Search Results (310)

Conventional anaerobic digestion relies on diverse inocula present in sludge-based systems. A defined consortium approach was investigated as an alternative. Acetivibrio cellulolyticus was chosen as the cellulose degrader, and two strains of Methanosarcina barkeri were selected as methane producers. Initial cultivation following manufacturer protocols highlighted significant challenges in maintaining strict anaerobic conditions, particularly in the absence of specialized infrastructure. A simplified anaerobic cultivation workflow was therefore evaluated for pure cultures of the selected anaerobes and subsequently used to evaluate a defined consortium using microcrystalline cellulose (MCC) and industrial lignocellulosic biomass (LB) residue as growth substrates. The workflow enabled successful cultivation of pure cultures in their recommended nutrient media without detectable contamination, as assessed by microscopy, aerobic contamination checks, and gas chromatography analysis. Growth-associated observations were obtained on MCC after prolonged incubation (~30 days); however, a metabolically active cellulolytic–methanogenic consortium was not established, as methane was not detected and no activity was detected on the LB substrate. This study demonstrates that anaerobic cultivation of fastidious microorganisms is feasible using a simplified method without fully controlled anaerobic environments and highlights inherent challenges associated with the defined consortium on substrates such as MCC and complex LB residue. The simplified workflow may provide an accessible approach to anaerobic cultivation for sustainable biomethane research in laboratories lacking specialized anaerobic infrastructure. Further work is required to determine conditions supporting methane production using the defined consortium. Full article

Landscape Character Assessment (LCA) has struggled with scale mismatches and inadequate treatment of cultural expression when applied to high-density cultural landscapes in Asia. On the Chengdu Plain, Western Sichuan, China, remote sensing shows convergent settlement patterns across case sites, but fieldwork reveals substantial divergences in landscape character. We examined five cases representing different rural development models in the Chengdu metropolitan area. By integrating 10 m Sentinel-2 imagery and with systematic fieldwork (3200 photographs and 42 semi-structured interviews, July–August 2023), we constructed a framework that combines macro-scale spatial analysis with micro-scale field investigation. For macro-level quantification, six landscape pattern indices were adopted: number of patches (NP), patch density (PD), contagion index (CONTAG), aggregation index (AI), Shannon’s diversity index (SHDI), and Shannon’s evenness index (SHEI). The micro-level investigation followed a three-tier scheme (natural environment, settlement space, architectural details) to record vertical ecological communities, spatial expressions of cultural embeddedness, and visual landscape character. Indicator screening employed the KJ method and two rounds of Delphi consultation with ten experts; weights were then determined through the Analytic Hierarchy Process (AHP). Macro-level results display a clustered, low-fragmentation pattern (woodland AI = 97.48, water body AI = 99.40), anchored by a stable mosaic of Linpan, farmland, water bodies, and homesteads. Micro-level features include multi-strata vertical communities of trees, shrubs, grasses, and water, along with cultural embeddedness expressed through irrigation systems, farming patterns, and courtyard layouts. The VLEAS (Vernacular Landscape Elements Assessment System) framework encompasses three dimensions (natural, cultural, and visual) and comprises 12 indicators. The cultural dimension receives the highest weight (0.42), followed by natural (0.32) and visual (0.26) dimensions. Comparative analysis shows that the framework differentiates cases with similar macro-scale patterns but divergent micro-features, capturing differences that a purely macro-index benchmark misses, and it also reveals trade-offs among ecological integrity, cultural heritage, and visual quality. Sensitivity analysis displays that case rankings remain stable across four extreme weighting scenarios (cultural, ecological, visual, equal), with only one swap (Xingfu Pastoral Park and Nongke Village) under ecological priority and no rank change greater than one position, confirming that the results are not driven by any specific weight assignment. This approach extends the utility of LCA in high-density rural cultural landscapes and offers a practical tool for differentiated conservation and management, especially in tourism-influenced rural settings on the Chengdu Plain. Transferability to other regions would require recalibration of locally relevant indicators. Full article

