PLoS ONE最新文献

The Genetically Encoded Death Indicator (GEDI) is a ratiometric, dual-fluorescence biosensor that enables real-time detection of cell death through calcium influx. Originally developed for use in neurodegeneration models, GEDI can be applied to cancer cells to quantify therapy-induced death at single-cell resolution. This protocol details how to generate GEDI-expressing cancer cell lines, empirically determine stress-induced GEDI thresholds using radiation or chemotherapeutic agents, and perform time-resolved imaging and image analysis to track cell fate. This workflow is optimized for high-throughput drug and radiation screening in heterogeneous populations and is especially useful for identifying chemo- and radio-resistant subclones. Key limitations include the need for empirical GEDI threshold calibration for each treatment condition and careful standardization of imaging parameters. The protocol outputs include GEDI ratio values, single-cell time-of-death annotations, and whole-cell morphological data in parallel, which can be linked to downstream applications such as FACS-based isolation of live or dying subpopulations, transcriptomic profiling of resistant clones, or in vivo validation using xenografts or organotypic slice culture.

Head and neck squamous cell carcinoma (HNSCC) is a significant cause of morbidity and mortality worldwide, with limited treatment options for patients with locally advanced disease. CD47 immune checkpoint inhibitors have been used to block the CD47/SIRPa interaction that inhibits antigen-presenting cell phagocytosis, thereby enhancing antigen presentation to cytotoxic T-cells, and have shown promise in combination with anti-PD1 immunotherapy in tumors, including recurrent/metastatic HNSCC. We found that CD47 expression is associated with poor prognosis in HNSCC and explored the anti-tumor activity of an anti-CD47 fusion protein in combination with anti-PD1 and lymphatic-sparing radiotherapy in a locally advanced HNSCC model. In the 4MOSC1 syngeneic HPV-negative HNSCC mouse model, ALX301 (an engineered CD47-blocking SIRPα fusion for murine models) induced complete tumor regression when combined with anti-PD-1, and produced a partial tumor response as a monotherapy. An anti-PD1 immune checkpoint inhibitor in a CD47-null tumor background led to complete tumor regression confirming a key role for CD47 in tumor immunity. ALX301 treated mice demonstrated increased MHC-II expression on dendritic cells within the tumor and upregulation of CD86 co-stimulatory molecule on dendritic cells within the tumor, sentinel lymph nodes, and contralateral lymph nodes. Combination ALX301 and anti-PD1 treatment in an anti-PD1 resistant 4MOSC2 model demonstrated significant tumor regression, enhanced survivability, improved response with neoadjuvant radiotherapy, and greater retention of CD8 + T-cells within the tumor microenvironment. Notably, T-cell receptor sequencing revealed increased shared clonality between the tumor and sentinel lymph nodes of ALX301 treated mice. These data demonstrate that a combination of CD47 blockade and anti-PD1 therapy enhances tumor antigen presentation and immune cell infiltration, while further improving anti-tumor responses in combination with tumor-targeted radiotherapy. This study provides support for the rational design of combinatorial immunoradiotherapy, using anti-CD47 inhibitors and anti-PD1 therapy, in a clinical trial targeting locally advanced HPV-negative HNSCC.

Background: Today, community pharmacists' responsibilities have expanded beyond the traditional role to include the management of minor ailments. Acute uncomplicated cystitis is one of the most prevalent medical conditions seen in primary healthcare and can be managed by community pharmacists (CPs).

Objectives: To evaluate community pharmacists' history-taking practice when responding to patients with acute uncomplicated cystitis.

Methods: A cross-sectional simulated patient study was conducted from November 2022 to January 2023 in Khartoum locality targeting only pharmacists. Six trained female students played the simulation in which they pretended to have the clinical symptoms of acute uncomplicated cystitis and requested treatment for their condition. The Interactions during the simulation were documented immediately in a data collection form after each visit. Descriptive statistics were used to report the study outcomes.

Results: A total of 238 community pharmacies were visited. The majority of the pharmacists were female. The mean of the number of questions that were asked by the pharmacists was 1 (SD = 1.6) questions. Among the studied pharmacists, 45.4% asked at least one question during patient assessment. The most asked question was if the patient has a fever, representing 61 (25.6%) CPs, followed by if there is vaginal discharge, representing 38 (16%) CPs. In response to scenario 47 (19.7%) CPs decided to refer the patient to a physician, and 45 (18.9%) of the CPs advised the patient to increase water intake.

