Search Results (51)

Herein, we report a simple procedure regarding the photodynamic therapy (PDT) treatment as a minimally invasive modality for treating superficial bladder cancer that utilizes a photosensitizer, light, and oxygen to generate cytotoxic reactive oxygen species (ROS). This study evaluates the histopathological and morphological changes induced by PDT in an ex vivo model of low-grade (LG) pTa non-muscle-invasive bladder cancer (NMIBC). We investigated the efficacy of exogenous protoporphyrin IX (PpIX) and Rose Bengal (RB) by incubating tissue samples (n = 30) with an oxygen-saturated solution of PpIX (1–3 mM) or RB (0.3–0.5 mM) for one hour. Since the criticism of using frozen tissue in research already exists, this framing explains how to mitigate those limitations. Thus, we use oxygen-saturated solutions PpIX and oxygen-saturated solutions of RB. We discussed a few aspects related to the use of frozen tissue in PDT. Frozen tissue preserves lipids critical for assessing membrane damage and maintains higher levels of metabolic markers like antioxidant molecules like glutathione and more likely lack factors such as metabolic activity, intact cell membranes, and oxygenation. It is critical to differentiate between “artifactual” changes and the “pathological” death of cells. Thus, we used histopathological microscopy observation typically used in daily clinical investigations to characterize cells before and after PDT. Following irradiation with the light dose of 72 J/cm² (410 nm or 532 nm at 300 mW for 15 min), hematoxylin–eosin staining revealed concentration-dependent apoptotic changes, including chromatin condensation, pyknosis, and nuclear fragmentation. While both agents induced cell death, RB demonstrated faster and more intense cytotoxicity than PpIX. These findings provide microscopic evidence of PDT-induced tumor destruction and suggest that RB is a potent candidate for further preclinical evaluation. At 410 nm (deep blue/violet), light penetration in biological tissue is very shallow, typically only around 0.3 to 1 mm; therefore, in a 2 mm thick tissue sample, most of the light would be absorbed within the first millimeter, with minimal light reaching the full depth of tissues. In this protocol, the generated ROS is used to destroy tumor tissue by attacking the cellular microenvironment directly. This led to immediate membrane disruption and lipid peroxidation. The proof-of-concept is an early-stage study designed to verify that a PDT treatment is feasible, safe, and biologically active in an ex vivo model of LG pTa NMIBC. Full article

Background: Untargeted metabolomics enables comprehensive profiling of biological systems, but accurate metabolite annotation remains a critical bottleneck due to incomplete spectral libraries and structural isomerism. The use of in silico annotation tools can increase the coverage of annotated compounds, but it remains unclear whether these tools, in the absence of reference standards, can reliably annotate real-world experimental LC-HRMS data and whether they are sufficient for this task. Methods: This study assesses the performance and limitations of four widely used in silico structure prediction tools (MassFrontier, MetFrag, MS-FINDER, and SIRIUS/CSI:FingerID) when applied to an experimentally acquired feature set previously used to differentiate patients with depressive disorders from healthy controls. To ensure uniform evaluation across tools under realistic but optimized conditions, the quality of MS/MS data was improved using a parallel reaction monitoring method, allowing acquisition of interpretable fragmentation spectra for 26 of the 28 detected features. Results: For most features, all tools were able to suggest structure candidates. However, none of the tools proved sufficient as a standalone solution for reliable metabolite annotation. Due to their different algorithms, each tool had strengths and weaknesses in fragmentation interpretation, candidate generation, and ranking, resulting in incomplete or inconsistent annotations. While the combined application of all four tools provided a substantial improvement in putative annotation over conventional spectral library matching, the in silico structure prediction tools often prioritized chemically implausible, biologically irrelevant, or artifactual candidates. Consequently, manual expert evaluation was required to assess the chemical plausibility and biological relevance of the proposed structures. This ultimately reduced the number of biologically plausible metabolites putatively associated with disease to ten. Conclusions: Overall, these results demonstrate that existing in silico annotation tools can substantially support the annotation of experimental metabolomics data, but are insufficient on their own. Reliable identification of metabolites in complex biological matrices still depends on high-quality MS/MS data acquisition, the combined use of complementary tools, and mandatory post-annotation expert curation. Full article