Burkholderia gladioli pv. cocovenenans (B. gladioli pv. cocovenenans), the most representative pathovar of B. gladioli, is capable of producing thermostable and highly virulent bongkrekic acid, which can result in multiple organ failure and an extremely high case fatality rate in humans. This research aims to mine specific targets for B. gladioli pv. cocovenenans and develop a rapid and accurate assay for the detection of B. gladioli pv. cocovenenans in food. Two novel specific target genes (group_0918 and group_0110) were successfully screened through pan-genome analysis, and the excellent specificity and sensitivity of both targets were verified in comparison to PCR assays. Based on these targets, a loop-mediated isothermal amplification (LAMP) method was developed to specifically identify B. gladioli pv. cocovenenans. In pure culture and spiked fresh black fungus samples, the LAMP assay achieved a limit of detection (LOD) of 4.1 × 10¹ CFU/mL and 4.1 × 10² CFU/g for the detection of B. gladioli pv. cocovenenans, respectively. To enable point of care testing, a LAMP-based lateral flow dipstick(LFD) assay was established with a detection sensitivity of 4.1 × 10³ CFU/g in fresh black fungus. Tests on naturally infected food samples demonstrated that the developed LAMP and LAMP-assisted visual detection assays can serve as an effective alternative for the accurate identification of B. gladioli pv. cocovenenans. The novel molecular detection technology established in this study offers an efficient and reliable detection method for the prevention and control of B. gladioli pv. cocovenenans contamination in food, which holds significant application value and promotion prospects. Full article

Background/Objectives: This study aimed to evaluate the direct effect of elements released from dental casting alloys on human gingival fibroblasts (HGF). Methods: Ten discs each of commercially pure titanium (Ti), silver-palladium (Ag-Pd), and nickel-chromium (Ni-Cr) alloys, measuring 4 mm in diameter and 3 mm in thickness, were fabricated and placed directly on HGF cultures for 72 h. Five discs from each alloy group were incubated in DMEM, whereas the remaining five discs were incubated in artificial saliva. Cytotoxicity was assessed using three in vitro methods: morphological evaluation of HGF, the dye exclusion test (DET), and the methyl-thiazolyl-tetrazolium (MTT) colorimetric assay. Results: HGF directly exposed to cpTi, Ag-Pd, and Ni-Cr discs demonstrated reduced cell density around the alloy specimens compared with untreated control cells after 72 h of incubation in both DMEM and artificial saliva. cpTi alloy exhibited no detectable cytotoxic effects, whereas Ag-Pd and Ni-Cr alloys demonstrated cytotoxicity in both DMEM and artificial saliva under the experimental conditions. Among the tested alloys, Ni-Cr exhibited the highest cytotoxicity. Artificial saliva enhanced the cytotoxic effects of the tested alloys, as evidenced by increased morphological alterations, reduced cell viability, and lower MTT activity compared with DMEM. Conclusions: cpTi alloy exhibited the lowest direct cytotoxic effect on HGF in both DMEM and artificial saliva. In contrast, Ag-Pd and Ni-Cr alloys demonstrated greater cytotoxicity, particularly in the presence of artificial saliva, suggesting that both alloy composition and the surrounding environment influence the biological response of oral soft tissues. Full article

Regenerating fragile historical contexts requires choices of repurposing that combine heritage protection, continuity of use and managerial feasibility, in the presence of multiple objectives and stakeholders with different preferences. This study develops and tests an MCDA-based decision-support framework for the ex-ante selection of adaptive reuse scenario applied to Craco (Italy) and Palazzo Carbone-Rigirone. Craco and Palazzo Carbone-Rigirone were selected as a critical case because they combine heritage abandonment, geomorphological fragility, cultural visibility, weak local services and the need for a feasible management model. The methodology involves: (i) defining four adaptive reuse scenarios; (ii) constructing nine criteria that integrate socio-economic impacts, safety/security, cultural attractiveness, compatibility with the property and economic–financial feasibility; (iii) elicitation of weights using a hybrid approach, combining the decision-maker’s macro priorities and the social quota derived from questionnaires using normalised indicators of satisfaction/dissatisfaction and priorities for improvement; (iv) classification using the Weighted Sum Model and TOPSIS under two normalizations (distributive and ideal) and two variants (relative and absolute). The results show convergence between methods and stability of the ranking, with a preference for the multifunctional scenario oriented towards cultural services and socialising. In the case of Craco, adaptive reuse offers advantages compared with purely conservative, passive musealization or tourism-only strategies. The study concludes that MCDA is useful as a transparent pre-selection tool and supports the alignment of local needs and institutional priorities; its robustness can be strengthened with sensitivity analyses and policy scenarios. Full article

