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14 pages, 1804 KiB  
Article
Neurotoxicity and Mechanism in Zebrafish Embryo Induced by Tetrabromobisphenol A bis (2-Hydroxyethyl) Ether (TBBPA-DHEE) Exposure
by Xinyu Zhang, Liguo Guo, Yiwen Luo, Xia Xu, Ying Han, Hui Chen, Haohao Sun, Yingang Xue and Guixiang Ji
Toxics 2025, 13(2), 76; https://doi.org/10.3390/toxics13020076 - 22 Jan 2025
Abstract
Tetrabromobisphenol A bis (2-hydroxyethyl) ether (TBBPA-DHEE), a derivative of TBBPA, has been frequently detected in the environment. In this study, the median lethal concentration (LC50) of TBBPA-DHEE at 96 h post-fertilization (hpf) was 1.573 mg/L. Based on the reported environmental concentrations, [...] Read more.
Tetrabromobisphenol A bis (2-hydroxyethyl) ether (TBBPA-DHEE), a derivative of TBBPA, has been frequently detected in the environment. In this study, the median lethal concentration (LC50) of TBBPA-DHEE at 96 h post-fertilization (hpf) was 1.573 mg/L. Based on the reported environmental concentrations, we investigated the effects of TBBPA-DHEE on the nervous system of zebrafish embryos following exposure to varying concentrations (0, 20, 100, and 500 μg/L) for 4 to 144 hpf. Our results indicated that exposure to 100 μg/L at 144 hpf led to behavioral abnormalities in zebrafish. Furthermore, exposure to TBBPA-DHEE inhibited the development of the central nervous system and motor neurons in zebrafish. Real-time polymerase chain reaction (PCR) analysis revealed that exposure to TBBPA-DHEE significantly downregulated the expression levels of neurodevelopmental genes (shha, syn2a, elavl3, gfap, and gap43). Additionally, TBBPA-DHEE increased oxidative stress in zebrafish. Transcriptomic analysis demonstrated that exposure to TBBPA-DHEE affected the signaling pathways involved in neurodevelopment. Overall, this study demonstrated that TBBPA-DHEE may disrupt the early development of the nervous system, leading to abnormal motor behavior in zebrafish larvae, and provided novel insights into the potential mechanisms of TBBPA-DHEE neurotoxicity. Full article
(This article belongs to the Special Issue Neuronal Injury and Disease Induced by Environmental Toxicants)
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Graphical abstract

Graphical abstract
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<p>Effects of TBBPA-DHEE on the zebrafish embryos and larvae’s development index. (<b>A</b>) Fitted curves of survival of zebrafish larvae exposure to TBBPA-DHEE at 96 hpf at concentration gradients (0.125 mg/L, 0.25 mg/L, 0.5 mg/L, 1 mg/L, 2 mg/L, 4 mg/L); (<b>B</b>) The zebrafish larvae hatching rate following TBBPA-DHEE exposure at 48 and 72 hpf; (<b>C</b>) The typical figure of zebrafish larvae at 72 and 144 hpf; (<b>D</b>) The yolk sac area statistical chart of zebrafish embryo at 24 hpf; (<b>E</b>) The heart rate statistical chart of zebrafish larvae at 72 hpf; (<b>F</b>,<b>G</b>) The statistical chart of bladder size and body length at 144hpf; (<b>H</b>–<b>L</b>) The exemplary locomotion tracks, distance traveled per minute, total distance, average speed and dull time after TBBPA-DHEE exposure in 144 hpf larva zebrafish. (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 and *** <span class="html-italic">p</span> &lt; 0.001).</p>
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<p>TBBPA-DHEE exposure hampered the development of neurons in zebrafish larvae. (<b>A</b>) The typical imagine of <span class="html-italic">Tg (Hb9: eGFP)</span> transgenic zebrafish following TBBPA-DHEE exposure; (<b>B</b>,<b>C</b>) The statistical graph of motor neuron length in zebrafish larvae at 72 and 144 hpf; (<b>D</b>) The typical imagine of <span class="html-italic">Tg (Gad1b: mCherry)</span> transgenic zebrafish following TBBPA-DHEE exposure; (<b>E</b>) The statistical chart of the GAD67-positive GABAergic neurons’ fluorescence area; (<b>F</b>) The impact of TBBPA-DHEE on the transcription of genes associated to neurodevelopment, including <span class="html-italic">shha</span>, <span class="html-italic">syn2a</span>, <span class="html-italic">elval3</span>, <span class="html-italic">gfap</span> and <span class="html-italic">gap43</span>. (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 and *** <span class="html-italic">p</span> &lt; 0.001).</p>
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<p>TBBPA-DHEE induces oxidative stress. (<b>A</b>) The typical fluorescence imagines of ROS in zebrafish larvae using the DCFH-DA fluorescence probe in difference treatment groups; (<b>B</b>) The statistical graph of ROS fluorescence; (<b>C</b>–<b>E</b>) The activity of CAT, SOD and the MDA content in 144 hpf zebrafish larvae (<span class="html-italic">n</span> = 50 per group, * <span class="html-italic">p</span> &lt; 0.05 and *** <span class="html-italic">p</span> &lt; 0.001).</p>
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<p>Transcriptome analysis to elucidate the toxicity of neuron development after TBBPA-DHEE exposure. (<b>A</b>) The number of DEGs up- and down-regulated between TBBPA-DHEE exposure treatments; (<b>B</b>) The top 30 GO terms. (<b>C</b>) The KEGG enrichment pathways, Spliceosome, Steroid hormone biosynthesis and ABC transporters were the most significantly enriched pathways.</p>
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32 pages, 2809 KiB  
Review
Review of Emerging and Nonconventional Analytical Techniques for Per- and Polyfluoroalkyl Substances (PFAS): Application for Risk Assessment
by Andrew McQueen, Ashley Kimble, Paige Krupa, Anna Longwell, Alyssa Calomeni-Eck and David Moore
Water 2025, 17(3), 303; https://doi.org/10.3390/w17030303 - 22 Jan 2025
Abstract
Per- and polyfluoroalkyl substances (PFAS) are persistent environmental contaminants that pose significant risks to ecosystems and human health. Increasing regulatory demands for PFAS management have increased the need for rapid and deployable analytical technologies for both abiotic and biotic matrices. Traditional detection methods, [...] Read more.
Per- and polyfluoroalkyl substances (PFAS) are persistent environmental contaminants that pose significant risks to ecosystems and human health. Increasing regulatory demands for PFAS management have increased the need for rapid and deployable analytical technologies for both abiotic and biotic matrices. Traditional detection methods, such as standardized chromatography, often require weeks to months for analysis due to a limited number of appropriately accredited laboratories, delaying critical decision-making. This literature review is intended to identify promising emerging PFAS analytical techniques or technologies to facilitate more rapid (near real-time) analysis and explore their relevancy in supporting human and ecological risk assessments. Recently developed optical and electrochemical sensing approaches are enabling the detection of PFASs within minutes to hours, with detection limits typically aligning within reported ambient concentrations in water, soil, and sediment. These emerging technologies could (1) support planning and prioritization of sampling efforts during the problem formulation phase of risk assessment, (2) complement traditional chromatography methods to lower time and resource demands to improve sampling frequency over space and time, and (3) aid in risk-informed characterization of PFAS exposures based on identified chemical classes or groups. This review highlights those approaches and technologies that could potentially enhance the comprehensiveness and efficiency of PFAS risk assessment across diverse environmental settings in the future. Full article
(This article belongs to the Section Water Quality and Contamination)
15 pages, 2394 KiB  
Review
Resilience to Global Health Challenges Through Nutritional Gut Microbiome Modulation
by Erika Isolauri and Kirsi Laitinen
Nutrients 2025, 17(3), 396; https://doi.org/10.3390/nu17030396 - 22 Jan 2025
Abstract
As the world faces an escalating challenge of non-communicable diseases (NCDs), with phenotypes ranging from allergic chronic immuno-inflammatory diseases to neuropsychiatric disorders, it becomes evident that their seeds are sown during the early stages of life. Furthermore, within only a few decades, human [...] Read more.
As the world faces an escalating challenge of non-communicable diseases (NCDs), with phenotypes ranging from allergic chronic immuno-inflammatory diseases to neuropsychiatric disorders, it becomes evident that their seeds are sown during the early stages of life. Furthermore, within only a few decades, human obesity has reached epidemic proportions and now represents the most serious public health challenge of our time. Recent demonstrations that a growing number of these conditions are linked to aberrant gut microbiota composition and function have evoked active scientific interest in host-microbe crosstalk, characterizing and modulating the gut microbiota in at-risk circumstances. These efforts appear particularly justified during the most critical period of developmental plasticity when the child’s immune, metabolic, and microbiological constitutions lend themselves to long-term adjustment. Pregnancy and early infancy epitomize an ideal developmental juncture for preventive measures aiming to reduce the risk of NCDs; by promoting the health of pregnant and lactating women today, the health of the next generation(s) may be successfully improved. The perfect tools for this initiative derive from the earliest and most massive source of environmental exposures, namely the microbiome and nutrition, due to their fundamental interactions in the function of the host immune and metabolic maturation. Full article
(This article belongs to the Special Issue Maternal Diet, Body Composition and Offspring Health)
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Figure 1
<p>The period of the first 1000 days, from pregnancy throughout infancy, offers a promising time window for interventions aiming to improve the long-term health of the child. The current evidence from clinical trials demonstrates a wide range of potential targets for microbiota modulation during this continuum (green color), while that of exposures and nutritional interventions beyond this period has been limited (red color). The blue arrow points to the intergenerational aspect of microbiome development and the risk of non-communicable diseases.</p>
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<p>Mechanistic notions on diet-gut microbiota interactions (Modified from [<a href="#B40-nutrients-17-00396" class="html-bibr">40</a>]). The current evidence from clinical trials demonstrates the action of probiotics in the gut lumen, gut barrier function, and the mucosal immune system (green color), while the evidence of the impact beyond the gut is limited (red color).</p>
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10 pages, 811 KiB  
Article
Association of Cardiovascular Disease Mortality and Ambient Temperature Variation in Shanghai, China: Beyond Air Quality Index PM2.5
by Qi Li, Shizhen Li, Ting Zhai, Shan Jin, Chunfang Wang, Bo Fang and Tian Xia
Atmosphere 2025, 16(2), 119; https://doi.org/10.3390/atmos16020119 - 22 Jan 2025
Abstract
Evidence from megacity registry data regarding the independent association between ambient temperature and cardiovascular disease (CVD) mortality, after accounting for Particulate Matter 2.5 (PM2.5), remains scarce. In this study, we collected 308,116 CVD mortality cases in Shanghai from 2015 to 2020. [...] Read more.