Conclusions: The study has revealed a poor history-taking practice towards acute uncomplicated cystitis during patient assessment. Further studies exploring pharmacists' involvement in patient assessment are required. Strategies to improve community pharmacists' practice, like continuing pharmacy education and providing a national guideline regarding patient assessment should be considered seriously.

Background: Diethyl phthalate (DEP), a widely used plasticizer with endocrine-disrupting properties, has raised concerns regarding its potential carcinogenic effects. However, its precise role in colorectal cancer (CRC) development remains poorly understood.

Methods: The chemical structure of DEP was obtained from the PubChem database. Potential targets of DEP were identified through ChEMBL and STITCH databases and intersected with known CRC-related genes to screen for candidate biomarkers. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to explore the biological functions and signaling pathways involved. Molecular docking was conducted to predict the binding affinities between DEP and core targets. Finally, 200-ns molecular dynamics (MD) simulations using GROMACS were employed to evaluate the binding stability and dynamic behavior of the DEP-target complexes.

Results: A total of 62 overlapping genes were identified between DEP targets and CRC-associated genes. GO and KEGG enrichment analyses indicated enrichment in epigenetic regulation, chromatin remodeling, and cancer-related signaling pathways, including Notch, TGF-β, and FoxO. Protein-protein interaction analysis identified EP300, EZH2, HDAC1, HDAC2, and KDM1A as key epigenetic regulators. Molecular docking predicted moderate binding affinities between DEP and these targets (-6.6 to -5.7 kcal·mol ⁻ ¹). Subsequent 200-ns MD simulations suggested that DEP formed stable complexes with HDAC1, KDM1A, and EZH2, moderate stability with EP300, and partial dissociation with HDAC2, consistent with hydrophobic and hydrogen-bonding interactions at the binding interfaces.

Conclusion: This study provides a theoretical framework for exploring the molecular mechanisms through which DEP may contribute to CRC development, emphasizing the value of network toxicology in cancer research. These findings may inform future investigations into the risks of DEP exposure and support public health policy and the development of targeted therapeutic strategies.

Art engagement plays a crucial role in children's flourishing, yet there is limited understanding of how individual traits and environmental factors shape children's art engagement. Drawing on Cognitive Evaluation Theory (CET), this study examines the relationship between children's drawing self-efficacy and art engagement, as well as the moderating role of perceived social support. A cross-sectional survey was conducted with 592 children aged 10-12 in Henan Province, China, using paper-based questionnaires. The results indicated that both drawing self-efficacy and perceived social support positively predicted art engagement; however, high levels of perceived social support weakened the positive relationship between drawing self-efficacy and art engagement. These findings challenge the common assumption that social support is uniformly beneficial and highlight the importance of autonomy-supportive practices. The key challenge lies in providing support that strengthens children's autonomy in drawing activities. Parents and art educators should therefore adopt autonomy-supportive approaches to help children remain actively engaged in drawing activities and achieve long-term developmental benefits.

Grapevine Trunk Diseases (GTDs) are caused by phytopathogenic fungi that compromise grapevine productivity and wine quality. Most GTDs preventive treatments are chemical-based and environmentally harmful. One goal of the European Green Deal is to develop sustainable agriculture which does not harm the environment and reduces pesticide use and an alternative to those treatments may be the use of elicitors such as oligosaccharides from fungi. Many studies confirm that oligosaccharides activate the defence response. The experiment was carried out in vineyards of Tempranillo and Airén cvs. Asymptomatic and symptomatic vines were treated with mannans. Leaves and grapes were taken and pigments and phenols content, polyphenol oxidase (PPO) and superoxide dismutase (SOD) activities and gene expression of several defence enzymes were determined. The mannan addition to symptomatic vines was more positive for the leaves than for the grapes, palliating the damage caused by the disease, especially in the cv. Tempranillo. On the one hand, in the leaves, mannans caused an increase in phenols and PPO activity and expression; on the other hand, in grapes, although phenols increased, the other parameters did not. Mannans increased the expression levels of chalcone synthase (CHS1, CHS3), phenylalanine ammonia lyase (PAL), SOD, and PPO in asymptomatic leaves of both cultivars. In symptomatic leaves, CHS3 and PAL expression decreased in both cultivars, while CHS1 and PPO increased only in Tempranillo. In grapes, the expression of the genes varied due to the development of the disease. The mannan treatment seemed to reduce the oxidative stress caused by GTDs, but, above all, mannans would act as a biostimulant activaing the defence system of asymptomatic vines that would help them respond more successfully to a possible pathogenic fungi infection, that although this response depended on the cultivar.