Background/Objectives: Digital technologies and data-driven approaches are rapidly transforming healthcare practice and enabling more personalized and preventive care. As personal health data becomes increasingly embedded in healthcare systems, understanding how future healthcare professionals interpret these developments is essential for shaping responsive health education. This study aims to identify the factors that students in health-related programs consider important regarding the increasing use of personal health data in healthcare. Methods: An exploratory qualitative focus group study was conducted between March 2024 and July 2025 across five higher education institutions in Australia, Canada, and the Netherlands. Seven focus groups were conducted with forty students from health-related programs, including nursing, public health, occupational therapy, and social work. Participants discussed the use of personal health data in healthcare and reflected on short fictional future scenarios designed to stimulate discussion about possible developments in data-driven healthcare. Data were analyzed using reflexive thematic analysis using ATLAS.ti. Results: Three overarching domains were identified: (1) personalization and prevention, (2) data quality and ethical considerations, and (3) organizational implications and conditions. Students described personal health data as a powerful tool for personalization, prevention, and informed decision-making. At the same time, they raised concerns about data reliability, overreliance on automated systems, patient anxiety, potential dehumanization of care, privacy risks, and emerging inequalities related to access to and representation within data systems. Overall, students appeared neither purely techno-optimistic nor technophobic, but articulated nuanced ethical, cultural, and professional tensions surrounding data-driven care. Conclusions: Preparing future healthcare professionals for data-driven healthcare requires integrating critical data literacy, ethical reflection, interdisciplinary collaboration and opportunities to critically engage with the societal and professional implications of data-driven technologies into health professional education, while ensuring that organizational conditions support the responsible use of personal health data. Full article

This paper presents a multidimensional analysis of Historic Building Information Modeling (HBIM) segmentation, offering a roadmap towards standardization, a key dimension towards broader adoption within the Cultural Heritage (CH) sector. HBIM faces multiple challenges related to the lack of standardized protocols and varying definitions of Level of Detail (LOD) across applications. Amid the advancements of the fourth industrial revolution, integrating Building Information Modeling (BIM) improves sustainability and digital governance, aligning with the sustainable development agenda. Despite increasing academic interest, the implementation of HBIM remains limited, primarily due to the complexities and heterogeneities inherent in CH artifacts. This study begins with a purely qualitative strategy. Then, it introduces multidimensional and hierarchical clustering analysis to classify the unique characteristics of various HBIM applications such as segmentation, input, and data-capturing media. At the same time, it is a tool for fine-tuning keyword-based selection criteria, which is crucial in systematic or semi-systematic surveys in HBIM segmentation. The thematic analysis output is interrupted just before the conceptualization step, and theme extraction is diverted to correspondence analysis implemented in R, an open-source statistical package. Among the key findings of this paper is the classification of four distinct HBIM application clusters, revealing how specific workflows align with data acquisition methods, input formats, and Level of Detail (LOD) requirements. The analysis exposes critical standardization bottlenecks hindering wider-scale industry adoption, highlighting that challenges are domain-specific. Strong evidence shows that 3D modeling has not reached the required maturity level, with persisting challenges distributed non-uniformly within the applications spectrum. Finally, AI-driven automation relates with poor LOD outcome. Full article