Evidence from megacity registry data regarding the independent association between ambient temperature and cardiovascular disease (CVD) mortality, after accounting for Particulate Matter 2.5 (PM2.5), remains scarce. In this study, we collected 308,116 CVD mortality cases in Shanghai from 2015 to 2020. The distributed lag non-linear model (DLNM) was utilized. The daily PM2.5 concentration was transformed using a natural spline (ns) function and integrated into the model for adjustment. The DLNM analysis revealed that the exposure–response curve between daily temperature and CVD mortality approximated an inverted “J” shape, consistent for both women and men. The minimum mortality temperature (MMT) for total CVD mortality was 25 °C, with an MMT of 26 °C for females and 24 °C for males. The highest relative risk (RR) of CVD mortality was 2.424 [95% confidence interval (95% CI): 2.035, 2.887] at the lowest temperature of −6.1 °C, with 2.244 (95% CI: 1.787, 2.818) for female and 2.642 (95% CI: 2.100, 3.326) for male. High temperatures exert acute and short-term effects, with the peak risk occurring on the day of exposure. In contrast, the risk from low temperature peaks on day 3 of the lag time and subsequently declines until days 16–21. This study offers evidence-based support for the prevention of temperature-induced CVD mortality. Full article
(This article belongs to the Special Issue Urban Air Pollution Exposure and Health Vulnerability)
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Figure 1

Figure 1
<p>Time series plot of daily CVD mortality and changes in temperature and humidity in Shanghai from 2015 to 2020. (<b>A</b>) Daily CVD deaths in Shanghai, 2015–2020; (<b>B</b>) Daily average temperature in Shanghai, 2015–2020; (<b>C</b>) Daily average PM<sub>2.5</sub> concentrations in Shanghai, 2015–2020.</p>
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<p>Association curve between daily average temperature and CVD deaths in Shanghai, 2015–2020. (<b>A</b>) Total CVD deaths; (<b>B</b>) CVD deaths in females; (<b>C</b>) CVD deaths in males. The grey shaded area represents the 95% confidence interval (95% CI) of the relative risk (RR) values.</p>
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16 pages, 15770 KiB  
Article
Enhancing Mixed Traffic Flow with Platoon Control and Lane Management for Connected and Autonomous Vehicles
by Yichuan Peng, Danyang Liu, Shubo Wu, Xiaoxue Yang, Yinsong Wang and Yajie Zou
Sensors 2025, 25(3), 644; https://doi.org/10.3390/s25030644 - 22 Jan 2025
Abstract
As autonomous driving technology advances, connected and autonomous vehicles (CAVs) will coexist with human-driven vehicles (HDVs) for an extended period. The deployment of CAVs will alter traffic flow characteristics, making it crucial to investigate their impacts on mixed traffic. This study develops a [...] Read more.
As autonomous driving technology advances, connected and autonomous vehicles (CAVs) will coexist with human-driven vehicles (HDVs) for an extended period. The deployment of CAVs will alter traffic flow characteristics, making it crucial to investigate their impacts on mixed traffic. This study develops a hybrid control framework that integrates a platoon control strategy based on the “catch-up” mechanism with lane management for CAVs. The impacts of the proposed hybrid control framework on mixed traffic flow are evaluated through a series of macroscopic simulations, focusing on fundamental diagrams, traffic oscillations, and safety. The results illustrate a notable increase in road capacity with the rising market penetration rate (MPR) of CAVs, with significant improvements under the hybrid control framework, particularly at high MPRs. Additionally, traffic oscillations are mitigated, reducing shockwave propagation and enhancing efficiency under the hybrid control framework. Four surrogate safety measures, namely time to collision (TTC), criticality index function (CIF), deceleration rate to avoid a crash (DRAC), and total exposure time (TET), are utilized to evaluate traffic safety. The results indicate that collision risk is significantly reduced at high MPRs. The findings of this study provide valuable insights into the deployment of CAVs, using control strategies to improve mixed traffic flow operations. Full article
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Figure 1
<p>Schematic diagram of the road segment.</p>
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<p>Flowchart for platoon control strategy.</p>
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<p>The flow–density diagram.</p>
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<p>Comparison of road capacity.</p>
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<p>Spatiotemporal trajectory diagrams for the basic scenario.</p>
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<p>Spatiotemporal trajectory diagrams for the platoon control scenario.</p>
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<p>Spatiotemporal trajectory diagrams for the hybrid control framework scenario.</p>
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<p>Distributions of TTC, CIF, and DRAC under various MPRs of CAVs.</p>
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<p>Distributions of TTC, CIF, and DRAC under various MPRs of CAVs.</p>
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<p>Distributions of TET under various MPRs and traffic flow levels.</p>
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19 pages, 9213 KiB  
Article
Direct Ink Writing and Photocrosslinking of Hydroxypropyl Cellulose into Stable 3D Parts Using Methacrylation and Blending
by Mehmet-Talha Yapa, Gopakumar Sivasankarapillai, Jacques Lalevée and Marie-Pierre Laborie
Polymers 2025, 17(3), 278; https://doi.org/10.3390/polym17030278 - 22 Jan 2025
Abstract
Two 50% solid content solutions of methacrylated hydroxypropyl cellulose (MAHPC) with respective substitution degrees of 1.85 ± 0.04 (L_MAHPC) and 2.64 ± 0.04 (H_MAHPC) were screened for rheological properties, photocrosslinking kinetics and printability in relevance to direct ink writing (DIW). Photo-rheological and printability [...] Read more.
Two 50% solid content solutions of methacrylated hydroxypropyl cellulose (MAHPC) with respective substitution degrees of 1.85 ± 0.04 (L_MAHPC) and 2.64 ± 0.04 (H_MAHPC) were screened for rheological properties, photocrosslinking kinetics and printability in relevance to direct ink writing (DIW). Photo-rheological and printability studies reveal that the rheological properties of both MAHPC inks are better suited for DIW than those of hydroxypropyl cellulose (HPC) inks. Namely, methacrylate grafting improves shear dynamic moduli at low strain but also shear thinning and shear recovery. Both inks completely cure within 30 s upon shining UV light. Photocrosslinking is found to follow the phenomenological autocatalytic Sestak–Berggren kinetic model. However, prolonged exposure to UV light past full cure upon DIW leads to part fracture. The narrow UV-cure time window consequently precludes the production of multilayer parts using UV-assisted DIW for these neat MAHPC inks. In contrast, when blending MAHPC with HPC, an optimal balance between curing kinetics and DIW conditions is achieved, and stable, high-fidelity 150-layered parts are produced. Altogether this research highlights the need to design the content of photocrosslinkable moieties of cellulose derivatives to photoprint high fidelity and stable 3D parts from HPC inks. Full article
(This article belongs to the Section Biobased and Biodegradable Polymers)
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Graphical abstract

Graphical abstract
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<p>FTIR spectra of HPC and methacrylated HPCs.</p>
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<p><sup>1</sup>H-NMR spectra of HPC and methacrylated HPCs.</p>
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<p>Evolution of dynamic shear moduli of HPC and MAHPC inks upon exposure to UV light in a small oscillatory rheometer.</p>
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<p>Rate of mechanical cure as a function of mechanical conversion during photocrosslinking of MAHPC inks.</p>
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<p>Initial 3D-printability assessment of HPC and MAHPC inks a (<b>a</b>,<b>d</b>): HPC, (<b>b</b>,<b>e</b>): L_MAHPC, (<b>c</b>,<b>f</b>): H_MAHPC; Top: fiber/droplet formation essay, Bottom: layer stacking/merging essay.</p>
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<p>Shear-rate dependency of shear viscosity of HPC and MAHPC inks.</p>
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<p>Shear storage and loss moduli of HPC and MAHPC inks from amplitude sweeps conducted at 10 rad/s (τ<sub>y</sub>, yield point; τ<sub>f</sub>, flow point).</p>
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<p>Shear storage and loss moduli of HPC and MAHPC inks during a 3-segments recovery test where segments 1 and 3 operate within the LVE at 0.01% and segment 2 operates within the flow strain region at 500%.</p>
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<p>Reaction scheme for methacrylic grafting onto HPC.</p>
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19 pages, 4194 KiB  
Article
Optimized Methods to Quantify Tumor Treating Fields (TTFields)-Induced Permeabilization of Glioblastoma Cell Membranes
by Melisa Martinez-Paniagua, Sabbir Khan, Nikita W. Henning, Sri Vaishnavi Konagalla and Chirag B. Patel
Methods Protoc. 2025, 8(1), 10; https://doi.org/10.3390/mps8010010 - 22 Jan 2025
Abstract
Glioblastoma (GBM) is a lethal primary brain cancer with a 5.6% five-year survival rate. Tumor treating fields (TTFields) are alternating low-intensity electric fields that have demonstrated a GBM patient survival benefit. We previously reported that 0.5–24 h of TTFields exposure resulted in an [...] Read more.