This study introduces an innovative hybrid framework for the sustainable design of meditation centers by integrating entropy-weighted fuzzy comprehensive evaluation (FCE) with principles of cultural sustainability. While conventional sustainable design assessment methods predominantly emphasize technical environmental performance, they remain insufficient for meditation center design, where contemplative experience, cultural continuity, and spatial meaning are equally essential. In response to this gap, this research reinterprets the Mogao Caves as an exemplar of “deep sustainability,” where environmental, social, and cultural dimensions are integrated in a mutually reinforcing manner. Through a systematic analysis of the spatial and artifactual heritage of the Mogao Caves, a robust and quantifiable evaluation system consisting of 27 indicators was developed, spanning architectural design, spatial organization, seating iconography depicted in the murals, and decorative elements. The novelty of this study lies in establishing a heritage-informed and data-driven framework that translates historical spatial wisdom into a contemporary sustainable design assessment model. By applying the entropy-weighting method, the study identifies Functional Diversity (0.087) and Symbolic Representation (0.071) as indicators with comparatively greater discriminative contribution within the present sample, highlighting the importance of programmatic adaptability and cultural expression in meditation center design. The FCE model was applied to 156 valid questionnaire responses, enabling a multi-criteria evaluation of 11 meditation centers worldwide, among which the Fujian Longyan Dahe Meditation Center achieved the highest score (73.032). The findings indicate that the proposed framework offers a more balanced basis for evaluating meditation center design by integrating functional performance with cultural continuity and spatial meaning. Full article

When measurement invariance (MI) is violated in multi-group structural equation models, group-specific measurement artifacts inflate the between-group variance of structural parameters beyond their true values. Existing remedies—partial invariance, group-specific estimation, or moderation analysis—address the consequences of inflation but not its mechanism. This article introduces the stabilizer variable, a covariate that absorbs measurement-induced parameter heterogeneity while maintaining structural independence from the focal relationship. Two theoretical results are established: a variance decomposition theorem showing that MI violations inflate dispersion through an identifiable artifactual component, and a purification theorem proving that a stabilizer reduces this dispersion via Frisch–Waugh–Lovell projection. Two stabilization mechanisms are identified: variance purification (Type A) and directional alignment (Type B). We then develop the stabilizer variable test, a dual-criterion procedure combining nonparametric bootstrap testing for stabilization magnitude with binomial testing for directional consistency, incorporating adaptive MI severity scoring with calibrated fit-index weights. Simulations comprising 949,100 replications across varying group counts, sample sizes, and MI severity levels demonstrate 80–99% power with false-positive rates below 2%. Practical guidelines recommend $K\ge 10$ groups and $n\ge 100$ per group for conservative applications. The framework generalizes to any multi-group regression context where systematic measurement error induces spurious parameter heterogeneity. Full article

Over the past two decades, numerous studies have examined the etiological significance of DNA fragmentation in human sperm using methods such as the comet assay (CA), the sperm chromatin structure assay, the sperm chromatin dispersion assay, and the TUNEL assay. We developed single-cell pulsed-field gel electrophoresis techniques, including one-dimensional (1D-SCPFGE) and angle-modulated two-dimensional (2D-SCPFGE), to detect early signs of naturally occurring DNA fragmentation. Comparative studies using purified human sperm with and without DNA fragmentation revealed some technical limitations in the conventional methods. This technical review outlines the procedures to ensure the quantitative performance of SCPFGE: (1) The mass of naked DNA was prepared through simultaneous in-gel swelling and proteolysis, which are highly sensitive to chemical and physical factors. Notably, these processes are vulnerable to reactive oxygen species (ROS). We developed the anti-ROS SCPFGE system to prevent artifactual cleavages. (2) 1D-SCPFGE discharges long-chain fibers from the origin, separating fibrous and granular segments beyond the tips of the fibers. (3) During continuous electrophoresis after 150° rotation (2D-SCPFGE-0-150), long-chain fibers unexpectedly extended diagonally backward from the origin, with long fibrous segments pulled out from a bundle that extended during the first electrophoresis, indicating some fibrous segments were embedded within the long-chain fibers. Even when SCPFGE was employed, one-directional current led to false negatives. (4) 2D-SCPFGE with angle rotation is currently the most sensitive imaging method for single-nuclear DNA fibers. However, without knowing the size of DNA fragments, it remains a semi-quantitative analysis. (5) To prevent artifactual DNA cleavage caused by ice crystals, low-temperature liquid storage is recommended. (6) The in-gel proteolyzed naked DNA is suitable as a substrate for chemical and enzymatic DNA cleavage analyses. Full article