With the growing interest in the microbiome, short-chain fatty acids (SCFAs) have emerged as key metabolites due to their critical roles in host physiology, including immune regulation, energy homeostasis, and inflammatory control. As a result, the accurate quantification of SCFAs in various biological samples has become increasingly important. However, reliable and standardized methods for measuring SCFAs across different sample types remain underdeveloped, highlighting the need for methodological refinement. To address this need, we optimized two analytical methods, headspace GC-MS and GC-MS/MS, for SCFA quantification. These techniques were applied to a range of biological matrices, including pure microbial cultures, low-abundance animal liver, animal feces, and standardized simulated human fecal samples. The headspace GC-MS approach enables direct analysis with minimal sample preparation, thereby enhancing throughput and ease of use. In contrast, the GC-MS/MS method, involving methanol extraction, alkaline treatment, and derivatization with MTBSTFA, offers superior sensitivity and precision, making it particularly suitable for small-volume and low-abundance samples. Together, these optimized protocols provide robust, sensitive platforms for profiling SCFAs across diverse biological matrices, facilitating a deeper understanding of microbiome–host interactions and supporting future translational applications. Full article

Background: Necrotizing fasciitis (NF) is a rapidly progressive, life-threatening soft tissue infection characterized by fascial necrosis, with mortality rates of 20–30%. Despite its rarity, NF is increasingly encountered due to the rising prevalence of predisposing factors. Data from Southern European tertiary centers remain scarce. Methods: We retrospectively reviewed all patients ≥18 years with radiological and/or surgical diagnosis of NF managed at IRCCS Policlinico San Matteo, Pavia, Italy, between November 2018 and August 2023. Clinical, microbiological, and treatment data were extracted from electronic medical records. The Laboratory Risk Indicator for Necrotizing Fasciitis (LRINEC) score was calculated retrospectively. The Charlson Comorbidity Index was computed for each patient. Given the small sample size, we adopted a purely descriptive analytical approach without inferential testing. Results: Thirteen patients met inclusion criteria (median age 58 years, IQR 44.5–79.5; 69.2% male). The most common comorbidities were diabetes mellitus (6/13, 46.2%), renal failure (4/13, 30.8%), and chronic liver disease (4/13, 30.8%). The age-adjusted Charlson Index ranged from 0 to 11 (median 4). Lower limbs were the most frequently affected anatomic site (5/13, 38.5%), followed by the perineal/genital region (Fournier gangrene, 4/13, 30.8%). Type II (monomicrobial) NF predominated (9/13, 69.2%). Microbiological cultures were positive in 8/13 patients (61.5%): Gram-positive cocci were isolated in 5/8 (62.5%) and mixed aerobic/anaerobic flora in 3/8 (37.5%). Empirical antibiotic regimens included a piperacillin–tazobactam backbone in 6/12 (50.0%) patients and a meropenem-based combination in 5/12 (41.7%); 6/12 patients underwent targeted de-escalation after culture results. Two patients (15.4%) died in hospital, both with Fournier gangrene and Type I infection (mortality 2/4, 50.0% in Type I vs. 0/9 in Type II). The median length of stay was 26 days (IQR 17–28.5). All patients had LRINEC ≥6 at admission, with 9/13 (69.2%) classified as high risk (≥8). Conclusions: In this small retrospective Italian cohort, NF was most frequently associated with diabetes and high comorbidity burden. Type I (polymicrobial) infections, predominantly involving the perineal region, showed worse outcomes than Type II infections. The clinical experience accumulated during this study period subsequently informed the development of an institutional empirical antimicrobial protocol for skin and soft tissue infections at our hospital. Full article