Glioblastoma (GBM) is a lethal primary brain cancer with a 5.6% five-year survival rate. Tumor treating fields (TTFields) are alternating low-intensity electric fields that have demonstrated a GBM patient survival benefit. We previously reported that 0.5–24 h of TTFields exposure resulted in an increased uptake of FITC-dextran fluorescent probes (4–20 kDa) in human GBM cells. However, this approach, in which a fluorescence plate-based detector is used to evaluate cells attached to glass coverslips, cannot distinguish FITC-dextran uptake in live vs. dead cells. The goal of the study was to report the optimization and validation of two independent methods to quantify human GBM cell membrane permeabilization induced by TTFields exposure. First, we optimized flow cytometry by measuring mean fluorescence intensity at 72 h for 4 kDa (TTFields 6726 ± 958.0 vs. no-TTFields 5093 ± 239.7, p = 0.016) and 20 kDa (7087 ± 1137 vs. 5055 ± 897.8, p = 0.031) probes. Second, we measured the ratio of lactate dehydrogenase (LDH) to cell viability (measured using the CellTiter-Glo [CTG] viability assay); the LDH/CTG ratio was higher under TTFields (1.47 ± 0.15) than no-TTFields (1.08 ± 0.08) conditions, p < 0.0001. The findings using these two independent methods reproducibly demonstrated their utility for time-dependent evaluations. We also showed that these methods can be used to relate the cell membrane-permeabilizing effects of the non-ionizing radiation of TTFields to that of an established cell membrane permeabilizer, the non-ionic detergent Triton-X-100. Evaluating carboplatin ± TTFields, the LDH/CTG ratio was significantly higher in the TTFields vs. no-TTFields condition at each carboplatin concentration (0–30 µM), p = 0.014. We successfully optimized and validated two cost-effective methods to reproducibly quantify TTFields-induced human GBM cancer cell membrane permeabilization. Full article
(This article belongs to the Section Molecular and Cellular Biology)
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Figure 1
<p>Overview of the experimental setup for the in vitro TTFields experiment and methods for readouts.</p>
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<p>Determination of fluorescence probe concentration for FC. (<b>A</b>) Representative FC plots display the gating strategy to identify FITC-dextran-positive cells. U87 cells were incubated with FITC-dextran 4 or 20 kDa for 1 h, and 10,000 live FITC-positive cell events were recorded using FC. (<b>B</b>) The line plots show the MFI of the various concentrations of the FITC-dextran 4 or 20 kDa probes. The optimal concentration for each probe was determined to be 0.72 mg/mL. (<b>C</b>) The representative FC count histograms of the FITC-dextran probes at different concentrations. FITC = fluorescein isothiocyanate; FS = forward scatter; PI = propidium iodide.</p>
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<p>TTFields exposure (24–72 h) significantly increased the uptake of different-sized FITC-dextran probes (4 and 20 kDa). (<b>A</b>) Representative plots of the MFI of FITC-dextran probe uptake after a 10 min Triton-X-100 exposure. MFI of the (<b>B</b>) 4 kDa and (<b>C</b>) 20 kDa FITC-dextran probe uptake after TTFields exposure for 24–72 h. Normalized ratio of the FITC-dextran MFI in the TTFields-to-no-TTFields conditions for the (<b>D</b>) 4 kDa and (<b>E</b>) 20 kDa probes demonstrates that TTFields increased cell membrane permeability for the smaller 4 kDa probe, but not the larger 20 kDa probe, after a 48 h exposure. Three independent experiments were performed, and a parametric (#) or non-parametric test (*) was applied for statistical analysis. # or * indicates <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, and ### <span class="html-italic">p</span> &lt; 0.001 compared to the no-TTFields or control (unexposed) groups.</p>
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<p>Optimized LDH release assay detected TTFields-induced human GBM cell membrane permeabilization in a time-dependent manner, with high sensitivity. (<b>A</b>–<b>C</b>) The sensitivity of the LDH assay was evaluated by optimizing the dilution of the cell culture media samples after a 10 min exposure of the cells to Triton-X-100 (0.2% <span class="html-italic">v</span>/<span class="html-italic">v</span>). Control (gray bar) represents the different dilutions of the media samples that were collected from the cells that had previously not been exposed to Triton-X-100: (<b>A</b>) 1:10, (<b>B</b>) 1:50, and (<b>C</b>) 1:100. The indigo bar in (<b>A</b>–<b>C</b>) represents the media samples that had been exposed to Triton-X-100 (0.2% <span class="html-italic">v</span>/<span class="html-italic">v</span>). The two purple bars in (<b>A</b>–<b>C</b>) show the luminescence generated by the recombinant LDH reference standard diluted to 3200 mUnit/mL (light purple) and 32 mUnit/mL (dark purple), for comparison purposes. (<b>D</b>) Cell viability by CTG assay, (<b>E</b>) LDH release, and (<b>F</b>) normalized LDH/CTG ratio for cells unexposed to Triton-X-100 (control, gray) and exposed to Triton-X-100 for 10 min (0.01–0.025%, shades of indigo). (<b>G</b>) Cell viability by CTG assay, (<b>H</b>) LDH release, and (<b>I</b>) normalized LDH/CTG ratio for cells exposed to no-TTFields (gray) or exposed to TTFields (salmon) for 24–72 h. Three independent experiments were performed, and either a parametric test (#) or a non-parametric test (*) was applied for statistical analysis. * indicates <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, and ### or *** <span class="html-italic">p</span> &lt; 0.001 compared to the no-TTFields or control (unexposed) group. CTG = CellTiter-Glo; rLDH = recombinant lactate dehydrogenase.</p>
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<p>Normalized LDH/CTG ratio detected the GBM cell membrane permeabilization induced by the combination of TTFields with carboplatin chemotherapy in the surviving cells. (<b>A</b>) Cell viability was measured by CTG assay, (<b>B</b>) LDH release, and (<b>C</b>) normalized LDH/CTG in U87 cells exposed to carboplatin (0–30 µM) ± TTFields for 72 h. Two independent experiments were performed, and either a parametric test (#) or a non-parametric test (*) was applied for statistical analysis. # or * indicates <span class="html-italic">p</span> &lt; 0.05, ## or ** <span class="html-italic">p</span> &lt; 0.01, and ### or ***<span class="html-italic">p</span> &lt; 0.001 compared to the no-TTFields or control (unexposed) group. CTG = CellTiter-Glo; LDH = lactate dehydrogenase; ns = not statistically significant.</p>
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<p>Graphical summary of the methods used in human GBM cells for evaluating TTFields-induced cell membrane permeabilization. Created with BioRender.com.</p>
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13 pages, 895 KiB  
Article
Use of Ozone for Disinfection of PHARMODUCT® Automatic System for Antineoplastic Compounding
by Vito Lovino, Antonio Riglietti, Anna Tolomeo, Giuseppe Capasso, Miriana Di Vittorio, Stefano Brattoli, Giuseppe Tesse, Vincenzo Dimiccoli, Marco Spartà and Luana Perioli
Pharmaceuticals 2025, 18(2), 140; https://doi.org/10.3390/ph18020140 - 22 Jan 2025
Viewed by 95
Abstract
Background: The purpose of this work was to demonstrate the ozone efficacy for disinfection of the PHARMODUCT® automatic dispensing system for antineoplastic preparation, as a guarantee of a higher grade of cleanliness. While the use of ozone gas disinfection is almost consolidated [...] Read more.
Background: The purpose of this work was to demonstrate the ozone efficacy for disinfection of the PHARMODUCT® automatic dispensing system for antineoplastic preparation, as a guarantee of a higher grade of cleanliness. While the use of ozone gas disinfection is almost consolidated in food and water treatment, there is a lack of scientific data in the pharmaceutical field. The scope of this study was to demonstrate the ozone efficacy for disinfection of the PHARMODUCT® automatic dispensing system, before starting the antineoplastic preparation, in order to ensure a high degree of cleanliness and, at the same time, to define a biodecontamination procedure that could also be translatable to other automated compounding systems on the market. Methods: Ozone efficacy was determined by calculating the difference (pre-exposure–post-exposure) in CFU counts on the plate. A group of four different ATCC-selected microbial strains were tested using two distinct cycles. The first one was evaluated with an ozone gas concentration of 40 ppm for 40 min; the second cycle increased the concentration to 60 ppm for the same duration. Results: Results showed that exposure to 40 ppm ozone gas led to a 4-log reduction of all tested ATCC strains. In contrast, exposure to 60 ppm ensured a 6-log reduction. Conclusions: The ozone disinfection process, applied to the PHARMODUCT® system, provides a superior grade of cleanliness compared to the manual disinfection procedure, thus offering insight beyond the current anti-inflammatory and analgesic application of ozone therapy in the medical field. Full article
(This article belongs to the Section Pharmaceutical Technology)
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Graphical abstract

Graphical abstract
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<p>The outside of the PHARMODUCT<sup>®</sup> automatic system built by Bioduct s.r.l. Firenze 50141, Italy, which is used for antineoplastic preparation.</p>
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<p>Layout of PHARMODUCT<sup>®</sup> main chamber with plate positions.</p>
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22 pages, 3141 KiB  
Article
Long-Term Exposure to Lambda-Cyhalothrin Reveals Novel Genes Potentially Involved in Aedes aegypti Insecticide Resistance
by Alejandro Mejía, Ana María Mejía-Jaramillo, Geysson Javier Fernandez, Yurany Granada, Carl Lowenberger and Omar Triana-Chávez
Insects 2025, 16(2), 106; https://doi.org/10.3390/insects16020106 - 21 Jan 2025
Viewed by 229
Abstract
Insecticide resistance in Aedes aegypti populations hinders vector control programs. Many studies have focused on the classical mechanisms, kdr mutations, and metabolic enzymes to understand the development of insecticide resistance. In this study, we subjected a strain of Ae. aegypti to selective pressure [...] Read more.
