Search Results (4,765)

Primary ciliary dyskinesia (PCD) is a genetically heterogeneous disorder characterized by impaired mucociliary clearance due to defects in motile cilia. This study investigates the impact of loss-of-function mutations in the CFAP300 gene on the ciliary structure and function in three PCD patients. Using a multimodal approach, we integrated molecular genetic testing, transmission electron microscopy, the high-speed video microscopy assay and immunofluorescence staining to analyze ciliary motility and protein expression in both ex vivo and in vitro-obtained ciliary cells. Our results revealed that the pathogenic variant c.198_200delinsCC (p.Phe67ProfsTer10) in CFAP300 led to the absence of the functional CFAP300 protein, the complete loss of outer and inner dynein arms and immotile cilia. Air–liquid interface (ALI)-cultured cells from patients exhibited no ciliary beating, contrasting with healthy controls. Immunostaining confirmed the absence of CFAP300 in patient-derived cilia, underscoring its critical role in dynein arm assembly. These findings highlight the diagnostic utility of ALI cultures combined with functional and protein analyses for PCD, offering a clinically actionable framework that can be readily incorporated into standard diagnostic workflows. Full article

The retina is highly sensitive to oxygen and blood supply, and hypoxia plays a key role in retinal diseases such as diabetic retinopathy (DR) and age-related macular degeneration (AMD). Müller glial cells, which are essential for retinal homeostasis, respond to injury and hypoxia with reactive gliosis, characterized by the upregulation of the glial fibrillary acidic protein (GFAP) and vimentin, cellular hypertrophy, and extracellular matrix changes, which can impair retinal function and repair. The retinal pigment epithelium (RPE) supports photoreceptors, forms part of the blood–retinal barrier, and protects against oxidative stress; its dysfunction contributes to retinal degenerative diseases such as AMD, retinitis pigmentosa (RP), and Stargardt disease (SD). Extracellular vesicles (EVs) play a crucial role in intercellular communication, protein homeostasis, and immune modulation, and have emerged as promising diagnostic and therapeutic tools. Understanding the role of extracellular vesicles’ (EVs’) signaling machinery of glial cells and the retinal pigment epithelium (RPE) is critical for developing effective treatments for retinal degeneration. In this study, we investigated the bidirectional EV-mediated crosstalk between RPE and Müller cells under hypoxic conditions and its impact on cellular metabolism and retinal cell integrity. Our findings demonstrate that RPE-derived extracellular vesicles (RPE EVs) induce time-dependent metabolic reprogramming in Müller cells. Short-term exposure (24 h) promotes pathways supporting neurotransmitter cycling, calcium and mineral absorption, and glutamate metabolism, while prolonged exposure (72 h) shifts Müller cell metabolism toward enhanced mitochondrial function and ATP production. Conversely, Müller cell-derived EVs under hypoxia influenced RPE metabolic pathways, enhancing fatty acid metabolism, intracellular vesicular trafficking, and the biosynthesis of mitochondrial co-factors such as ubiquinone. Proteomic analysis revealed significant modulation of key regulatory proteins. In Müller cells, hypoxic RPE-EV exposure led to reduced expression of Dyskerin Pseudouridine Synthase 1 (DKc1), Eukaryotic Translation Termination Factor 1 (ETF1), and Protein Ser/Thr phosphatases (PPP2R1B), suggesting alterations in RNA processing, translational fidelity, and signaling. RPE cells exposed to hypoxic Müller cell EVs exhibited elevated Ribosome-binding protein 1 (RRBP1), RAC1/2, and Guanine Nucleotide-Binding Protein G(i) Subunit Alpha-1 (GNAI1), supporting enhanced endoplasmic reticulum (ER) function and cytoskeletal remodeling. Functional assays also revealed the compromised barrier integrity of the outer blood–retinal barrier (oBRB) under hypoxic co-culture conditions. These results underscore the adaptive but time-sensitive nature of retinal cell communication via EVs in response to hypoxia. Targeting this crosstalk may offer novel therapeutic strategies to preserve retinal structure and function in ischemic retinopathies. Full article