The ability to rapidly estimate bacterial numbers in a pure culture is an important research tool, often performed by measuring the optical density of the culture. However, this method is of limited use with mycoplasma cultures. Therefore, a new method for estimating Mycoplasma gallisepticum cell numbers in a pure culture was developed based on the fluorescent measurement of genomic DNA from lysed cells. Actual Mycoplasma gallisepticum counts obtained from either Color Change Units (CCU) or Colony Forming Units (CFU) were used to create equations to estimate mycoplasma concentration from either DNA concentration data obtained from lysed cells or optical density data from mycoplasma in media. The results suggest that calculated counts are slightly more accurate than those obtained from OD600 data. The results further show that calculating culture concentration using the DNA concentration has a wider range compared to using OD600 data. Results also showed that equations generated using one M. gallisepticum strain could work for a second M. gallisepticum strain. However, it was also shown that the equations were not accurate for a different mycoplasma strain. These results suggest that measuring DNA concentration from lysed mycoplasma cells can provide another useful tool for estimating mycoplasma culture concentrations. Full article

Background and Aims: Anemia is seen in nearly >70% of patients with cirrhosis and is often non-responsive to nutritional supplements; therefore, we assessed the erythropoiesis and associated alteration in bone marrow (BM). Methods: It is a cross-sectional study. Flow cytometry was performed to assess the hematopoietic stem cells (HSCs) and erythroid population of 60 patients with cirrhosis compared with patients with 7 non-cirrhotic portal fibrosis (NCPF) and 3 controls. Proteomics were performed of the pure CD71 erythroid population taken from patients with cirrhosis to decipher the internal abnormalities supported by validation experiments. Real Time PCR, colony assay and heme quantification, cytokine array, and ELISA were performed to assess erythropoietic stimulating agents (ESA), inflammatory cytokines, and growth factors as an external factor affecting erythropoiesis. Results: We found a decrease in intermediate erythroid progenitors [IEPs; CD71+ CD235a+], conversely early erythroid precursors [EEP; CD71+ CD235a−] and late erythroid progenitors [LEP; CD71− CD235a+] were increased (p < 0.05) in cirrhotic and NCPF as compared to control. However, unlike NCPF, cirrhosis exhibited decreased CD71+ transferrin receptor (TfR1) expression over erythroid cells and increased immature erythrocytes (p < 0.05) in peripheral circulation. In vitro culture of erythroid precursors showed impaired differentiation and maturation that was confirmed by the reduced (p < 0.05) number of erythroid colonies (BFU-E). Proteomics analysis showed downregulated proteins associated with hemoglobin synthesis, ROS detoxification, translation, and mitochondrial activity. Furthermore, we found an altered expression of genes related to erythropoiesis and hemoglobin synthesis and increase (p < 0.05) in inflammatory cytokines such as IL-5, TRAIL-R2, TGF-α, and TGF-β in BM. Conclusions: This study suggests that the dysregulated erythropoiesis observed in patients with cirrhosis having anemia is maintained despite adequate nutrition. Full article

This study examines the relationship between spatial morphology and cultural sustainability in three heritage Nubian villages—Gharb Seheyl, Elephantine Island, and Heisa Island. Addressing the limitations of purely quantitative or qualitative approaches, it establishes a framework integrating three cultural dimensions: community structures, environmental adaptation, and vernacular building traditions. A Situational Research Method (SRM), inspired by metamodern theory, oscillates between spatial metrics and cultural interpretation. Findings reveal distinct morphological signatures: Gharb Seheyl’s linear form supports tourism-integrated preservation; Elephantine Island’s fragmented fabric reflects millennial continuity; Heisa Island’s compact morphology embodies resilience vernacular. SRM effectively delineates socio-cultural distinctions within the urban fabric, enhancing scientific rigor in heritage research and contributing to sustainable conservation of Nubian vernacular heritage. Full article