Background/Objectives: Drug-induced sleep endoscopy (DISE) is used in obstructive sleep apnea (OSA) to visualize dynamic upper airway collapse, but sedation protocols vary widely with no consensus on the optimal agent or technique. This narrative review aims to clarify current sedation strategies for DISE in OSA and their clinical implications. Methods: We systematically searched PubMed, Scopus, Web of Science, and Cochrane Library for English-language publications on DISE sedation (2000–2025). Relevant clinical studies, guidelines, and reviews were included. Data were qualitatively synthesized due to heterogeneity among studies. Results: Sedation approaches in DISE varied considerably. Propofol, dexmedetomidine, and midazolam were the primary agents identified. Propofol provided rapid, titratable sedation but increased airway collapsibility at higher doses; dexmedetomidine produced a more natural sleep-like state with minimal respiratory depression; midazolam was less favored due to prolonged effects. Use of target-controlled infusion (TCI) and pharmacokinetic–pharmacodynamic (PK–PD) models improved control of propofol sedation. Co-sedative adjuncts (e.g., opioids) reduced the required sedative dose but added risk of respiratory depression. Careful titration to the lowest effective dose-often guided by bispectral index (BIS) monitoring—was emphasized to achieve adequate sedation without artifactual airway collapse. No universal DISE sedation protocol was identified. Conclusions: Optimal DISE sedation balances adequate depth with patient safety to ensure reliable findings. Using the minimum effective dose, guided by objective monitoring (e.g., BIS), is recommended. There is a need for standardized sedation protocols and further research (e.g., in obese patients) to resolve current controversies and improve DISE’s utility in OSA management. Full article

In this paper, we consider the inevitable large fluctuations of pressure in typical molecular dynamics (MD) simulations of ligand–protein binding problems. In simulations under the constant pressure of one bar, the pressure artifactually fluctuates over the range of $\pm 100$ bars or more. This artifact can cause gross inaccuracy in the apparent binding affinity computed as the ratio of the probability for the ligand to be bound inside the protein and the probability for the ligand to be outside the protein. Based on statistical thermodynamics, we derive a correction factor for the ligand–protein binding affinity to compensate for the artifactual pressure fluctuations. The correction factor depends on the change in the system volume between the bound and the unbound states of the ligand. We conducted four sets of MD simulations for glycerol affinities with four aquaglyceroporins: AQP10, AQP3, AQP7, and GlpF. Without the correction factor, the apparent affinity of glycerol with each of these four aquaglyceroporins is computed directly from the simulations to be very low (~1/M). With the correction factor applied, glycerol’s affinity is computed to be 1/mM to 1/µM. In conclusion, glycerol has high affinity for its native facilitator aquaglyceroporins, which is in contrast to the current literature not correcting the artifactual consequences of the large pressure fluctuations in typical in silico experiments. Full article

This article proposes the term ‘generative artifact’ to define a new method of imagining the future, one derived from artistic and architectural interpretations of non-linear time, material exploration, and relationship building. This contrasts the imagining that happened in the past by European and North American dominant culture, born out of fears of a declining Western hegemony and resulting in socially constructed hierarchies based on race. To investigate this historic and outdated imagining of culture, we trace the history of Chinatown and the ornamented feminine body as a physical example of hypervisibility in the North American city. First, we examine the current discourse on Chinatowns’ Orientalist aesthetics, legitimacy through institutionalized nonspecificity, and architectural/artifactual heritage, which serve as a mirror and moor for the Chinese diaspora today. Here, we find clues on how to navigate and leverage the spectacle of the racial image, the continuous merging of person and thing, and the tropes that the racialized body might find itself answering for. To illustrate the potential of the generative process and through the lenses of Anne Anlin Cheng’s theory of ornamentalism and Legacy Russell’s glitch feminism, this article places Chinatown adjacent to the worldbuilding and artistic practices of seven contemporary artists and architects. This includes Astria Suparak (performance critique), Curry J. Hackett (AI, installation), Shellie Zhang (sculpture), Lan “Florence” Yee (textile), Debra Sparrow (weaving, murals), Thomas Cannell (sculpture), and the author (performance). All are from varied cultural backgrounds who create ‘generative artifacts’ in their creative practices—works that playfully slip between sign/icon, high/low tech, and authentic/invented culture to point towards a path to imagining more expansive futures. Full article