Accurate mortality risk assessment is critical for decision-making in life insurance, healthcare, and public policy. Regional variability in mortality, driven by diverse local factors and inconsistent data availability, presents significant modeling challenges. This study introduces a novel hierarchical mortality risk model that integrates global and local data, enhancing regional mortality estimation across diverse regions. The proposed approach employs a two-stage process: first, a global Light Gradient Boosting Machine model is trained on globally shared features; second, region-specific models are developed to incorporate local characteristics. This framework outperforms both purely local models and standard imputation techniques, particularly in data-scarce regions, by leveraging global patterns to improve generalization. The model is computationally efficient, scalable, and robust in handling missing values, making it adaptable for other domains requiring integration of multi-regional data. This method enhances predictive accuracy across various regions and provides a more reliable approach for mortality risk estimation in data-scarce environments.

Objective: Explore the causes and mechanisms of bladder cancer-induced Cardiovascular diseases (CVD) death.

Method: We acquired bladder cancer patient data from the SEER database to evaluate CVD death risk. Cross-WGCNA was employed to identify comorbidity genes linking bladder cancer and heart failure. Functional phenotypes of bladder cancer cell lines were analyzed using cell culture, transaction, CCK-8, and Transwell assays, while ELISA determined extracellular target protein concentrations. Myocardial cell function was assessed by examining cell proliferation, collagen I levels, and mitochondrial membrane potential.

Result: Our study, analyzing 140,760 bladder cancer patients from the SEER database, revealed that CVD is a major cause of death, increasing risk by 18%. Cross-WGCNA and Lasso regression identified SFRP1 and CAPG as key serum proteins linked to bladder cancer and heart failure. Regulating these proteins' mRNA levels significantly impacts cancer proliferation, migration, and invasion. CAPG, in particular, suppresses myocardial cell function. We discovered SB525334 as a strong CAPG inhibitor in bladder cancer cells, potentially enhancing cisplatin's effectiveness by targeting CAPG.

Conclusion: Bladder cancer patients face an elevated CVD death risk due to high CAPG protein expression, which can raise serum CAPG levels and harm cardiomyocytes.

Objectives: Research on overindebtedness in general and on the relationship between overindebtedness and being overweight or obese in particular is extremely rare or practically nonexistent although both phenomena have shown an increasing trend in recent years and are expected and found to be more prevalent among lower social classes and educational levels. However, no such study for Switzerland has ever been conducted until now.

Methods: Survey data collected in 2019 from 219 overindebted adult clients of four official debt advisory centers in the Canton of Zurich were used and linked with a sample of 1,997 respondents of the Swiss Health Survey 2017 of the same age and canton of residence. The entire study population included a total of 2,216 adult individuals. Contingency tables with relative frequencies were calculated to study differences between the two subsamples. Furthermore, logistic and Poisson regression analyses were performed to calculate unadjusted and multiple-adjusted odds ratios and risk ratios as proxies and measures for the relative risk of being overweight or obese among overindebted people.

Results: The prevalence rates of being overweight and having a body mass index (BMI) of 25+ and particularly of being obese (BMI of 30+) were significantly higher among overindebted individuals (BMI ≥ 25: 46%, BMI ≥ 30: 15%) than predominantly non-overindebted people (BMI ≥ 25: 38%, BMI ≥ 30: 9%). Overindebtedness increased the odds or the relative risk for such unfavourable body weights by 20% to 36% (overweight) and by 59% to 70% (obesity) depending on the effect measure considered. This was found regardless of overindebted individuals' sex, age and educational level and independent of the fact that they have a comparably very low sense of control, feel lonely much more often and show much more often moderate to severe depressive symptoms.

Conclusions: Measures of effect or association found were statistically significant at least for obesity, but smaller than expected and somewhat under- and simultaneously overestimated in view of the younger average age and the lower educational level of the overindebted individuals.