This study examines how Saudi consumers evaluate the commodification of cultural symbols in fashion amid intensified heritage branding and symbolic market expansion. It addresses a gap in the literature on internal cultural commodification, where tensions surrounding authenticity, legitimacy, and commercialization emerge within the same cultural community rather than across clearly separate cultural groups. Drawing on a culturally grounded application of the Theory of Planned Behavior and related literature on consumer ethnocentrism and moral evaluation, the study investigates how perceived authenticity, perceived cultural appropriation, ethical sense, and consumer ethnocentrism shape attitudes toward cultural commodification and purchase intention in the Saudi fashion context. Data were collected through an Arabic-language questionnaire-based survey of Saudi consumers (N = 552) using a non-probability purposive sampling approach. The measurement model employed reflective scales adapted from prior literature and was assessed for reliability and validity. To strengthen methodological rigor, the analysis also considered common method bias diagnostics. The proposed relationships were tested using partial least squares structural equation modeling (PLS-SEM) with bootstrapping. The findings indicate that perceived authenticity is positively associated with attitudes toward cultural commodification and relates to purchase intention primarily through attitudes. Perceived cultural appropriation is negatively associated with both attitudes and purchase intention, suggesting both a direct deterrent effect and an indirect pathway via attitudes. Consumer ethnocentrism shows a negative association with purchase intention and a weaker negative association with attitudes, while its moderating role appears statistically significant but limited in magnitude. Ethical sense displays a more complex pattern, combining negative indirect effects through evaluative pathways with a positive direct association with intention, consistent with qualified rather than purely restrictive participation in symbolic consumption. The study contributes to the literature by clarifying how consumer responses to heritage-based fashion commercialization are shaped by representational, ethical, and normative evaluations in a non-Western setting. Practically, it suggests that fashion brands operating in Saudi heritage markets should manage authenticity claims, symbolic legitimacy, and appropriation risk with greater cultural and ethical sensitivity. Full article

Campylobacter spp. is one of the most common pathogens responsible for gastroenteritis in developed countries and is raising public health concerns worldwide. This work optimized a label-free electrochemical genosensor based on screen-printed gold electrodes (SPAuEs) for the rapid detection of Campylobacter jejuni, C. coli, C. lari and C. upsaliensis. SPAuEs were functionalized with a specific thiolated DNA probe and tested with a ferrocyanide solution for signal production. The optimization of the conditions was obtained using DNA extracted from pure cultures of Campylobacter spp. and negative controls such as Escherichia coli, Listeria innocua, Salmonella spp., and Helicobacter pylori. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were compared to assess sensitivity and specificity. The relative change in intensity of the ferrocyanide anodic peak (Ipa) was proportional to the value of Campylobacter spp. DNA concentrations in the range of 1 pg/µL to 10⁴ pg/µL. The limit of detection of our optimized system was 1.06 pg/μL. After optimization, the method was applied to chicken meat samples from the market. The proposed electrochemical DNA biosensor was able to detect Campylobacter jejuni, C. coli, C. lari and C. upsaliensis after selective enrichment and DNA isolation within 60 min of DNA extraction, demonstrating its usefulness for routine analyses. Full article

Insects are widely distributed across the globe and exhibit strong adaptability in diverse living environments, a capability closely linked to the diversity of their gut microbiota. The composition of insect gut bacteria varies with species, living environment, diet, and development stage. In recent years, the widespread application of culture-independent strategies based on molecular biology techniques has provided substantial information for studies on the interaction mechanisms between insects and their gut microbiota. However, culture-dependent strategies aimed at isolating pure cultures remain indispensable. Only by integrating multi-techniques such as bacterial isolation and pure culture, axenic insect technology, and molecular biology can in-depth research be conducted on key gut bacteria of insects. This review summarizes culture-dependent and -independent strategies used for the analysis of the diversity and functions of insect gut microbiota, focusing on the traditional methods and new strategies for microbial cultivation, multi-omics techniques, and axenic insect technology. Recent studies showed that the application of integrated techniques is powerful for illustrating the microbial function and evolution of gut microbiota, and the interactions between intestinal bacteria and their hosts. Studies have shown that the insect gut microbiota plays important roles in the promotion of host growth and development by regulating host metabolic pathways, contributing to host nutrition, and supporting the host in defending against pathogens or degrading toxic compounds. Future research directions and strategies are also proposed, providing insights into further exploration of the interaction mechanisms between symbiotic insect gut bacteria and their hosts, as well as future applications in various fields. Full article