Chronic infection with the hepatitis B virus (HBV) results in over 1 million deaths annually. Although currently licensed treatments, including pegylated interferon-α and nucleoside/nucleotide analogs, can inhibit viral replication, they rarely eradicate covalently closed circular DNA (cccDNA) reservoirs. Moreover, vaccination does not offer therapeutic benefit to already infected individuals or non-responders. Consequently, chronic infection is maintained by the persistence of cccDNA in infected hepatocytes. For this reason, novel therapeutic strategies that permanently inactivate cccDNA are a priority. Obligate heterodimeric transcription activator-like effector nucleases (TALENs) provide the precise gene-editing needed to disable cccDNA. To develop this strategy using a therapeutically relevant approach, TALEN-encoding mRNA targeting viral core and surface genes was synthesized using in vitro transcription with co-transcriptional capping. TALENs reduced hepatitis B surface antigen (HBsAg) by 80% in a liver-derived mammalian cell culture model of infection. In a stringent HBV transgenic murine model, a single dose of hepatotropic lipid nanoparticle-encapsulated TALEN mRNA lowered HBsAg by 63% and reduced viral particle equivalents by more than 99%, without evidence of toxicity. A surveyor assay demonstrated mean in vivo HBV DNA mutation rates of approximately 16% and 15% for Core and Surface TALENs, respectively. This study presents the first evidence of the therapeutic potential of TALEN-encoding mRNA to inactivate HBV replication permanently. Full article

A just and sustainable energy transition in historically coal-dependent regions like Kentucky requires more than the adoption of new technologies and market-based solutions. This study uses a stated preferences approach to evaluate public support for various attributes of energy transition programs, revealing broad backing for moving away from coal, as indicated by a negative willingness to pay (WTP) for the status quo (–USD 4.63). Key findings show strong bipartisan support for solar energy, with Democrats showing the highest WTP at USD 8.29, followed closely by Independents/Others at USD 8.22, and Republicans at USD 8.08. Wind energy also garnered support, particularly among Republicans (USD 4.04), who may view it as more industry-compatible and less ideologically polarizing. Job creation was a dominant priority across political affiliations, especially for Independents (USD 9.07), indicating a preference for tangible, near-term economic benefits. Similarly, preserving cultural values tied to coal received support among Independents/Others (USD 4.98), emphasizing the importance of place-based identity in shaping preferences. In contrast, social support programs (e.g., job retraining) and certain post-mining land uses (e.g., recreation and conservation) were less favored, possibly due to their abstract nature, delayed benefits, and political framing. Findings from Kentucky offer insights for other coal-reliant states like Wyoming, West Virginia, Pennsylvania, Indiana, and Illinois. Ultimately, equitable transitions must integrate local voices, address cultural and economic realities, and ensure community-driven planning and investment. Full article

This study aims to understand the meaning nursing professionals attribute to their lived experiences of providing humanized care within a public hospital setting. Grounded in Jean Watson’s theory of human caring, the research adopts a qualitative, descriptive phenomenological design to capture the perceptions and emotions of nurses regarding humanized care. Data were collected through semi-structured interviews with nine experienced nurses, selected through purposive sampling. The interviews, conducted virtually between July and December 2024, were analyzed using Colaizzi’s method and supported by Atlas.ti software. Four main thematic categories emerged: institutional health policies, professional image and identity, strengths and challenges in care, and essential competencies for humanized care. The findings highlight the critical role of empathy, cultural sensitivity, ethical commitment, and emotional presence in delivering compassionate care. Participants emphasized that, beyond clinical procedures, humanized care requires relational and contextual sensitivity, often hindered by institutional limitations and excessive administrative burdens. The study concludes that nursing professionals are key agents in promoting ethical, empathetic, and culturally respectful practices that humanize health services. These insights offer valuable contributions for designing policies and training strategies aimed at strengthening humanized care as a cornerstone of quality healthcare systems. Full article

Globally, technologically integrated housing solutions are increasingly relevant in addressing the challenges of aging populations and sustainable urban development. Drawing on the Technology Acceptance Model (TAM), this research investigates how perceptions of usefulness, ease of use, and attitudes influence relocation intention to smart retirement villages (SRVs), while also examining any significant differences between the socio-demographic variables and such intention. A total of 305 individuals aged 55 and above participated in an online survey, with data analyzed using IBM SPSS Statistics version 27 and AMOS-SEM version 25. The findings reveal that elderly individuals of Chinese ethnicity, those who are married, and those aged between 66 and 70 are more inclined to relocate to SRVs. Attitude and perceived usefulness significantly predict relocation intention, while perceived ease of use exerts an indirect effect through usefulness. These results highlight the importance of integrating user-centered technological design with socio-cultural and demographic considerations in the development of age-friendly built environments. The study offers insights for urban planners, policymakers, and developers seeking to create inclusive and sustainable smart housing solutions for aging populations. Full article

