D NSPCs and 3D neurospheres using complementary techniques, i.e., phase contrast microscopy, immunocytochemistry, confocal microscopy, and transmission electron microscopy. We observed that αPHP was able to induce a dose-dependent neurotoxic and neuromodulatory effect in murine 2D NSPC cultures and a 3D neurosphere model, affecting neuronal/glial differentiation, activating the apoptotic pathway, and inducing morphological and ultrastructural changes. The present study could pave the way for a broadened knowledge of synthetic cathinone (SCs) toxicology, needed to establish the right treatment for novel psychoactive substance (NPS) exposure and the possible consequences for public health. Full article

The purpose of this study is to examine the importance of architectural details and artefacts as cultural markers within the interior spaces of adaptively reused heritage buildings in Sofia’s historic urban centre. A comparative study was performed on three significant early twentieth-century residential buildings: the house linked to the former ‘Excelsior’/‘Asen Zlatarov’ cinema complex at Blvd ‘Christo Botev’ N85, the former residence at Blvd ‘Christo Botev’ N75, and the building at Blvd ‘Stamboliyski’ N36—the latter two have both been transformed into hotels. The research employs an interdisciplinary methodology combining archival research, field surveys, photographic documentation, 3D laser scanning, and comparative stylistic analysis. The study’s principal contribution is the development of a transferable analytical framework for classifying interior details by function (constructive or decorative), location (enclosing surfaces, fixed furnishings, vertical circulation), and material—a framework not previously applied to this building stock in the Bulgarian context. The findings confirm that authentic interior details—including stair railings, flooring patterns, gypsum cornices, ornamental plasterwork, heating appliances, and joinery—function as reliable cultural artefacts and markers of historical period and stylistic identity. These details serve as evidence for dating, authenticating, and analysing the evolution of architectural style elements. Preserving and integrating these elements into adapted interiors strengthens architectural identity and supports the sustainable reuse of cultural heritage for contemporary purposes. Full article

Colorectal cancer (CRC) is a major cause of cancer-related mortality among adults younger than 50 years of age, and many tumors show incomplete response or develop resistance to 5-fluorouracil (5-FU)-based chemotherapy. Therefore, new therapeutic approaches that improve CRC sensitivity to existing chemotherapeutic agents are needed. KRAS-associated signaling contributes to CRC growth, metabolic adaptation and treatment resistance. In this study, we investigated whether sulfasalazine (SSZ), a U.S. Food and Drug Administration (FDA)-approved anti-inflammatory drug, could enhance the response of CRC cells to 5-FU and modulate KRAS/mitogen-activated protein kinase (MAPK)-associated signaling. Public dataset analysis using cBioPortal, Kaplan–Meier Plotter and DepMap showed that KRAS is commonly altered in CRC. The analysis also showed that higher KRAS expression was associated with shorter overall survival in 1061 CRC patients, while CRC cell-line models demonstrated KRAS dependency. In 2D cultures, both KRAS-mutant HCT116 and KRAS-wild-type RKO cells showed lower cell viability, reduced colony formation and decreased KRAS expression after SSZ treatment. In 3D cultures, exposure to SSZ reduced early spheroid formation, both as a single treatment and when combined with 5-FU. In established spheroids, SSZ-containing treatments affected cell viability, spheroid growth and morphology, with the most noticeable suppressive effect observed in RKO aggregates. SynergyFinder+ dose-matrix analysis identified dose ranges where SSZ and 5-FU showed additive-to-synergistic effects, leading us to select 600 μM SSZ with 25 μM 5-FU for further validation. Western blot results from PEGDA microwell-derived 3D spheroids showed that SSZ + 5-FU treatment reduced KRAS expression and affected KRAS/MAPK-related signaling. This effect was more pronounced in RKO cells, where downstream pathway suppression was stronger. The combination treatment also increased apoptosis-associated PARP cleavage. At the same time, it reduced Cyclin D1 and GPX4 protein levels and changed the expression of stemness-related markers, including ALDH1A3, CD44, and CD133. Together, these results support SSZ as a candidate repurposed adjuvant that may improve the response to 5-FU in CRC spheroid models and support the use of PEGDA microwell-based 3D platforms for testing combination therapy approaches. Full article

Toll-like receptors (TLRs) are essential in the innate immune response. Furthermore, neuronal TLRs have been involved in regulating neuronal dendritic outgrowth and excitability. However, a deeper understanding of neuronal-TLR function is essential. We investigated the expression and role of TLR8 using mouse postnatal hippocampal neuronal cultures. We assessed TLR8 (and TLR7) expression by immunocytochemistry and TLR8 functionality via electrophysiological recordings in a high-density multi-electrode array system and calcium flux imaging. Pathway activation following fast TLR8 stimulation was performed via Western blot. TLR8 and TLR7 are expressed in postnatal hippocampal neurons at DIV14 and in neurosphere-derived neurons. TLR8 expression was detected in the cell body and dendrites of GABAergic, parvalbumin-expressing interneurons. Stimulation with the TLR8 agonist TL8-506 led to increased neuronal activity in the short term and enhanced neuronal synchronization in short- and long-term recordings. The calcium flux induced by TLR8 was reduced by blocking NMDA receptors with D-AP5. TLR8 stimulation did not lead to IkB-α degradation. Our findings demonstrate that TLR8 and TLR7 are expressed in mouse postnatal hippocampal neurons. TLR8 activation leads to increased neuronal excitability that was decreased by NMDA antagonism. However, acute TLR8 stimulation failed to robustly activate the canonical TLR signaling pathway. We propose that TLR8 activation might lead to the enhancement of excitatory NMDA receptors. Full article

Mobile applications are increasingly promoted as instruments for improving agricultural information access, advisory delivery, market participation, and decision support, particularly for smallholder farmers in developing nations. Research, however, has been dominated by general technology-acceptance and diffusion constructs, while the design-sensitive and infrastructural mechanisms studied in human–computer interaction (HCI) have received comparatively little attention. In this paper we develop a parsimonious HCI model of mobile application use in agriculture. Drawing on the technology acceptance model, the unified theory of acceptance and use of technology, diffusion of innovations, socio-technical systems theory, and human–computer interaction for development (HCI4D), the model proposes that agricultural application use is driven by five antecedent domains: perceived agronomic value, inclusive usability and accessibility, contextual and cultural fit, trust and transparency, and social and institutional embeddedness. Each plays a distinct role across three use stages: adoption intention, sustained use, and decision impact. Contextual constraints (infrastructure and farmer characteristics) moderate these relationships. We develop six testable propositions from the model. The model is conceptual: it is offered as a framework for empirical testing rather than as a validated account of farmer behavior. The paper contributes an HCI-sensitive specification of mobile application use under agricultural field conditions: a “field-ready” conception of mobile HCI. Full article