Insecticide resistance in Aedes aegypti populations hinders vector control programs. Many studies have focused on the classical mechanisms, kdr mutations, and metabolic enzymes to understand the development of insecticide resistance. In this study, we subjected a strain of Ae. aegypti to selective pressure for 13 consecutive generations to understand the development and extent of insecticide resistance. We delved into the transcriptomics of this pressured strain to gain insights into the molecular changes underlying insecticide resistance in Ae. aegypti. Our data suggest mosquito resistance is influenced by additional mechanisms that are difficult to explain using only classical mechanisms. The response by mosquitoes varies depending on the exposure time. Initially, when mosquitoes are in contact with insecticides, they modulate the expression of metabolic enzymes and gain some point mutations in the sodium channel genes. After long-term exposure, the mosquitoes respond to insecticides by expressing different proteins involved in the cuticle, energetic metabolism, and synthesis of proteases. We propose a model that includes these novel mechanisms found after prolonged insecticide exposure, which work in conjunction with established mechanisms (kdr and metabolic resistance) but have a different timeline in terms of expression and appearance. Full article
(This article belongs to the Section Medical and Livestock Entomology)
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Figure 1

Figure 1
<p>Experimental design overview. The Acacías population AF-0 was collected as stated by Granada et al. [<a href="#B16-insects-16-00106" class="html-bibr">16</a>], and following WHO protocols, the pressure was performed at the larval stage. Three biological replicates were performed to extract RNA and proceed with sequencing. Vector Base, Gene Ontology categories, and DAVID were consulted to obtain functional annotation of the DEGs (Differentially Expressed Genes).</p>
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<p><span class="html-italic">Kdr</span> mutation frequencies for mutations (<b>A</b>) V410L, (<b>B</b>) V1016I, and (<b>C</b>) F1534C in unpressured (AF13WP) and pressured (AF13P) populations. Allelic frequencies of all the populations in this study. Blue represents susceptible alleles, and purple represents resistant alleles. The F7 pressured (AF7P) and unpressured (AF7WP) allelic frequencies data reported by Granada et al. [<a href="#B16-insects-16-00106" class="html-bibr">16</a>] are also shown.</p>
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<p>Enzyme activity for <span class="html-italic">Ae. aegypti</span> populations. (<b>A</b>) Acetylcholinesterase (AChE), (<b>B</b>) mixed function oxidase (MFO), (<b>C</b>) α-esterase (α-EST), (<b>D</b>) β-esterase (β-EST), and (<b>E</b>) Glutathione-S-transferase (GST). Forty mosquitoes were used for each assay. Boxes and whiskers correspond to mean and percentiles 5–95. Asterisks correspond to significant differences (Kruskal Wallis, *** = <span class="html-italic">p</span> &lt; 0.0001).</p>
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<p>(<b>A</b>) Volcano Plot of False Discovery Rate vs. log 2-fold Change. The TOP 4 most downregulated and upregulated genes and their corresponding ID or annotation description are in the upper left and right corners. The same genes are in boxes in the lower left and right corners. Blue dots represent genes that surpassed only the FDR threshold of 0.05 but remained lower than log<sub>2</sub>FC of 1. Green dots are statistically non-significant (FDR &gt; 0.05) but exceeded the log<sub>2</sub>FC threshold of 1. Meanwhile, the red dots correspond to statistically significant DEGs (FDR &lt; 0.05) and have log<sub>2</sub>FC &gt; 1. Grey dots have no statistical significance (NS) nor representative log<sub>2</sub>FC. FDR is represented as a negative log-transformed <span class="html-italic">p</span>-value (−Log<sub>10</sub>P). (<b>B</b>) PCA of intrasample variation. Each sample’s normalized and scaled counts were used to plot the Principal Component Analysis (PCA). Letters A, B, and C represent replicates for AF13P and AF13WP.</p>
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<p>GO terms of upregulated (<b>A</b>) and downregulated (<b>B</b>) genes. The size of the bubbles corresponds to gene count in that specific term, while the color represents the associated <span class="html-italic">p</span>-value. Enrichment score corresponds to the number of genes with that term divided by the background.</p>
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<p>According to DAVID clustering and manual search, violin plots show specific gene regulation in 9 groups. The gray color corresponds to the area of no significant regulation. The dotted line determines the threshold of &gt;1 and &lt;−1 log<sub>2</sub>FC significant upregulated and downregulated genes. Blue dots represent specific downregulated genes, and purple dots indicate upregulated genes.</p>
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<p>The model proposed for the response to Lambda-cyhalothrin insecticide in <span class="html-italic">Aedes aegypti</span>. Early and late mechanisms are indicated on the left and right sides. Classical mechanisms (“early”) correspond to <span class="html-italic">kdr</span> and metabolic resistance in response to insecticides. Unexplored mechanisms (“late”) correspond to mechanisms found in response to prolonged insecticide exposure in AF13P.</p>
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22 pages, 3912 KiB  
Article
Complex Actions of FKBP12 on RyR1 Ion Channel Activity Consistent with Negative Co-Operativity in FKBP12 Binding to the RyR1 Tetramer
by Spencer J. Richardson, Chris G. Thekkedam, Marco G. Casarotto, Nicole A. Beard and Angela F. Dulhunty
Cells 2025, 14(3), 157; https://doi.org/10.3390/cells14030157 - 21 Jan 2025
Viewed by 264
Abstract
The association of the 12 KDa FK506 binding protein (FKBP12) with ryanodine receptor type 1 (RyR1) in skeletal muscle is thought to suppress RyR1 channel opening and contribute to healthy muscle function. The strongest evidence for this role is increased RyR1 channel activity [...] Read more.
The association of the 12 KDa FK506 binding protein (FKBP12) with ryanodine receptor type 1 (RyR1) in skeletal muscle is thought to suppress RyR1 channel opening and contribute to healthy muscle function. The strongest evidence for this role is increased RyR1 channel activity following FKBP12 dissociation. However, the corollary that channel activity will decrease when FKBP12 is added back to FKBP12-depleted RyR1 is not well established, and when reported, the time- and concentration-dependence of inhibition vary over orders of magnitude. Here, we address this problem with an investigation of the molecular mechanisms of the FKBP12 regulation of RyR1. Muscle processing to obtain sarcoplasmic reticulum (SR) vesicle preparations enriched in RyR1 resulted in substantial FKBP12 dissociation from RyR1, indicating low-affinity binding. Conversely, high-affinity binding was indicated by some FKBP12 remaining bound to RyR1 after solubilization. We report, for the first time, an increase in the activity of FKBP12-depleted channels after the addition of exogenous FKBP12 (5 nM to 5 µM), followed by a reduction in activity consistent with inhibition after 20–30 min exposure to higher [FKBP12]s. Both the increase and later decline in activity were time- and concentration-dependent. The results suggest a high-affinity activation when FKBP12 binding sites on the RyR1 tetramer are partially occupied by FKBP12 and lower affinity inhibition as more RyR1 monomers become occupied. These novel results imply negative cooperativity in FKBP12 binding to RyR1 and a dynamic role for FKBP12/RyR1 interactions in intact muscle fibers. Full article
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Figure 1
<p>FKBP12 dissociation from RyR1 during SR processing. (<b>A</b>) Sequential centrifugation and sucrose gradient fractionation in isolating SR vesicles enriched in RyR1 channels and solubilized RyR1. Relevant solute fractions are labeled S1 and S5, the relevant pellet is labeled P2, and sucrose gradient bands are labeled B1 to B4. (<b>B</b>–<b>H</b>) Co-IP of RyR1 and FKBP12 in SR preparations. Co-IP samples were loaded at protein concentrations of approximately 3, 6, and 9 μg. (<b>B</b>) RyR1 and FKBP12 in homogenate (H) and B4 vesicles (<span class="html-italic">n</span> = 5). (<b>C</b>) RyR1 and FKBP12 in P2 and B4 vesicles (<span class="html-italic">n</span> = 6). (<b>D</b>) RyR1 and FKBP12 in control-incubated homogenate and homogenate incubated with GST-cleaved FKBP12 (cFKBP12) (<span class="html-italic">n</span> = 3). (<b>E</b>) RyR1 and FKBP12 in B4 and solubilized B4 (<span class="html-italic">n</span> = 5). (<b>F</b>) RyR1, GST-FKBP12, and FKBP12 in incubated B4 without GST-FKBP12 (control) and B4 incubated with GST-FKBP12 (<span class="html-italic">n</span> = 4). (<b>G</b>) Supernatants following Co-IP control-incubated B4 and B4 incubated with GST-FKBP12 (<span class="html-italic">n</span> = 5). (<b>H</b>) B4 and B4 incubated with GST-cleaved FKBP12 (cFKBP12) (<span class="html-italic">n</span> = 3). (<b>B</b>–<b>H</b>) The images in each figure show immune-stained bands from the same SDS-PAGE gel. The upper image in each panel shows the RyR1 band, and the lower image shows the FKBP12 band. An additional band is shown for GST-GKBP12 in (<b>F</b>,<b>G</b>). All vertically aligned images in one panel were obtained from the same lane. MW markers are labeled in (<b>B</b>,<b>F</b>), and corresponding arrows indicating marker positions are shown in all panels. Unless otherwise stated, the graphs in each figure show the average relative FKBP12/RyR1 ratios. The ratios were first calculated for RyR and FKBP bands in the same lanes, and then, the ratio for the right-hand lane was expressed relative to ratios for the left-hand “control” lane in the same blot. The average values are indicated by broad vertical bars, and the s.e.m. is indicated by the vertical capped lines. The <span class="html-italic">n</span> values refer to the number of individual experiments. The total FKBP12 density in (<b>F</b>) was calculated as (FKBP12 + GST-FKBP12/2). The asterisks * in (<b>B</b>–<b>H</b>) indicate significant differences from the control, left hand bars.</p>