Coupled grey and green infrastructure (CGGI) offers a promising pathway toward sustainable stormwater management in historic urban environments. This study compares CGGI and conventional grey infrastructure (GREI)-only strategies across four degrees of layout centralization (0%, 33.3%, 66.7%, and 100%) in the Quanzhou West Street Historic Reserve, China. Using a multi-objective optimization framework integrating SWMM simulations, life-cycle cost (LCC) modeling, and resilience metrics, we found that the decentralized CGGI layouts reduced the total LCC by up to 29.6% and required 60.7% less green infrastructure (GI) area than centralized schemes. Under nine extreme rainfall scenarios, the GREI-only systems showed slightly higher technical resilience (Tech-R: max 99.6%) than CGGI (Tech-R: max 99.1%). However, the CGGI systems outperformed GREI in operational resilience (Oper-R), reducing overflow volume by up to 22.6% under 50% network failure. These findings demonstrate that decentralized CGGI provides a more resilient and cost-effective drainage solution, well-suited for heritage districts with spatial and cultural constraints. Full article

Recent advancements in generative artificial intelligence have significantly progressed the field of text-to-music generation, enabling users to create music from natural language descriptions. Despite the success of various models, such as MusicLM, MusicGen, and AudioLDM, the current approaches struggle to capture fine-grained genre-specific characteristics, precisely control musical attributes, and handle underrepresented cultural data. This paper introduces a novel, lightweight fine-tuning method for the AudioLDM framework using low-rank adaptation (LoRA). By updating only selected attention and projection layers, the proposed method enables efficient adaptation to musical genres with limited data and computational cost. The proposed method enhances controllability over key musical parameters such as rhythm, emotion, and timbre. At the same time, it maintains the overall quality of music generation. This paper represents the first application of LoRA in AudioLDM, offering a scalable solution for fine-grained, genre-aware music generation and customization. The experimental results demonstrate that the proposed method improves the semantic alignment and statistical similarity compared with the baseline. The contrastive language–audio pretraining score increased by 0.0498, indicating enhanced text-music consistency. The kernel audio distance score decreased by 0.8349, reflecting improved similarity to real music distributions. The mean opinion score ranged from 3.5 to 3.8, confirming the perceptual quality of the generated music. Full article

As organizations continue to embrace digital transformation, the need for robust cybersecurity strategies has never been more critical. This paper explores the Zero Trust Architecture (ZTA) as a contemporary cybersecurity framework that addresses the challenges posed by increasingly interconnected systems. Zero Trust (ZT) operates under the principle of “never trust, always verify,” ensuring that every access request is thoroughly authenticated, regardless of the requester’s location within or outside the network. However, implementing ZT is a challenging task, requiring an adequate roadmap to prioritize the different initiatives in agreement with company culture, exposure and cyber posture. We apply multi-criteria decision analysis (MCDA) to evaluate the relative importance of various components within a ZT framework, using the Incomplete Analytic Hierarchy Process (IAHP). Expert opinions from professionals in cybersecurity and IT governance were gathered through structured questionnaires, leading to a prioritized ranking of the eight key ZT pillars, as defined by the Cybersecurity and Infrastructure Security Agency (CISA), Washington, DC, USA, along with a prioritization of the sub-elements within each pillar. The study provides actionable insights into the implementation of ZTA, helping organizations prioritize security efforts to mitigate risks effectively and build a resilient digital infrastructure. The evaluation results were used to create a prioritized framework, integrated into the ZEUS platform, developed with Teleconsys S.p.A., to enable detailed assessments of a firm’s cyber partner regarding ZT and identify improvement areas. The paper concludes by offering recommendations for future research and practical guidance for organizations transitioning to a ZT model. Full article