Background: Hypercalcemia is a common and serious complication of malignancy, often contributing to morbidity and mortality. In patients with paraproteinemia, elevated total calcium with normal ionized calcium, termed pseudohypercalcemia, can complicate diagnosis and lead to inappropriate treatment. While this phenomenon has been described in case reports, its prevalence and clinical impact in routine practice remain poorly defined. Methods: We report a case of pseudohypercalcemia in a patient with IgG κ multiple myeloma and conducted a retrospective review of de-identified data to assess the prevalence and biochemical associations of pseudohypercalcemia in paraproteinemia. Available data included serum protein electrophoresis (SPEP), total calcium, albumin, total protein, creatinine, and parathyroid hormone (PTH). Associations between calcium status, paraprotein levels, and the gamma globulin gap were examined. Results: The index case demonstrated pseudohypercalcemia, with elevated total calcium (13.5 mg/dL) but normal ionized calcium (1.22 mmol/L), in the setting of IgG κ paraproteinemia (4.4 g/dL). In the retrospective cohort of 2537 samples, 986 (39%) had a single monoclonal paraprotein. Gamma globulin gap showed a moderate correlation with paraprotein concentration for IgG (r = 0.56, p < 0.0001) and IgA (r = 0.44, p < 0.0001), but a weaker relationship for IgM (r = 0.49, p < 0.0001). In contrast, total calcium showed no significant correlation with paraprotein concentration in the overall cohort. Among samples with elevated calcium (>10.5 mg/dL), the association between calcium and IgG paraprotein levels remained weak (r = 0.34, p = 0.23), and was similar for IgG κ (r = 0.61, p = 0.12) and IgG λ (r = 0.09, p = 0.87). Hypercalcemia was uncommon, occurring in only ~2% of IgG-positive samples, and rarely at paraprotein levels ≥ 1.5 g/dL. Conclusions: Pseudohypercalcemia in paraproteinemia is uncommon but clinically important, as total calcium may be artifactually elevated due to paraprotein-related assay interference, either from assay precipitation effects or calcium binding by paraproteins. Paraprotein burden correlates with gamma globulin gap but not with true calcium status. Reliance on total calcium alone may lead to diagnostic misclassification; ionized calcium should be measured in patients with monoclonal gammopathies to distinguish true hypercalcemia from analytical interference and avoid unnecessary treatment. Full article

Studies of ancient Israelite religion have long assumed that texts played some role in its public expression. This role is often reconstructed using depictions in the Hebrew Bible and ritual texts from neighboring regions or the Bronze Age Levant. However, no such ritual texts have been uncovered in the Iron Age Levant. Nevertheless, an analysis of architecturally embedded texts alongside their associated assemblages makes it possible to reconstruct ancient Levantine ritual practices and the roles of texts within them. As components of built environments, texts drew attention to particular areas, directing traffic along particular routes and halting it at waypoints. Texts of various genres occasionally prescribe specific ritual actions to carry out at these waypoints. Even texts lacking prescriptions were often accompanied by iconography depicting ritual practices or functional artifacts implying them. Analyzing architectural, textual, iconographic, and artifactual evidence together allows us to reconstruct ritual sequences performed in ancient built environments. This article demonstrates this method using case studies derived from four Iron Age Levantine sites: Karatepe, Karkemish, Kuntillet ʿAjrud, and Deir ʿAlla. Full article

Bilirubin is a key component in the preparation of two traditional Chinese medicines: Calculus bovis sativus and Calculus bovis artifactus. Currently, industrial-scale production of bilirubin is limited to extraction from pig bile in a very low yield and its market price is very high, so it is important to develop an alternative method for producing bilirubin. This study developed a potential process for bilirubin production through biotransformation of biliverdin. The codon-optimized gene for biliverdin reductase (BVR) from Synechocystis PCC6803 was recombinantly expressed in Komagataella phaffii GS115, resulting in the genetically modified strain GS115-bvdR, which successfully expressed BVR with intracellular activity. Whole cells of GS115-bvdR were capable of transforming biliverdin to bilirubin in vitro. The overexpression conditions were optimized to enhance BVR production by GS115-bvdR, and the optimal conditions for the biotransformation of biliverdin into bilirubin using resting GS115-bvdR cells were established (pH 5.0 buffer, at 30 °C for 24 h, with 200 mg/L biliverdin). Under these conditions, a bilirubin concentration of 153 mg/L was achieved, with a conversion of 76.2% from biliverdin. These findings provide valuable insights for future studies on the biosynthesis of bilirubin through metabolic engineering. Full article