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<p>S107 prevents FKBP12’s dissociation from RyR1 during processing. Results for the Co-IP of RyR1 and FKBP12 in SR preparations in the following preparation in the presence of 20 µM of S107. Co-IP samples were loaded at protein concentrations of approximately 3, 6, and 9 μg. (<b>A</b>) RyR1 and FKBP12 in homogenate (H) and B4 vesicles in the presence of S107, as labeled. (<b>B</b>) RyR1 and FKBP12 in P2 and B4 vesicles in the presence of S107, as labeled. The images in each panel show bands from the same SDS-PAGE gel, and images in (<b>A</b>,<b>B</b>) are from the same gel as the images shown in <a href="#cells-14-00157-f001" class="html-fig">Figure 1</a>B,C, respectively. The upper image in each panel shows the RyR1 band, and the lower image shows the FKBP12 band, with vertically aligned images obtained from the same lane. Positions of MW markers are provided in (<b>A</b>) with corresponding arrows indicating marker positions in (<b>A,B</b>). The graphs (<b>A</b>) show the average relative FKBP12/RyR1 in the homogenate (<b>left</b>) and in B4 (<b>right</b>) processed with S107 (<span class="html-italic">n</span> = 5). The FKBP12/RyR1 ratios were first calculated for the RyR1 and FKBP12 bands in the same lanes; then, the H + S107 ratio was expressed relative to the H ratio (<b>left</b>), and the B4 + S107 ratio was expressed relative to the B4 ratio (<b>right</b>). The graphs in (<b>B</b>) show the average relative FKBP12/RyR1 ratio in P2 (<b>left</b>) and B4 (<b>right</b>) processed with S107 (<span class="html-italic">n</span> = 6). The FKBP12/RyR1 ratios were first calculated for the RyR1 and FKBP12 bands in the same lanes, and then, the ratio for P2 + S107 was expressed relative to the ratio in P2 alone, and the ratio for B4 + S107 was expressed relative to the ratio for B4 alone. The average values are indicated by broad vertical bars, and the s.e.m. is indicated by the vertical capped lines. The <span class="html-italic">n</span> values refer to the number of individual experiments. (<b>C</b>) A summary of results obtained with vesicle processing in the absence of S107 (black) and the presence (cyan) of S107. The asterisk * indicates a significant difference from the control, and the @ symbol indicates a significant difference from the S107 data for the P2 data.</p>
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<p>Biphasic FKBP12-induced changes in the activity of native RyR1 channels from B3 and B4 sucrose gradient fractions. RyR1 channel activity recorded in artificial lipid bilayers. (<b>A</b>) Current recordings obtained from native RyR1 channels during typical experiments over 4 to 8 min of control recording before FKBP12 was added to the cytoplasmic (cis) solution containing different [Ca<sup>2+</sup>] and [ATP] specified above the records. Upper “control” recordings were obtained before FKBP12 addition, and the lower recordings at the times below each record. The [FKBP12]s are provided at the top of each panel. Currents were at +40 mV, with upward deflections indicating channel opening from the baseline (indicated by the arrow on the left-hand side of each row) to the maximum open channel level (broken lines in each record). (<b>i</b>) 10 µM cis Ca<sup>2+</sup>: No change in activity after 12 min exposure to 5 nM FKBP12. (<b>ii</b>) 1 µM cis Ca<sup>2+</sup>: increased activity after 23 min with 5 nM FKBP12. (<b>iii</b>) 1 µM cis Ca<sup>2+</sup>: increased activity after 5 min with 0.2 µM FKBP12. (<b>iv</b>) 1 µM cis Ca<sup>2+</sup>: increased activity after 9 min with 1.0 µM FKBP12. (<b>v</b>) 1 µM cis Ca<sup>2+</sup>: increased activity after 5 min with 5.0 µM FKBP12. (<b>B</b>–<b>D</b>) Analysis of channel activity as a function of time and [FKBP12]. <span class="html-italic">P</span><sub>o</sub> was measured from single-channel recordings with 1 µM cis Ca<sup>2+</sup> and <span class="html-italic">I</span>′<span class="html-italic">F</span> for multiple channel recordings with 10 µM cis Ca<sup>2+</sup> and 10 µM cis Ca<sup>2+</sup> plus 2 mM ATP in the cytoplasmic solutions. Data are plotted as a function of time after FKBP12 addition to the cytoplasmic solution. (<b>B</b>) 1–5 nM FKBP12 with 1 µM cis Ca<sup>2+</sup> (royal blue symbols, <span class="html-italic">n</span> = 12), 10 µM cis Ca<sup>2+</sup> (black symbols, <span class="html-italic">n</span> = 8), and 10 µM cis Ca<sup>2+</sup> plus 2 mM ATP (cyan symbols, <span class="html-italic">n</span> = 12). (<b>C</b>) 0.2 µM FKBP12 (<span class="html-italic">n</span> = 6). (<b>D</b>) 1 µM FKBP12 (<span class="html-italic">n</span> = 10) and <span class="html-italic">n</span> = 2 for 5 µM FKBP12). The data for the one channel recorded with 5 µM FKBP12 are included because they provide additional evidence confirming the biphasic trends seen with 1 µM FKBP12. (<b>B</b>(<b>i</b>)–<b>D</b>(<b>i</b>)) Graphs showing individual measurements of relative <span class="html-italic">P</span><sub>o</sub> and relative <span class="html-italic">I</span>′<span class="html-italic">F</span> at +40 mV (filled circles) and at −40 mV (open circles) for all channels contributing to the average data in (<b>B</b>(<b>ii</b>)–<b>D</b>(<b>ii</b>)). (<b>B</b>(<b>ii</b>)–<b>D</b>(<b>ii</b>)) Graphs showing average combined relative <span class="html-italic">P</span><sub>o</sub> and relative <span class="html-italic">I</span>′<span class="html-italic">F</span> in (<b>Bii</b>) or average relative <span class="html-italic">P</span><sub>o</sub> only in (<b>C</b>(<b>ii</b>),<b>D</b>(<b>ii</b>)). All data in (<b>C</b>,<b>D</b>) were obtained from single-channel recordings with 1 µM cis Ca<sup>2+</sup>. (<b>B</b>(<b>iii</b>)–<b>D</b>(<b>iii</b>)) Graphs showing average <span class="html-italic">P</span><sub>o</sub> and average <span class="html-italic">I</span>′<span class="html-italic">F</span> in (<b>B</b>(<b>iii</b>)) or average <span class="html-italic">P</span><sub>o</sub> in (<b>C</b>(<b>iii</b>),<b>D</b>(<b>iii</b>)). In columns (<b>ii</b>,<b>iii</b>), filled symbols indicate the average values, with the s.e.m. indicated by vertical capped bars. (<b>E</b>) Data for each [FKBP12] shown in (<b>B</b>–<b>D</b>) have been rearranged into three time groups: 1–5 min, left bar; 7–10 min, middle bar; and 11–20 min, right bar. For 1–10 nM FKBP12, 1–5 min (<span class="html-italic">n</span> = 20); 7–10 min (<span class="html-italic">n</span> = 8); 11–20 min (<span class="html-italic">n</span> = 4). For 0.2 µM FKBP12, 1–5 min (<span class="html-italic">n</span> = 12); 7–10 min (<span class="html-italic">n</span> = 6); 11–20 min (<span class="html-italic">n</span> = 6). For 1.0 µM FKBP12, 1–5 min (<span class="html-italic">n</span> = 12); 7–10 min (<span class="html-italic">n</span> = 7); 11–20 min (<span class="html-italic">n</span> = 8). Blue asterisks * in (<b>B</b>–<b>E</b>) indicate significant differences from the control after the addition of FKBP12. The red asterisks (*) in (E) indicate significant differences from the same time period in 1–5 nM FKBP12. The red @ symbol in (<b>E</b>) indicates significant differences from 7 to 10 min in 1–5 nM FKBP12.</p>
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<p>Changes in native RyR1 single-channel gating parameters during exposure to FKBP12. Gating parameters, mean open time (<span class="html-italic">T<sub>o</sub></span>, left column), mean closed time (<span class="html-italic">T<sub>c</sub></span>, middle column), and event frequency (<span class="html-italic">F<sub>o</sub></span>, right column) were measured from the single-channel recording at the same time as the <span class="html-italic">P<sub>o</sub></span> values in <a href="#cells-14-00157-f003" class="html-fig">Figure 3</a>. Refer to <a href="#cells-14-00157-f003" class="html-fig">Figure 3</a> for general details that are not repeated here and for the number of experiments. Average data are plotted as a function of time and [FKBP12]. (<b>A</b>) 1–5 nM FKBP12; (<b>B</b>) 0.2 µM FKBP12; (<b>C</b>) 1 and 5 µM FKBP12. The data for the one channel recorded with 5 µM FKBP12 are included because they provide additional support for the trends seen with 1 µM FKBP12. Filled symbols indicate the average values, with the s.e.m. indicated by vertical capped bars. The broken red lines surround data sets that show trends in the gating parameter that are most clearly associated with the rapid onset of activation (<span class="html-italic">T<sub>o</sub></span> in (<b>A</b>)). Blue asterisks * in indicate significant differences from the control after the addition of FKBP12.</p>