Background: Prebiotics are selectively used by host microorganisms to promote health. Because effective prebiotic doses (1.5–30 g/day) often require inconvenient delivery formats, this study aims to explore whether capsule-compatible doses of galacto-oligosaccharides (GOS) can effectively modulate the gut microbiome. Methods: The impact of Bimuno^® GOS (Reading, UK) at 0.5, 0.75, 1.83, and 3.65 g on the adult gut microbiome was assessed using the ex vivo SIFR^® technology (n = 8), a clinically validated, bioreactor-based technology. Results: The GOS were rapidly fermented and significantly increased beneficial Bifidobacterium species (B. adolescentis, B. bifidum, and B. longum), even at the lowest tested dose. In doing so, GOS strongly promoted SCFA production, particularly acetate (significant from 0.5 g) and butyrate (significant from 0.75 g). Gas production only mildly increased, likely as Bifidobacterium species do not produce gases. Based on the ability of the SIFR^® technology to cultivate strictly anaerobic, hard-to-culture gut microbes, unlike in past in vitro studies, we elucidated that GOS also enriched specific Lachnospiraceae species. Besides Anaerobutyricum hallii, this included Bariatricus comes, Blautia species (B. massiliensis, Blautia_A, B. faecis), Oliverpabstia intestinalis, Mediterraneibacter faecis, and Fusicatenibacter species. Finally, GOS also promoted propionate (significant from 0.75 g), linked to increases in Phocaeicola vulgatus. Conclusions: GOS displayed prebiotic potential at capsule-compatible doses, offering greater flexibility in nutritional product formulation and consumer convenience. Notably, the strong response at the lowest dose suggests effective microbiome modulation at lower levels than previously expected. Full article

Cancer is a major global health issue, with rising incidence rates highlighting the urgent need for more effective treatments. Despite advances in cancer therapy, challenges such as adverse effects and limitations of existing treatments remain. Immunotherapy, which harnesses the body’s immune system to target cancer cells, offers promising solutions. Gamma delta (γδ) T cells are noteworthy due to their potent ability to kill various cancer cells without needing conventional antigen presentation. Recent studies have focused on the role of γδ T cells in α-galactosylceramide (α-GalCer)-mediated immunity, opening new possibilities for cancer immunotherapy. We engineered humanized T cell receptor (HuTCR)-T1 γδ mice by replacing mouse sequences with human counterparts. This study investigates the cytotoxic activity of humanized γδ T cells against several human cancer cell lines (A431, HT-29, K562, and Daudi) in vitro, aiming to elucidate mechanisms underlying their anticancer efficacy. Human cancer cells were co-cultured with humanized γδ T cells, with and without α-GalCer, for 24 h. The humanized γδ T cells showed enhanced cytotoxicity across all tested cancer cell lines compared to wild-type γδ T cells. Additionally, γδ T cells from HuTCR-T1 mice exhibited higher levels of anticancer cytokines (IFN-γ, TNF-α, and IL-17) and Granzyme B, indicating their potential as potent mediators of anticancer immune responses. Blocking γδ T cells’ cytotoxicity confirmed their γδ-mediated function. These findings represent a significant step in preclinical development of γδ T cell-based cancer immunotherapies, providing insights into their mechanisms of action, optimization of therapeutic strategies, and identification of predictive biomarkers for clinical application. Full article

Human cutaneous leishmaniasis (CL) caused by Leishmania (Viannia) braziliensis is a complex parasitic disease marked by dynamic host–parasite interactions and immunomodulation. Extracellular vesicles (EV) derived from immune cells have emerged as key mediators of intercellular communication and potential biomarkers in infectious diseases. In this study, we combined a modified lymphocyte proliferation assay with nano-flow cytometry to quantify and phenotype EV released by CD4⁺, CD8⁺, and CD14⁺ cells in PBMC cultures from CL patients at different clinical stages: before treatment (PBT), during treatment (PDT), and post-treatment (PET) with antimonial. Healthy individuals (HI) were included as physiological controls. Upon stimulation with L. (V.) braziliensis antigens, we observed a distinct modulation of EV subsets. In the PBT group, CD4⁺ and CD14⁺ EV were significantly reduced, while CD8⁺ EV remained elevated. During PDT and PET, EV concentrations were restored across all subsets. These findings suggest that L. (V.) braziliensis selectively modulates the release of immune cell–derived EV, possibly as an immune evasion mechanism. The restoration of EV release following antimonial therapy highlights their potential as sensitive biomarkers for disease activity and treatment monitoring. This study offers novel insights into the immunoregulatory roles of EV in CL and underscores their relevance in host–parasite interactions. Full article