Recent work in corpus linguistics has observed that informativity predicts articulatory reduction of a linguistic unit above and beyond the unit’s predictability in the local context, i.e., the unit’s probability given the current context. Informativity of a unit is the inverse of average (log-scaled) predictability and corresponds to its information content. Research in the field has interpreted effects of informativity as speakers being sensitive to the information content of a unit in deciding how much effort to put into pronouncing it or as accumulation of memories of pronunciation details in long-term memory representations. However, average predictability can improve the estimate of local predictability of a unit above and beyond the observed predictability in that context, especially when that context is rare. Therefore, informativity can contribute to explaining variance in a dependent variable like reduction above and beyond local predictability simply because informativity improves the (inherently noisy) estimate of local predictability. This paper shows how to estimate the proportion of an observed informativity effect that is likely to be artifactual, due entirely to informativity improving the estimates of predictability, via simulation. The proposed simulation approach can be used to investigate whether an effect of informativity is likely to be real, under the assumption that corpus probabilities are an unbiased estimate of probabilities driving reduction behavior, and how much of it is likely to be due to noise in predictability estimates, in any real dataset. Full article

Background: Genetic variation provides a foundation for understanding evolution. With the rise of artificial intelligence, machine learning has emerged as a powerful tool for identifying genomic footprints of evolutionary processes through simulation-based predictive modeling. However, existing approaches require prior knowledge of the factors shaping genetic variation, whereas uncovering anomalous genomic regions regardless of their causes remains an equally important and complementary endeavor. Methods: To address this problem, we introduce ANDES (ANomaly DEtection using Summary statistics), a suite of algorithms that apply statistical techniques to extract features for unsupervised anomaly detection. A key innovation of ANDES is its ability to account for autocovariation due to linkage disequilibrium by fitting curves to contiguous windows and computing their first and second derivatives, thereby capturing the “velocity” and “acceleration” of genetic variation. These features are then used to train models that flag biologically significant or artifactual regions. Results: Application to human genomic data demonstrates that ANDES successfully detects anomalous regions that colocalize with genes under positive or balancing selection. Moreover, these analyses reveal a non-uniform distribution of anomalies, which are enriched in specific autosomes, intergenic regions, introns, and regions with low GC content, repetitive sequences, and poor mappability. Conclusions: ANDES thus offers a novel, model-agnostic framework for uncovering anomalous genomic regions in both model and non-model organisms. Full article

The assessment of ocular surface temperature (OST) plays a pivotal role in the diagnosis and management of various ocular diseases. This paper introduces significant enhancements to the ThermOcular system, initially developed for precise OST measurement using infrared (IR) thermography. These advancements focus on accuracy improvements that reduce user dependency and increase the system’s diagnostic capabilities. A novel addition to the system includes the use of EyeTags, which assist clinicians in selecting control points more easily, thus reducing errors associated with manual selection. Furthermore, the integration of state-of-the-art semantic segmentation models trained on the newest dataset is explored. Among these, the OCRNet-HRNet-w18 model achieved a segmentation accuracy of 96.21% MIOU, highlighting the effectiveness of the improved pipeline. Additionally, the challenge of eliminating eyelashes in IR frames, which cause artifactual measurement errors in OST assessments, is addressed. Through a newly developed method, the influence of eyelashes is eliminated, thereby enhancing the precision of temperature readings. Moreover, an algorithm for blink detection and elimination is implemented, significantly improving upon the basic methods previously utilized. These innovations not only enhance the reliability of OST measurements, but also contribute to the system’s efficiency and diagnostic accuracy, marking a significant step forward in ocular health monitoring and diagnostics. Full article

Radars are promising tools for contactless vital sign monitoring. As a screening device, radars could supplement polysomnography, the gold standard in sleep medicine. When the radar is placed lateral to the person, vital signs can be extracted simultaneously from multiple body parts. Here, we present a method to select every available breathing and heartbeat signal, instead of selecting only one optimal signal. Using multiple concurrent signals can enhance vital rate robustness and accuracy. We built an algorithm based on persistence diagrams, a modern tool for time series analysis from the field of topological data analysis. Multiple criteria were evaluated on the persistence diagrams to detect breathing and heartbeat signals. We tested the feasibility of the method on simultaneous overnight radar and polysomnography recordings from six healthy participants. Compared against single bin selection, multiple selection lead to improved accuracy for both breathing (mean absolute error: 0.29 vs. 0.20 breaths per minute) and heart rate (mean absolute error: 1.97 vs. 0.66 beats per minute). Additionally, fewer artifactual segments were selected. Furthermore, the distribution of chosen vital signs along the body aligned with basic physiological assumptions. In conclusion, contactless vital sign monitoring could benefit from the improved accuracy achieved by multiple selection. The distribution of vital signs along the body could provide additional information for sleep monitoring. Full article