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<p>Effects of FBP12 on partially purified solubilized RyR1 channels. Ion channel activity recorded in artificial lipid bilayers. Current recordings obtained from solubilized RyR1 channels during typical experiments during 4 to 8 min control recording before FKBP12 was added to the cytoplasmic solution containing 1 µM free Ca<sup>2+</sup> and for periods of ~20–38 min in the presence of FKBP12. (<b>A</b>–<b>D</b>) Column (<b>i</b>) shows examples of channel currents recorded before (upper record) and at the indicated times after FKBP12 addition (lower records). Currents are shown at +40 mV, with upward deflections indicating channel opening. In each recording, the solid red line marks the baseline (zero current level), and the broken red line marks the maximum open level for a single channel. The open level for a second full conductance opening is not specifically marked, as it is seldom clearly seen and cannot be separated from multiple sub-conductance openings. Column (<b>ii</b>): Plots of individual measurements of relative <span class="html-italic">I</span>′<span class="html-italic">F</span> at +40 mV (filled circles) and at −40 mV (open circles). Column (<b>iii</b>): average relative <span class="html-italic">I</span>′<span class="html-italic">F</span> (including independent values at +40 and −40 mV). Column (<b>iv</b>): Average <span class="html-italic">I</span>′<span class="html-italic">F</span>, shown to illustrate the approximate open probability of the solubilized channels (as described in the text). (<b>A</b>) In the initial 2 experiments, FKBP12 was added first at 1 nM and then increased to 5 nM (sky blue symbols). In the following 4 experiments, 5 nM FKBP12 was added first, and then, the concentration increased to 10 nM (royal blue symbols). The data from the two protocols were combined in the average effect of these low concentrations (<span class="html-italic">n</span> = 12). (<b>B</b>) Exposure to 0.2 µM FKBP12 (<span class="html-italic">n</span> = 8). (<b>C</b>) Exposure to 1.0 µM FKBP12 (<span class="html-italic">n</span> = 14). (<b>D</b>) Exposure to 5.0 µM FKBP12 (n = 16). (<b>E</b>) The data in (<b>A</b>–<b>D</b>) have been regrouped into three periods: For 1–10 nM FKBP12, 1–5 min (<span class="html-italic">n</span> = 24); 10–20 min (<span class="html-italic">n</span> = 23); 22–30 min (<span class="html-italic">n</span> = 27). For 0.2 µM FKBP12, 1–5 min (<span class="html-italic">n</span> = 16); 9–15 min (<span class="html-italic">n</span> = 16); 17–20 min (<span class="html-italic">n</span> = 16). For 1.0 µM FKBP12, 1–5 min (<span class="html-italic">n</span> = 28); 9–15 min (<span class="html-italic">n</span> = 28); 16–21 min (<span class="html-italic">n</span> = 20). For 5.0 µM FKBP12, 1–5 min (<span class="html-italic">n</span> = 32); 11–17 min (<span class="html-italic">n</span> = 28); 18–30 min (<span class="html-italic">n</span> = 18). The <span class="html-italic">n</span> values refer to the number of observations within each group. Blue asterisks (<b>*</b>) in (<b>A</b>–<b>E</b>) indicate significant differences from the control after the addition of FKBP12. The red asterisks (*) in (<b>E</b>) indicate significant differences from the same time period in 1–5 nM FKBP12. For average data in (<b>A</b>,<b>D</b>), symbols show average values, and the s.e.m. is indicated by the vertical bars.</p>
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<p>(<b>A</b>,<b>B</b>) FKBP12 inhibition of RyR1 is enhanced by S107. Ion channel activity recorded from native RyR1 channels (<b>A</b>(<b>i</b>)) and from solubilized RyR1 channels (<b>B</b>(<b>i</b>)), before FKBP12 addition to the cytoplasmic solution containing 1 µM free Ca<sup>2+</sup> and then at various times over ~20–38 min exposures to 0.2 µM FKBP12 alone or 0.2 µM FKBP12 with 20 µM S107 added 5–10 min after FKBP12. Currents are shown at +40 mV, with upward deflections indicating channel opening. Relative <span class="html-italic">P<sub>o</sub></span> for native RyR1 channels and <span class="html-italic">I</span>′<span class="html-italic">F</span> for solubilized RyR1 exposed to 0.2 µM FKBP12 in the absence of S107 in <a href="#cells-14-00157-f003" class="html-fig">Figure 3</a> and <a href="#cells-14-00157-f005" class="html-fig">Figure 5</a> are included in (<b>A</b>(<b>ii</b>),<b>B</b>(<b>ii</b>)), respectively, for comparison with the S107 data sets. Measurements of relative <span class="html-italic">P<sub>o</sub></span> or relative <span class="html-italic">I</span>′<span class="html-italic">F</span> at +40 mV and −40 mV are included in the average relative <span class="html-italic">P<sub>o</sub></span> and relative <span class="html-italic">I</span>′<span class="html-italic">F</span>, as described in the legends of <a href="#cells-14-00157-f003" class="html-fig">Figure 3</a> and <a href="#cells-14-00157-f005" class="html-fig">Figure 5</a>. (<b>A</b>(<b>ii</b>),<b>B</b>(<b>ii</b>)) Individual measurements of relative <span class="html-italic">P<sub>o</sub></span> or relative <span class="html-italic">I</span>′<span class="html-italic">F</span> at +40 mV (filled circles) and −40 mV (open circles) with 0.2 µM FKBP12 alone (royal blue symbols) or with 0.2 µM FKBP12 + S107 (cyan symbols). (<b>A</b>(<b>iii</b>),(<b>Biii</b>)) Graphs of average relative <span class="html-italic">P<sub>o</sub></span> for native RyR1 channels (<span class="html-italic">n</span> = 6) and average <span class="html-italic">I</span>′<span class="html-italic">F</span> for solubilized RyR1 channels (<span class="html-italic">n</span> = 4), respectively, with 0.2 µM FKBP12 alone (royal blue symbols) or with 0.2 µM FKBP12 + S107 (cyan symbols). The red arrows in the graphs indicate approximate times of S107 addition to the cytoplasmic solution. (<b>C</b>,<b>D</b>) Effects of FKBP12 on current crossing the bilayer membrane, revealed in parameters used to calculate <span class="html-italic">I</span>′<span class="html-italic">F</span> for solubilized RyR1. The <span class="html-italic">Imean</span> data in (<b>C</b>) and <span class="html-italic">Imax</span> data in (<b>D</b>) were used to calculate <span class="html-italic">I</span>′<span class="html-italic">F</span> in <a href="#cells-14-00157-f005" class="html-fig">Figure 5</a>. Thus the number of experiments are the same as in <a href="#cells-14-00157-f005" class="html-fig">Figure 5</a> and number of observations are provided in the legend to <a href="#cells-14-00157-f005" class="html-fig">Figure 5</a>. The upper graphs in each panel in (<b>C</b>,<b>D</b>) show individual <span class="html-italic">Imean</span> and <span class="html-italic">Imax</span> values, respectively, at +40 mV (filled circles) and −40 mV (open circles), while the lower graphs show average <span class="html-italic">Imean</span> and average <span class="html-italic">Imax</span>. (<b>C</b>(<b>i</b>),<b>D</b>(<b>i</b>)) 1–10 nM FKBP12: Sky blue symbols are from channels exposed to 1 nM and then 5 nM FKBP12, and royal blue symbols are from channels exposed to 5 nM and then 10 nM FKBP12; (<b>C</b>(<b>ii</b>),<b>D</b>(<b>ii</b>)) 0.2 µM FKBP12; (<b>C</b>(<b>iii</b>),<b>D</b>(<b>iii</b>)); 1.0 µM FKBP12; (<b>C</b>(<b>iv</b>),<b>D</b>(<b>iv</b>)) 5.0 µM FKBP12. The data from the 1–5 nM and 5–10 nM groups were combined to calculate the average effect of these low concentrations. For average data in (<b>A</b>–<b>D</b>), symbols indicate average values, and the s.e.m. is indicated by the vertical bars. The <span class="html-italic">n</span> values refer to the number of observations. The blue asterisks * indicate a significant difference from the control and the red asterisk * indicates a significant difference between the indicated data with S107 and without S107.</p>
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<p>Normalization highlights the dominance of increases in <span class="html-italic">Imean</span> in the effects of FKBP12 on solubilized RyR1 channel activity and <span class="html-italic">I</span>′<span class="html-italic">F</span>. The relative <span class="html-italic">Imean</span> and <span class="html-italic">Imax</span> were calculated for the <span class="html-italic">Imean</span> and <span class="html-italic">Imax</span> data in <a href="#cells-14-00157-f005" class="html-fig">Figure 5</a> and <a href="#cells-14-00157-f006" class="html-fig">Figure 6</a>. Both <span class="html-italic">Imean</span> and <span class="html-italic">Imax</span> in the presence of FKBP12 were normalized to their control value for each individual bilayer before obtaining the average relative values. Details of the experiments, including the number of observations, are the same as those in the legend in <a href="#cells-14-00157-f005" class="html-fig">Figure 5</a>. In (<b>A</b>–<b>E</b>), panel (<b>i</b>) shows the relative <span class="html-italic">Imax</span> in individual bilayers at +40 mV (filled circles) and −40 mV (open circles). Panel (<b>ii</b>) shows the average relative <span class="html-italic">Imax</span>. Panel (<b>iii</b>) shows relative <span class="html-italic">Imean</span> from individual bilayers at +40 mV (filled circles) and −40 mV (open circles). Panel (<b>iv</b>) shows the average relative <span class="html-italic">Imean</span>. For average data <b>i</b>n (<b>ii</b>,<b>iv</b>), symbols show average relative values, and the s.e.m. is indicated by the vertical capped bars. Data are shown in (<b>A</b>) for 1–10 nM FKBP12, in (<b>B</b>) for 0.2 µM FKBP12, in (<b>C</b>) for 1.0 µM FKBP12, and in (<b>D</b>) for 5.0 µM FKBP12, and in (<b>E</b>), data for 0.2 µM FKBP12 are repeated (filled royal blue circles) for comparison with 0.2 µM FKBP12 + S107 (cyan circles). The blue asterisks * indicate a significant difference between data with FKBP12 alone and control data. The cyan asterisks * indicate a significant difference between data with FKBP12 plus S107 and control data for those experiments and the red asterisk * indicates a significant difference between the indicated data with S107 and without S107.</p>