This paper investigates the cultural, spiritual, and ecological use and value of fish by-products in the material practices of Alaska Native (Indigenous Peoples are the descendants of the populations who inhabited a geographical region at the time of colonisation and who retain some or all of their own social, economic, cultural, and political institutions. In this paper, I use the terms “Indigenous” and “Native” interchangeably. In some countries, one of these terms may be favoured over the other.) and Greenlandic Inuit women. It aims to uncover how fish remnants—skins, bones, bladders, vertebrae, and otoliths—were transformed through tanning, dyeing, and sewing into garments, containers, tools, oils, glues, and adornments, reflecting sustainable systems of knowledge production rooted in Arctic Indigenous lifeways. Drawing on interdisciplinary methods combining Indigenist research, ethnographic records, and sustainability studies, the research contextualises these practices within broader environmental, spiritual, and social frameworks. The findings demonstrate that fish-based technologies were not merely utilitarian but also carried symbolic meanings, linking wearers to ancestral spirits, animal kin, and the marine environment. These traditions persisted even after European contact and the introduction of glass trade beads, reflecting continuity and cultural adaptability. The paper contributes to academic discourse on Indigenous innovation and environmental humanities by offering a culturally grounded model of zero-waste practice and reciprocal ecology. It argues that such ancestral technologies are directly relevant to contemporary sustainability debates in fashion and material design. By documenting these underexamined histories, the study provides valuable insight into Indigenous resilience and offers a critical framework for integrating Indigenous knowledge systems into current sustainability practices. Full article

Background/Objectives: Senior nurses in Taiwan shoulder layered responsibilities shaped by professional roles, gendered expectations, and family duty. Although Taiwan faces a persistent shortage of experienced clinical nurses, limited research has explored how long-serving nurses sustain identity and commitment across decades of caregiving. This study examines how senior staff nurses understand their journeys of becoming—and remaining—nurses within a culturally and emotionally complex landscape. Methods: Interviews were conducted between May 2019 and September 2023 in locations chosen by participants, with most sessions face-to-face and others undertaken via video conferencing during COVID-19. This narrative inquiry involved in-depth, multi-session interviews with five female senior staff nurses born in the 1970s to early 1980s. Each participant reflected on her life and career, supported by co-constructed “nursing life lines.” Thematic narrative analysis was conducted using McCormack’s five-lens framework and Riessman’s model, with ethical rigor ensured through reflexive journaling and participant validation. Results: Three overarching themes emerged: (1) inner strength and endurance, highlighting silent resilience and the ethical weight of caregiving; (2) support and responsibility in relationships, revealing the influence of family, faith, and relational duty; and (3) role navigation and professional identity, showing how nurses revisit meaning, self-understanding, and tensions across time. Participants described emotionally powerful moments, identity re-connection, and cultural values that shaped their paths. Conclusions: These narratives offer a relational and culturally embedded understanding of what it means to sustain a career in nursing. Narrative inquiry created space for reflection, meaning-making, and voice in a system where such voices are often unheard. Identity was not static—it was lived, reshaped, and held in story. Full article

Urban forests, as a form of green infrastructure, play a vital role in enhancing urban resilience, environmental health, and quality of life. However, users perceive and utilize these spaces in diverse ways. This study aims to identify latent perception types among urban forest visitors and analyze their behavioral, demographic, and policy-related characteristics in Incheon Metropolitan City (Republic of Korea). Using latent class analysis, four distinct visitor types were identified: multipurpose recreationists, balanced relaxation seekers, casual forest users, and passive forest visitors. Multipurpose recreationists preferred active physical use and sports facilities, while balanced relaxation seekers emphasized emotional well-being and cultural experiences. Casual users engaged lightly with forest settings, and passive forest visitors exhibited minimal recreational interest. Satisfaction with forest elements such as vegetation, facilities, and management conditions varied across visitor types and age groups, especially among older adults. These findings highlight the need for perception-based green infrastructure planning. Policy recommendations include expanding accessible neighborhood green spaces for aging populations, promoting community-oriented events, and offering participatory forest programs for youth engagement. By integrating user segmentation into urban forest planning and governance, this study contributes to more inclusive, adaptive, and sustainable management of urban green infrastructure. Full article