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<p>The negative co-operativity model suggests that changes in subunit occupation by FKBP12 within each tetramer might explain the changes in RyR1 channel activity associated with exogenous FKBP12 binding. (<b>A</b>,<b>B</b>) illustrate patterns of FKBP12 occupation of RyR1 subunits in RyR1 tetramers from B4 vesicles (<b>A</b>) and solubilized RyR1 (<b>B</b>). The control horizontal panel for B4 vesicles shows examples of four control RyR1 tetramers before adding exogenous FKBP12.From left to right, 1 tertamer with no FkBP12 bound, 2 tetramers with 1 FKBP12 bound, and 1 tetramer with 2 FKBP12 molecules bound, approximating the mean of 1.25 (Results). The control horizontal panel for four solubilized RyR1 tetramers is shown with 3 tetramers having 0 FKBP12 molecules bound, and 1 RyR1 tetramer with 1 FKBP bound. This is an overestimate of the 0.19 tetramers with FKBP12 bound (Discussion) but is shown to indicate that a small but finite number of solubilized RyR1 channels contain 1 subunit with FKBP12 bound. The remaining panels show increasing FKBP12 occupation with increasing time after exogenous FKBP12 addition and increasing [FKBP12]. The changes in occupation are suggested to explain the biphasic effects of FKBP12 on RyR1 channel activity presented in <a href="#cells-14-00157-f003" class="html-fig">Figure 3</a>, <a href="#cells-14-00157-f004" class="html-fig">Figure 4</a>, <a href="#cells-14-00157-f005" class="html-fig">Figure 5</a>, <a href="#cells-14-00157-f006" class="html-fig">Figure 6</a> and <a href="#cells-14-00157-f007" class="html-fig">Figure 7</a> and outlined in the Discussion. Note that the activated RyR1 tetramers are illustrated as having a larger cross-sectional profile as seen in the corona of activated RyRs, in contrast to the inhibited tetramers, which are depicted as smaller, representing the compact corona in a more closed conformation.</p>
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17 pages, 248 KiB  
Review
Sustainable Architecture and Human Health: A Case for Effective Circadian Daylighting Metrics
by Bhaswati Mukherjee and Mohamed Boubekri
Buildings 2025, 15(3), 315; https://doi.org/10.3390/buildings15030315 - 21 Jan 2025
Viewed by 305
Abstract
The development of the fluorescent lamp and the air-conditioning system resulted in buildings being lit inexpensively without having to rely on daylighting to save energy, as was the case during the incandescent lamp era. Consequently, architects were able to design buildings with deep [...] Read more.
The development of the fluorescent lamp and the air-conditioning system resulted in buildings being lit inexpensively without having to rely on daylighting to save energy, as was the case during the incandescent lamp era. Consequently, architects were able to design buildings with deep floor plates for maximum occupancy, placing workstations far away from windows since daylighting was no longer a necessity. Floor-to-ceiling heights became lower to minimize the inhabitable volumes that needed to be cooled or heated. With the rising costs of land in some major American cities such as New York City and Chicago at the beginning of the twentieth century, developers sought to optimize their investments by erecting tall structures, giving rise to densely inhabited city centers with massive street canyons that limit sunlight access in the streets. Today, there is growing awareness in terms of the impact of the built environment on people’s health especially in terms of the health benefits of natural light. The fact that buildings, through their shapes and envelope, filter a large amount of daylight, which may impact building occupants’ health and well-being, should cause architects and building developers to take this issue seriously. The amount and quality of light we receive daily impacts many of our bodily functions and consequently several aspects of our health and well-being. The human circadian rhythm is entrained by intrinsically photosensitive retinal ganglion cells (ipRGCs) in our eyes that are responsible for non-visual responses due to the presence of a short-wavelength sensitive pigment called melanopsin. The entrainment of the circadian rhythm depends on several factors such as the intensity, wavelength, timing, and duration of light exposure. Recently, this field of research has gained popularity, and several researchers have tried to create metrics to quantify photopic light, which is the standard way of measuring visual light, into a measure of circadian effective lighting. This paper discusses the relationship between different parameters of daylighting and their non-visual effects on the human body. It also summarizes the existing metrics of daylighting, especially those focusing on its effects on the human circadian rhythm and its shortcomings. Finally, it discusses areas of future research that can address these shortcomings and potentially pave the way for a universally acceptable standardized metric. Full article
(This article belongs to the Section Architectural Design, Urban Science, and Real Estate)
20 pages, 2734 KiB  
Review
Form and Function: The Factors That Influence the Efficacy of Nanomaterials for Gene Transfer to Plants
by Zhila Osmani and Marianna Kulka
Molecules 2025, 30(3), 446; https://doi.org/10.3390/molecules30030446 - 21 Jan 2025
Viewed by 316
Abstract
Nanoparticle (NP)-mediated gene delivery offers a promising alternative to traditional methods in plant biotechnology, facilitating genetic transformations with enhanced precision and efficiency. This review discusses key factors influencing NP efficacy, including plant cell wall composition, DNA/NP ratios, exposure time, cargo loading, and post-transformation [...] Read more.
Nanoparticle (NP)-mediated gene delivery offers a promising alternative to traditional methods in plant biotechnology, facilitating genetic transformations with enhanced precision and efficiency. This review discusses key factors influencing NP efficacy, including plant cell wall composition, DNA/NP ratios, exposure time, cargo loading, and post-transformation assessments. We explore the challenges of NP cytotoxicity, transformation efficiency, and regeneration while addressing environmental impacts and regulatory considerations. We emphasize the potential for stimulus-responsive NPs and scalable delivery methods to optimize gene editing in agriculture. Full article
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<p>Graphical presentation of different nanomaterials as a suitable substitute for plant gene delivery systems. NPs are selected based on several key properties to optimize their efficacy in biomolecule delivery, such as biocompatibility, encapsulation/binding efficiency, solubility, size, shape, charge, and surface properties. The figure was created with BioRender.com.</p>
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<p>Overview of the key steps in developing a successful protocol for nanoparticle-based plant transformation. The process can be broken down into three major phases: before, during, and after transformation. Before transformation, the intrinsic properties of nanoparticles greatly influence their efficacy. The properties of the plant and the cargo must also be considered. During transformation, the exposure time, buffer conditions, and the amount of cargo loaded into the nanoparticles will be important for the process outcome. After transformation, the plant cells or tissue will need to be propagated, which can have biosafety and environmental consequences. The figure was created with BioRender.com.</p>
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<p>Summary of various delivery methods of nanoparticles and genetic material introduced in the plant cells. The figure was created with BioRender.com.</p>
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<p>Scalability challenges and solutions in NP-mediated gene transformation: from tissue culture bottlenecks to aerosol-mediated foliar spray. The figure was created with BioRender.com.</p>
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20 pages, 4809 KiB  
Article
Design and Evaluation of Noise Simulation Algorithm Using MATLAB Ray Tracing Engine for Noise Assessment and Prediction
by Precin Kalisalvan, Mohd Sayuti Ab Karim and Siti Nurmaya Musa
Appl. Sci. 2025, 15(3), 1009; https://doi.org/10.3390/app15031009 - 21 Jan 2025
Viewed by 297
Abstract
The Malaysian Department of Occupational Safety and Health (DOSH) reported that noise-induced hearing loss (NIHL) accounted for 92% of occupational diseases in 2019. To address this, accurate risk assessment is crucial. The current noise evaluation methods are complex and time-consuming, relying on manual [...] Read more.
The Malaysian Department of Occupational Safety and Health (DOSH) reported that noise-induced hearing loss (NIHL) accounted for 92% of occupational diseases in 2019. To address this, accurate risk assessment is crucial. The current noise evaluation methods are complex and time-consuming, relying on manual calculations and field measurements. An easy-to-use, open-source noise simulator that directly compares the output with national standards would help mitigate this issue. This research aims to develop an advanced noise evaluation tool to assess and predict unregulated workplace noise, providing tailored safety recommendations. Using a representative plant layout, the Sound Pressure Level (SPL) is calculated using MATLAB’s ray tracing propagation model. The model simulates all possible transmission paths from the source to the receiver to derive the resultant SPL. A noise simulation application featuring a graphical user interface (GUI) built with MATLAB’s App Designer (version: R2024a) automates these computations. The simulation results are validated against the DOSH’s safety standards in Malaysia. Additional safety metrics, such as the recommended maximum exposure time and the required Noise Reduction Rating (NRR) for hearing protection, are calculated based on the SPLs for hazardous locations. The simulation algorithm’s functionality is validated against manual calculations, with an average deviation of just 3.06 dB, demonstrating the model’s precision. This tool can assess and predict indoor noise levels, provide information on optimal exposure limits, and recommend necessary protective measures, ultimately reducing the risk of NIHL in factory environments. It can potentially optimise plant floor operations for existing and new facilities, ensuring safer shift operations and reducing worker noise hazard exposure. Full article
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<p>Acoustic ray tracing algorithm framework [<a href="#B31-applsci-15-01009" class="html-bibr">31</a>].</p>
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<p>Overall process of noise assessment and safety recommendations.</p>
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<p>Sections in the graphical user interface (1—file upload, 2—receiver information, 3—source information, 4—safety recommendations, and 5—resultant SPL).</p>
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<p>GUI simulates safety recommendations when the resultant SPL is above the limits.</p>
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<p>Test floor plan with sources and receiver in meters.</p>
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<p>(<b>a</b>) Floor plan of the factory (Furniture Painting and Assembly factory in Klang Malaysia) with pictures (A1/A2—Painting booth, B1/B2—Sanding booth, C1—Assembly booth); (<b>b</b>) drawing of floor plan divided into grids; (<b>c</b>) 3D model of the floor plan’s outline.</p>
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<p>Three-dimensional version of ray diagram of the simulation when reflection is changed from 0, 1, 2, 3 and 4.</p>
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<p>Resultant SPL vs. max number of reflections graph.</p>
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<p>Resultant SPLs vs. surface materials.</p>
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<p>Three-dimensional version of ray diagram during simulation verification (with reflection).</p>
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<p>Recorded SPLs from collected real-world data in dB.</p>
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<p>Simulated SPLs plotted on a grid and heatmap in dB.</p>
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<p>Comparison between SPLs from site and simulation.</p>
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21 pages, 9155 KiB  
Article
Antimicrobial Activity of UV-Activated and Cysteamine-Grafted Polymer Foils Against Bacteria and Algae
by Viktorie Neubertová, Tereza Silovská, Václav Švorčík and Zdeňka Kolská
Polymers 2025, 17(2), 251; https://doi.org/10.3390/polym17020251 - 20 Jan 2025
Viewed by 437
Abstract
Surface modification of various polymer foils was achieved by UV activation and chemical grafting with cysteamine to improve surface properties and antimicrobial efficacy. UVC activation at 254 nm led to changes in surface wettability and charge density, which allowed the introduction of amino [...] Read more.
Surface modification of various polymer foils was achieved by UV activation and chemical grafting with cysteamine to improve surface properties and antimicrobial efficacy. UVC activation at 254 nm led to changes in surface wettability and charge density, which allowed the introduction of amino and thiol functional groups by cysteamine grafting. X-ray photoelectron spectroscopy (XPS) confirmed increased nitrogen and sulfur content on the modified surfaces. SEM analysis revealed that UV activation and cysteamine grafting resulted in distinct surface roughness and texturing, which are expected to enhance microbial interactions. Antimicrobial tests showed increased resistance to algal growth (inhibition test) and bacterial colonization (drop plate method), with significant improvement observed for polyethylene terephthalate (PET) and polyetheretherketone (PEEK) foils. The important factors influencing the efficacy included UV exposure time and cysteamine concentration, with longer exposure and higher concentrations leading to bacterial reduction of up to 45.7% for Escherichia coli and 55.6% for Staphylococcus epidermidis. These findings highlight the potential of combining UV activation and cysteamine grafting as an effective method for developing polymeric materials with enhanced antimicrobial function, offering applications in industries such as healthcare and packaging. Full article
(This article belongs to the Section Polymer Applications)
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<p>Comparison of contact angle values of all pristine and functionalized polymers under study: (<b>a</b>) PET and PEEK; (<b>b</b>) PTFE and PVDF; (<b>c</b>) UPVC and PVDC; (<b>d</b>) PP, UHMWPE, PS, and POM.</p>
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<p>Comparison of zeta potential values of all pristine and functionalized polymers under study: (<b>a</b>) PET and PEEK; (<b>b</b>) PTFE and PVDF; (<b>c</b>) UPVC and PVDC; (<b>d</b>) PP, UHMWPE, PS, and POM.</p>
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<p>Fitted XPS spectra of pristine and modified UPVC samples, showing specific regions of interest. (<b>a</b>) C 1s spectrum of pristine sample, (<b>b</b>) C 1s spectrum of UV60_CYS10, (<b>c</b>) N 1s spectrum of UV60_CYS10, and (<b>d</b>) S 2p spectrum of UV60_CYS10.</p>
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<p>S (2p) sulfur spectrum on UPVC samples. From bottom: pristine (red), UV10_CYS10 (green), UV30_CYS10 (purple), and UV60_CYS10 (azure).</p>
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<p>SEM micrographs of pristine, UV60, and UV60_CYS30 surfaces at 70k× magnification for (<b>a</b>) UPVC, (<b>b</b>) PET, and (<b>c</b>) PEEK. The accelerating voltage was 5 kV, and the scale bar represents 500 nm.</p>
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<p>Results of <span class="html-italic">D. quadricauda</span> algal growth inhibition test for all samples of (<b>a</b>) UPVC and (<b>b</b>) PEEK.</p>
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<p>Numbers of CFUs of <span class="html-italic">E. coli</span> and <span class="html-italic">S. epidermidis</span> for samples of (<b>a</b>) PVDF, (<b>b</b>) UPVC, (<b>c</b>) PET, and (<b>d</b>) PEEK.</p>
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22 pages, 4845 KiB  
Article
Batch Adsorption of Orange II Dye on a New Green Hydrogel—Study on Working Parameters and Process Enhancement
by Andrei-Ionuț Simion, Cristina-Gabriela Grigoraș and Lidia Favier
Gels 2025, 11(1), 79; https://doi.org/10.3390/gels11010079 - 20 Jan 2025
Viewed by 257
Abstract
A new green hydrogel consisting of cherry stone (CS) powder and sodium alginate (SA) was synthesized through physical crosslinking. The product had a mean diameter of 3.95 mm, a moisture content of 92.28%, a bulk density of 0.58 g/cm3, and a [...] Read more.
A new green hydrogel consisting of cherry stone (CS) powder and sodium alginate (SA) was synthesized through physical crosslinking. The product had a mean diameter of 3.95 mm, a moisture content of 92.28%, a bulk density of 0.58 g/cm3, and a swelling ratio of 45.10%. The analyses of its morphological structure and functional groups by scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR) showed the successful entrapping of the CS in the SA polymeric matrix. The viability of the prepared hydrogel as adsorbent was tested towards Orange II (OII) anionic dye. The influence of the pH, adsorbent amount, contact time, and initial dye concentration was evaluated. Then, the impact of three accelerating factors (stirring speed, ultrasound exposure duration, and temperature) on the OII retention was investigated. The highest recorded removal efficiency and adsorption capacity were 82.20% and 6.84 mg/g, respectively. The adsorption followed Elovich and pseudo-second-order kinetics, was adequately described by Freundlich and Khan isotherms, and can be defined as spontaneous, endothermic, and random. The experiments confirmed that the obtained hydrogel can be used acceptably for at least two consecutive cycles, sustaining its effectiveness in water decontamination. Full article
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<p>SEM images of the CSSA hydrogel before (<b>A</b>) and after (<b>B</b>) OII adsorption.</p>
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<p>FTIR spectra of CSSA hydrogel before (<b>A</b>) and after (<b>B</b>) OII adsorption.</p>
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<p><span class="html-italic">p</span>H<sub>PZC</sub> of CSSA hydrogel.</p>
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<p>Influence of <span class="html-italic">p</span>H (<b>A</b>), adsorbent dose (<b>B</b>), time (<b>C</b>), and initial dye concentration (<b>D</b>) on the removal of OII from aqueous solution through adsorption on CSSA hydrogel (the experiments were carried out with 10 mL of OII, at room temperature, without stirring, the working conditions being as follows A: OII concentration—20 mg/L, <span class="html-italic">p</span>H—variable, adsorbent dose—0.038 g/L, contact time—300 min; B: OII concentration—20 mg/L, <span class="html-italic">p</span>H—3, adsorbent dose—variable, contact time—300 min; C: OII concentration—20 mg/L, <span class="html-italic">p</span>H—3, adsorbent dose—0.058 g/L, contact time—variable; D: OII concentration—variable, <span class="html-italic">p</span>H—3, adsorbent dose—0.058 g/L, contact time—300 min).</p>
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<p>Kinetic models of the OII adsorption on CSSA hydrogel (initial OII concentration: (<b>A</b>) 10 mg/L, (<b>B</b>) 15 mg/L, (<b>C</b>) 20 mg/L, (<b>D</b>) 25 mg/L, (<b>E</b>) 30 mg/L, (<b>F</b>) 35 mg/L, (<b>G</b>) 40 mg/L, (<b>H</b>) 45 mg/L, (<b>I</b>) 50 mg/L) (the experiments were carried out with 10 mL of OII, at room temperature, without stirring, the working conditions being as follows: <span class="html-italic">p</span>H—3, adsorbent dose—0.058 g/L, contact time—variable).</p>
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<p>Equilibrium isotherms for the OII adsorption on CSSA hydrogel (the experiments were carried out with 10 mL of OII, at room temperature, without stirring, the working conditions being as follows: OII concentration—variable, <span class="html-italic">p</span>H—3, adsorbent dose—0.058 g/L, contact time—1440 min).</p>
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<p>Influence of stirring speed (<b>A</b>), ultrasound exposure (<b>B</b>), and temperature (<b>C</b>) on the removal of OII from aqueous solution through adsorption on CSSA hydrogel (the experiments were carried out with 10 mL of OII, having <span class="html-italic">p</span>H 3 and a concentration of 20 mg/L, with an adsorbent dose of 0.058 g/L, for 300 min).</p>
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<p>Van ’t Hoff plot for the adsorption of OII on CSSA hydrogel.</p>
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<p>Adsorption–desorption cycles for OII–CSSA hydrogel system (the adsorption experiments were carried out with 10 mL of OII having a concentration of 20 mg/L, and <span class="html-italic">p</span>H 3, at room temperature under stirring at 150 rpm, for 300 min; the desorption experiments were carried out for 240 min, with 50 mL of eluent, under stirring at 150 rpm).</p>
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